2,785 results on '"GLUCOSYLTRANSFERASE"'
Search Results
2. Genome-wide identification of UDP-glycosyltransferases involved in flavonol glycosylation induced by UV-B irradiation in Eriobotrya japonica
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Qian, Jiafei, Ren, Chuanhong, Wang, Fan, Cao, Yunlin, Guo, Yan, Zhao, Xiaoyong, Liu, Yilong, Zhu, Changqing, Li, Xiaoying, Xu, Hongxia, Chen, Junwei, Chen, Kunsong, and Li, Xian
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- 2025
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3. Reversible Glc‐conjugation/hydrolysis modulates the homeostasis of lunularic acid in Marchantia polymorpha growth.
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Zhu, Ting‐Ting, Xu, Yu‐Liang, Ta, He, Zhang, Jiao‐Zhen, Xu, Dan‐Dan, Fu, Jie, Hao, Yue, Du, Ni‐Hong, Cheng, Ai‐Xia, and Lou, Hong‐Xiang
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PLANT growth regulation , *PLANT regulators , *REGULATION of growth , *GROWTH regulators , *GLYCOSYLTRANSFERASES - Abstract
SUMMARY The circadian clock efficiently coordinates growth regulators and plant growth in the temporal regulation of physiological processes. The involvement of the growth‐regulator‐clock in governing the spatio‐temporal regulation of plant growth and development remains unexplained in the nonvascular liverworts. In this study, we aimed to assess the relationship between the putative liverwort growth regulator lunularic acid (LA) levels and the growth variation of Marchantia polymorpha according to the circadian clock. LA level exhibited a similar circadian rhythm as gemmalings' accelerated growth during the light phase, and UV‐B stress experiments implied that the surge in LA levels at the start of the light phase could serve as a circadian rhythm‐based prediction for preempting UV‐B injury, with LA serving as a protective shield against UV‐B irradiation. Notably, Glc‐conjugation/hydrolysis, buffering the anabolism‐catabolism of endogenous LA, was rhythmically regulated. Furthermore, the reversible conversion between LA and LA‐4′‐O‐glucoside (mediated by glucosyltransferases MpUGT744A1 and β‐glucosidases MpBGLU2/3) was characterized both in vitro and in vivo and evidenced to be relevant to diurnal variation in LA level. Interaction between MpUGT744A1 and MpBGLU2 ensures the efficient metabolic flux between LA and LA‐GE. These findings shed light on the regulatory mechanisms that govern LA metabolism and the importance of understanding the temporal aspects of LA for Marchantia's adaptive response to environmental stressors such as UV‐B radiation. [ABSTRACT FROM AUTHOR] more...
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- 2024
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4. Allelic Variation in gtfB – gtfC Region of Natural Variant of Streptococcus mutans Without Biofilm Formation.
- Author
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Kimijima, Manami, Narisawa, Naoki, Nakagawa-Nakamura, Tomoyo, and Senpuku, Hidenobu
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HOMOLOGOUS recombination , *STREPTOCOCCUS mutans , *DENTITION , *GLYCOSYLTRANSFERASES , *DENTAL caries - Abstract
Streptococcus mutans is primarily found in biofilms on tooth surfaces and is associated with the development of dental caries. S. mutans synthesizes water-insoluble glucan (WIG) using sucrose as a substrate, inducing the formation of three-dimensional biofilms. WIG is produced by glucosyltransferases (GTFs) encoded by the tandem and highly homologous gtfB and gtfC genes. Conversely, the homologous recombination of gtfB and gtfC readily happens, producing natural variants without WIG. These WIG− variants are thought to have ecologically pleiotropic functions; however, the molecular basis for their appearance is unclear. This study aimed to determine the sequence of the gtfB–gtfC regions of WIG− variants. We sequenced the gtfB–gtfC regions of 23 WIG− variants derived from S. mutans UA159 and revealed the presence of five alleles and four types of single homologous recombination patterns. Regardless of the allele type, the WIG− variants showed low biofilm formation and GTF activity. To the best of our knowledge, this is the first study to report the presence of alleles in WIG− variants. These findings provide important information for explaining the appearance of mechanisms in WIG− variants. [ABSTRACT FROM AUTHOR] more...
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- 2024
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5. Enzymatic Glycosylation of 4′-Hydroxychalcones: Expanding the Scope of Nature's Catalytic Potential.
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Chlipała, Paweł, Matera, Agata, Sordon, Sandra, Popłoński, Jarosław, Mazur, Marcelina, and Janeczko, Tomasz
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DRUG discovery , *SOYBEAN , *FLAVONOIDS , *CHINESE skullcap , *CHALCONES , *CHALCONE - Abstract
Chalcones, including 4′-hydroxychalcones, have garnered significant attention in the area of drug discovery due to their diverse pharmacological properties, such as anti-inflammatory, antioxidative, and anticancer effects. However, their low water solubility and bioavailability limit their efficacy in vivo. Glycosylation presents a promising approach to enhance the water solubility, stability, and metabolic properties of chalcones. This study investigates the enzymatic glycosylation of eight chemically synthesized 4′-hydroxychalcones using a diverse set of sugar glucosyltransferases from bacterial, plant, and fungal sources, alongside Glycine max sucrose synthase (GmSuSy) in a cascade reaction. Among the tested enzymes, five exhibited a remarkable versatility for glycoside production, and for large-scale biotransformation, flavonoid 7-O-glucosyltransferase Sbaic7OGT from Scutellaria baicalensis was selected as the most effective. As a result of the experiments conducted, eight trans-chalcone glycosides were obtained. During the purification of the reaction products, we also observed the isomerization of the products by simple sunlight exposure, which resulted in eight additional cis-chalcone glycosides. This study highlights the novel use of a cascade reaction involving Glycine max sucrose synthase (GmSuSy) for the efficient glycosylation of trans-4′-hydroxychalcones, alongside the unexpected discovery of cis-chalcone glycosides during the purification process. [ABSTRACT FROM AUTHOR] more...
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- 2024
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6. Monitoring the oral administration of a beneficial microbes mixture based on Apilactobacillus kunkeei strains, in honey bees.
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Arredondo, Daniela, Zunino, Pablo, and Antúnez, Karina
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The administration of beneficial microbes or probiotics is a widely used strategy to improve human and animal health. In a previous study, we developed a mixture of Apilactobacillus kunkeei strains that, when administered to honey bees, reduced the infection by pathogens Nosema ceranae or the mortality associated with Paenibacillus larvae. However, as results obtained by probiotic treatments are variable, it is necessary to monitor its adequate administration. Here, we designed specific primers to detect and quantify A. kunkeei within honey bee guts which can be a helpful strategy to monitor its administration and persistence at the field level in future studies. [ABSTRACT FROM AUTHOR] more...
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- 2024
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7. Allelic Variation in gtfB–gtfC Region of Natural Variant of Streptococcus mutans Without Biofilm Formation
- Author
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Manami Kimijima, Naoki Narisawa, Tomoyo Nakagawa-Nakamura, and Hidenobu Senpuku
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Streptococcus mutans ,homologous recombination ,glucosyltransferase ,water-insoluble glucan ,natural variant ,biofilm ,Specialties of internal medicine ,RC581-951 - Abstract
Streptococcus mutans is primarily found in biofilms on tooth surfaces and is associated with the development of dental caries. S. mutans synthesizes water-insoluble glucan (WIG) using sucrose as a substrate, inducing the formation of three-dimensional biofilms. WIG is produced by glucosyltransferases (GTFs) encoded by the tandem and highly homologous gtfB and gtfC genes. Conversely, the homologous recombination of gtfB and gtfC readily happens, producing natural variants without WIG. These WIG− variants are thought to have ecologically pleiotropic functions; however, the molecular basis for their appearance is unclear. This study aimed to determine the sequence of the gtfB–gtfC regions of WIG− variants. We sequenced the gtfB–gtfC regions of 23 WIG− variants derived from S. mutans UA159 and revealed the presence of five alleles and four types of single homologous recombination patterns. Regardless of the allele type, the WIG− variants showed low biofilm formation and GTF activity. To the best of our knowledge, this is the first study to report the presence of alleles in WIG− variants. These findings provide important information for explaining the appearance of mechanisms in WIG− variants. more...
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- 2024
- Full Text
- View/download PDF
8. Anti-biofilm and anti-glucosyltransferase effects of nano liposomal plant extracts against Streptococcus mutans
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Faraz Radmand, Milad Baseri, Mohammad Yousef Memar, Alireza Ebrahimi, Hamed Hamishehkar, Solmaz Asnaashari, Amirreza Naseri, and Maryam Kouhsoltani
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Nanoliposome ,Streptococcus mutans ,Plant extract ,Biofilm ,Glucosyltransferase ,Medicine ,Science - Abstract
Abstract The role of Streptococcus mutans in the initiation of caries is related to its acidogenicity, aciduricity, and polysaccharides extracellular layer production by glucosyltransferases in dental biofilms. Therefore, inhibition of glucosyltransferase activity impairs the virulence of cariogenic biofilms, which can be used to prevent dental caries. We evaluated the anti-bacterial, anti-biofilm, and anti-glucosyltransferases effects of nanoliposomal herbal aqueous extracts of Liquorice (Glycyrrhiza glabra; G. glabra), Ginger (Zingiber officinale; Z. officinale), Pomegranate (Punica granatum; P. granatum), and Rose (Rosa damascene; R. damascene) via minimum bactericidal concentration and minimum inhibitory concentration against Streptococcus mutans strain ATCC 35,668. An anti-biofilm assay was performed using a minimum biofilm inhibitory concentration test. Among herbs, only P. granatum showed an antibacterial effect. Therefore, a nanoliposomal formulation of P. granatum was developed and characterized. Its effect on S.mutans glucosyltransferases was assessed by measuring glucan amount. The nanoliposomal formulation of P.granatum showed a significantly higher anti-biofilm effect than P. granatum aqueous extract. Their similar potential in blocking glucosyltransferases showed that the nanoliposomal formulation of P.granatum blocked other pathways rather than blocking glucosyltransferases for its anti-biofilm effect. Collectively, the nanoliposomal formulation of P.granatum, due to its anti- Streptococcus mutans characteristics, would be a production which open a new horizon for the oral pharmaceutical industry. more...
- Published
- 2024
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9. Anthocyanin glucosylation mediated by a glycoside hydrolase family 3 protein in purple carrot.
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Koga, Shun‐ya, Miyahara, Taira, Nishizaki, Yuzo, Tamura, Kotaro, Okamoto, Emi, Kawagishi, Hiroaki, Sakurai, Kaori, Kaneko, Yumika, Kumakubo, Ryota, Tanaka, Tsuyoshi, Ozeki, Yoshihiro, and Sasaki, Nobuhiro more...
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AMINO acid sequence , *CYANIDIN , *RECOMBINANT proteins , *NICOTIANA benthamiana , *BIOSYNTHESIS , *ANTHOCYANINS - Abstract
SUMMARY: Purple carrot accumulates anthocyanins modified with galactose, xylose, glucose, and sinapic acid. Most of the genes associated with anthocyanin biosynthesis have been identified, except for the glucosyltransferase genes involved in the step before the acylation in purple carrot. Anthocyanins are commonly glycosylated in reactions catalyzed by UDP‐sugar‐dependent glycosyltransferases (UGTs). Although many studies have been conducted on UGTs, the glucosylation of carrot anthocyanins remains unknown. Acyl‐glucose‐dependent glucosyltransferase activity modifying cyanidin 3‐xylosylgalactoside was detected in the crude protein extract prepared from purple carrot cultured cells. In addition, the corresponding enzyme was purified. The cDNA encoding this glucosyltransferase was isolated based on the partial amino acid sequence of the purified protein. The recombinant protein produced in Nicotiana benthamiana leaves via agroinfiltration exhibited anthocyanin glucosyltransferase activity. This glucosyltransferase belongs to the glycoside hydrolase family 3 (GH3). The expression pattern of the gene encoding this GH3‐type anthocyanin glucosyltransferase was consistent with anthocyanin accumulation in carrot tissues and cultured cells. Significance Statement: Glucosylation of the purple carrot anthocyanin cyanidin 3‐O‐(2″‐O‐xylosyl)‐(6″‐O‐sinapoylglucosyl)‐galactoside is mediated by a glycoside hydrolase family 3 vacuolar enzyme that catalyzes a reaction that uses acyl‐glucose as the glucosyl donor. [ABSTRACT FROM AUTHOR] more...
- Published
- 2024
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10. Anti-biofilm and anti-glucosyltransferase effects of nano liposomal plant extracts against Streptococcus mutans
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Radmand, Faraz, Baseri, Milad, Memar, Mohammad Yousef, Ebrahimi, Alireza, Hamishehkar, Hamed, Asnaashari, Solmaz, Naseri, Amirreza, and Kouhsoltani, Maryam
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- 2024
- Full Text
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11. A new glucosyltransferase UGT78 from Iris sanguinea is a putative negative regulator in cadmium stress response
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Shi, Gongfa, Liu, Guiling, Liu, Huijun, Wang, Lei, Kuwantai, Aerdake, Du, Yu, Wang, Ling, Xi, Xiaolei, and Chai, Rusong
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- 2024
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12. Two UGT84A Family Glycosyltransferases Regulate Phenol, Flavonoid, and Tannin Metabolism in Juglans regia (English Walnut)
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Saxe, Houston J, Horibe, Takanori, Balan, Bipin, Butterfield, Timothy S, Feinberg, Noah G, Zabaneh, Christopher M, Jacobson, Aaron E, and Dandekar, Abhaya M
- Subjects
Agricultural ,Veterinary and Food Sciences ,Plant Biology ,Biological Sciences ,Nutrition ,Good Health and Well Being ,glucosyltransferase ,hydrolysable tannin ,UGT84A ,gallic acid ,walnut ,phenol metabolism ,transcriptomics ,metabolomics ,Crop and pasture production ,Plant biology - Abstract
We showed previously that gallic acid is produced in walnut from 3-dehydroshikimate by a shikimate dehydrogenase (JrSkDH). This study focuses on the next step in the hydrolysable tannin pathway, the formation of 1-O-galloyl-β-D-glucose from the phenolic gallic acid and UDP glucose by a glycosyltransferase. JrGGT1 (UGT84A73) and JrGGT2 (UGT84A74) are predicted to be two such glycosyltransferases, which we expressed in tobacco plants. GC-MS analysis of the transgenic tobacco revealed moderate, yet significant alterations in plant secondary metabolism, such as depleted phenolic acids, including gallic acid. We postulate that these effects are due to JrGGT1 and JrGGT2 activity, as JrGGT orthologs glycosylate these phenolic compounds in vitro. Moreover, JrGGT expression in tobacco caused upregulation of shikimic acid pathway metabolites and differing responses in phenylpropanoids, such as phenolic acids and flavonoids. In transcriptome analysis of walnut pellicle tissues, both JrGGTs showed substantial and significant expression correlations with the gallic acid-producing JrSkDHs and were highly coexpressed with the genetic circuits constituting the shikimic acid and phenylpropanoid biosynthetic pathways. Verification of JrGGT gene expression by transcriptome analysis of 20 walnut tissues revealed striking similarities with that of the pellicle data, with the greatest expression in roots, wood, buds, and leaves of Juglans regia cv. Chandler: tissues that typically accumulate hydrolysable tannins. Like the transgenic tobacco, pellicle metabolomic analyses revealed that many phenylpropanoids correlated negatively with JrGGT expression, while shikimic acid pathway metabolites correlated positively with JrGGT expression. This research supports the hypothesis that JrGGT1 and JrGGT2 play non-trivial roles in metabolism of phenolic acids, flavonoids, and ostensibly, tannins. more...
- Published
- 2021
13. Synergistic inhibitory activity of Glycyrrhizae Radix and Rubi Fructus extracts on biofilm formation of Streptococcus mutans
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Youngseok Ham and Tae-Jong Kim
- Subjects
Biofilm ,Glucosyltransferase ,Glycyrrhizae Radix ,Rubi Fructus ,Streptococcus mutans ,Other systems of medicine ,RZ201-999 - Abstract
Abstract Background Streptococcus mutans is a bacterium that causes oral diseases. Plaque, a biofilm produced by S. mutans and other bacteria, makes it difficult to remove cariogenic oral microorganisms, including biofilm producers. Glucan synthesis by glucosyltransferase is one of the mechanisms underlying plaque formation. This study demonstrates the effectiveness of inhibiting biofilm formation by interfering with the glucosyltransferase activity of S. mutans using edible herbal medicines. Methods This study investigated the inhibitory activity of Glycyrrhizae Radix extract, Rubi Fructus extract, glycyrrhizin from Glycyrrhizae Radix, and ellagic acid from Rubi Fructus against glucosyltransferase activity of S. mutans. Enzyme kinetic analysis identified the mechanism by which glycyrrhizin and ellagic acid inhibit enzyme activity. Results The conditions for synergistically inhibiting biofilm formation by combining Glycyrrhizae Radix and Rubi Fructus extracts were identified. Biofilm formation was also synergistically inhibited by mixing their respective active constituents, glycyrrhizin and ellagic acid. Glycyrrhizin and ellagic acid inhibited glucosyltransferase via noncompetitive and uncompetitive mechanisms, respectively, indicating that they inhibit it via distinct mechanisms. Conclusions This study presents an effective oral hygiene method using the synergistic activity of two natural plant extracts to inhibit biofilm formation through different inhibitory mechanisms against glucosyltransferase of S. mutans. more...
- Published
- 2023
- Full Text
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14. The findings of glucosyltransferase enzymes derived from oral streptococci
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Tomonori Hoshino and Taku Fujiwara
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Glucosyltransferase ,Streptococcus ,Anti-caries vaccine ,Phylogenetic analysis ,Dental caries ,Plaque biofilm (6 key words) ,Dentistry ,RK1-715 - Abstract
Glucosyltransferase enzymes (Gtfs) distribute among some streptococcal species in oral cavity and are known as key enzymes contributing to the development of oral biofilm such as dental plaque. In 18 streptococcal species, 45 glucosyltransferase genes (gtf) are detected from genome database. Gtfs catalyze the synthesis of the glucans, which are polymers of glucose, from sucrose and they are main component of oral biofilm. Especially, the Gtfs from Streptococcus mutans are recognized as one of dental caries pathogens since they contribute to the formation of dental plaque and the establishment of S. mutans in the tooth surface. Therefore, Gtfs has been studied particularly by many researchers in the dentistry field to develop the anti- caries vaccine. However, it is not still accomplished. In these days, the phylogenetic and crystal structure analyses of Gtfs were performed and the study of Gtfs will enter new situation from the technique in the past old viewpoint. The findings from those analyses will affect the development of the anti-caries vaccine very much after this. In this review, we summarize the findings of oral streptococcal Gtfs and consider the perspectives of the dental caries prevention which targeted Gtf. more...
- Published
- 2022
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15. Molecular Identification of UDP-Sugar-Dependent Glycosyltransferase and Acyltransferase Involved in the Phenylethanoid Glycoside Biosynthesis Induced by Methyl Jasmonate in Sesamum indicum L.
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Fuji, Yushiro, Uchida, Kai, Akashi, Tomoyoshi, Ohtsuki, Takashi, Matsufuji, Hiroshi, and Hirai, Masami Yokota
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SESAME , *BIOSYNTHESIS , *ACYLTRANSFERASES , *JASMONATE , *RECOMBINANT proteins , *CELL culture - Abstract
Sesame (Sesamum indicum L.) plants contain large amounts of acteoside, a typical phenylethanoid glycoside (PhG) that exhibits various pharmacological activities. Although there is increasing interest in the biosynthesis of PhGs for improved production, the pathway remains to be clarified. In this study, we established sesame-cultured cells and performed transcriptome analysis of methyl jasmonate (MeJA)–treated cultured cells to identify enzyme genes responsible for glucosylation and acylation in acteoside biosynthesis. Among the genes annotated as UDP-sugar-dependent glycosyltransferase (UGT) and acyltransferase (AT), 34 genes and one gene, respectively, were upregulated by MeJA in accordance with acteoside accumulation. Based on a phylogenetic analysis, five UGT genes (SiUGT1–5) and one AT gene (SiAT1) were selected as candidate genes involved in acteoside biosynthesis. Additionally, two AT genes (SiAT2–3) were selected based on sequence identity. Enzyme assays using recombinant SiUGT proteins revealed that SiUGT1, namely, UGT85AF10, had the highest glucosyltransferase activity among the five candidates against hydroxytyrosol to produce hydroxytyrosol 1- O -glucoside. SiUGT1 also exhibited glucosyltransferase activity against tyrosol to produce salidroside (tyrosol 1- O -glucoside). SiUGT2, namely, UGT85AF11, had similar activity against hydroxytyrosol and tyrosol. Enzyme assay using the recombinant SiATs indicated that SiAT1 and SiAT2 had activity transferring the caffeoyl group to hydroxytyrosol 1- O -glucoside and salidroside (tyrosol 1- O -glucoside) but not to decaffeoyl-acteoside. The caffeoyl group was attached mainly at the 4-position of glucose of hydroxytyrosol 1- O -glucoside, followed by attachment at the 6-position and the 3-position of glucose. Based on our results, we propose an acteoside biosynthetic pathway induced by MeJA treatment in sesame. [ABSTRACT FROM AUTHOR] more...
- Published
- 2023
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16. Functional identification of anthocyanin glucosyltransferase genes: a Ps3GT catalyzes pelargonidin to pelargonidin 3-O-glucoside painting the vivid red flower color of Paeonia.
- Author
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Wang, Qianyu, Zhu, Jin, Li, Bing, Li, Shanshan, Yang, Yong, Wang, Qingyun, Xu, Wenzhong, and Wang, Liangsheng
- Abstract
Main conclusion: Glycosylation from an anthocyanidin 3-O-glucosyltransferase Ps3GT (PsUGT78A27) facilitates the accumulation of pelargonidin 3-O-glucoside, which defines the vivid red flower color and occurs only in specific peony tree cultivars. Although tree peony cultivars of Chinese and Japanese both originated from China, vivid red color is only found in flowers of Japanese cultivars but not of Chinese cultivar groups. In this study, a Japanese tree peony cultivar ‘Taiyoh’ with vivid red petals and a Chinese tree peony cultivar ‘Hu Hong’ with reddish pink petals were chosen as the experimental materials. Flavonoids profiling indicated that pelargonidin 3-O-glucoside (Pg3G) detected only in Japanese cultivar contributed to vivid red color of tree peony petals, while pelargonidin 3,5-di-O-glucoside (Pg3G5G) found in both of Japanese and Chinese cultivars was responsible for pink flower color. Through the integration of full-length transcriptome sequencing and in vitro enzymatic activity analysis, two anthocyanin glucosyltransferase genes PsUGT78A27 and PsUGT75L45 were isolated from the petals of tree peony, and their encoding products exhibited enzymatic activities of pelargonidin 3-O-glucosyltransferase and anthocyanin 5-O-glucosyltransferase, respectively. Further quantitative real-time PCR revealed that PsUGT78A27 displayed high expression in petals of both cultivars and PsUGT75L45 was expressed at high levels in cultivar ‘Hu Hong’ only. Using a gene gun technique, the GFP fusion proteins of PsUGT78A27 and PsUGT75L45 were visualized to be cytoplasmic and nuclear localization in the epidermal cells of tree peony petals, and the glucosylation function of PsUGT78A27 and PsUGT75L45 to alter petal color of tree peony and herbaceous peony had been directly validated in vivo. These results demonstrated that PsUGT78A27 and PsUGT75L45 are key players for the presence or absence of vivid red flower color in tree peony cultivars. Our findings further elucidated the chemical and molecular mechanism of petal pigmentation of Paeonia and could help breed the Paeonia cultivars possessing novel flower colors. [ABSTRACT FROM AUTHOR] more...
- Published
- 2023
- Full Text
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17. Isolation, characterization, and comparative genomic analysis of vB_BviS-A10Y, a novel bacteriophage from mangrove sediments.
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Zhang, Tianyou, Zhang, Menghui, Chen, Yuan, Yu, Meishun, Zeng, Runying, and Jin, Min
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GENOMICS , *BACTERIOPHAGES , *CARBON cycle , *MANGROVE plants , *WHOLE genome sequencing , *VIRAL genomes - Abstract
Mangrove is among the most carbon-rich biomes on earth, and viruses are believed to play a significant role in modulating local and global carbon cycling. However, few viruses have been isolated from mangrove sediments to date. Here, we report the isolation of a novel Bacillus phage (named phage vB_BviS-A10Y) from mangrove sediments. Phage vB_BviS-A10Y has a hexameric head with a diameter of ~ 79.22 nm and a tail with a length of ~ 548.56 nm, which are typical features of siphophages. vB_BviS-A10Y initiated host lysis at 3.5 h postinfection with a burst size of 25 plaque-forming units (PFU)/cell. The genome of phage vB_BviS-A10Y is 162,435 bp long with 225 predicted genes, and the GC content is 34.03%. A comparison of the whole genome sequence of phage vB_BviS-A10Y with those of other phages from the NCBI viral genome database showed that phage vB_BviS-A10Y has the highest similarity (73.7% identity with 33% coverage) to Bacillus phage PBC2. Interestingly, abundant auxiliary metabolic genes (AMGs) were identified in the vB_BviS-A10Y genome. The presence of a β-1,3-glucosyltransferase gene in the phage genome supported our previous hypothesis that mangrove viruses may manipulate carbon cycling directly through their encoded carbohydrate-active enzyme (CAZyme) genes. Therefore, our study will contribute to a better understanding of the diversity and potential roles of viruses in mangrove ecosystems. [ABSTRACT FROM AUTHOR] more...
- Published
- 2023
- Full Text
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18. Synergistic inhibitory activity of Glycyrrhizae Radix and Rubi Fructus extracts on biofilm formation of Streptococcus mutans.
- Author
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Ham, Youngseok and Kim, Tae-Jong
- Subjects
GLYCYRRHIZA ,HIGH performance liquid chromatography ,CONFIDENCE intervals ,BIOFILMS ,PHYTOCHEMICALS ,DYNAMICS ,T-test (Statistics) ,DRUG synergism ,STREPTOCOCCUS mutans ,DESCRIPTIVE statistics ,RESEARCH funding ,PLANT extracts ,BIOLOGICAL assay ,DATA analysis software ,ENZYME inhibitors - Abstract
Background: Streptococcus mutans is a bacterium that causes oral diseases. Plaque, a biofilm produced by S. mutans and other bacteria, makes it difficult to remove cariogenic oral microorganisms, including biofilm producers. Glucan synthesis by glucosyltransferase is one of the mechanisms underlying plaque formation. This study demonstrates the effectiveness of inhibiting biofilm formation by interfering with the glucosyltransferase activity of S. mutans using edible herbal medicines. Methods: This study investigated the inhibitory activity of Glycyrrhizae Radix extract, Rubi Fructus extract, glycyrrhizin from Glycyrrhizae Radix, and ellagic acid from Rubi Fructus against glucosyltransferase activity of S. mutans. Enzyme kinetic analysis identified the mechanism by which glycyrrhizin and ellagic acid inhibit enzyme activity. Results: The conditions for synergistically inhibiting biofilm formation by combining Glycyrrhizae Radix and Rubi Fructus extracts were identified. Biofilm formation was also synergistically inhibited by mixing their respective active constituents, glycyrrhizin and ellagic acid. Glycyrrhizin and ellagic acid inhibited glucosyltransferase via noncompetitive and uncompetitive mechanisms, respectively, indicating that they inhibit it via distinct mechanisms. Conclusions: This study presents an effective oral hygiene method using the synergistic activity of two natural plant extracts to inhibit biofilm formation through different inhibitory mechanisms against glucosyltransferase of S. mutans. [ABSTRACT FROM AUTHOR] more...
- Published
- 2023
- Full Text
- View/download PDF
19. Genome analysis of novel Apilactobacillus sp. isolate from butterfly (Pieris canidia) gut reveals occurrence of unique glucanogenic traits and probiotic potential.
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Jawad, Iqra, Khaliq, Shazia, Akhtar, Nasrin, Saleem, Muhammad, Akhtar, Kalsoom, Ghauri, Komal, and Anwar, Munir A
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MICROBIAL exopolysaccharides , *LACTIC acid , *PROBIOTICS , *NUCLEIC acid hybridization , *LACTIC acid bacteria , *WHOLE genome sequencing , *GENOMES , *BUTTERFLIES - Abstract
This study was conducted with a perception that fructose-rich niches may inhabit novel species of lactic acid bacteria that are gaining importance as probiotics and for the production of exopolysaccharides that have applications in food and pharmaceuticals. Recently, some Lactobacillus species have been reclassified as fructophilic lactic acid bacteria due to their preference for fructose over glucose as a carbon source. These bacteria are likely to be found in fructose rich niches such as flower nectar and insects that feed on it. We explored the butterfly gut and acquired a new isolate, designated as F1, of fructophilic lactic acid bacteria, which produces a glucan-type exopolysaccharide. Whole genome sequencing and in silico analysis revealed that F1 has significantly lower average nucleotide identity and DNA-DNA hybridization values as compared to its closest Apilactobacillus neighbors in phylogenetic analysis. Therefore, we declare the isolate F1 as a novel Apilactobacillus species with the proposed name of Apilactobacillus iqraium F1. Genome mining further revealed that F1 harbors genes for exopolysaccharide synthesis and health-promoting attributes. To this end, F1 is the only Apilactobacillus species harboring three diverse α-glucan-synthesis genes that cluster with different types of dextransucrases in the dendrogram. Moreover, many nutritional marker genes, as well as genes for epithelial cell adhesion and antimicrobial synthesis, were also detected suggesting the probiotic attributes of F1. Overall analysis suggests A. iqraium sp. F1 be a potential candidate for various health beneficial and pharmaceutical applications. [ABSTRACT FROM AUTHOR] more...
- Published
- 2023
- Full Text
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20. Inhibitory effects of Streptococcus salivarius K12 on formation of cariogenic biofilm.
- Author
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Kim, Hye-Ji and Yoo, Hyun-Jun
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BIOFILMS ,STREPTOCOCCUS ,STREPTOCOCCUS mutans ,BACTERIAL DNA - Abstract
Streptococcus salivarius (S. salivarius) K12 is known to be a probiotic bacterium. The purpose of this study was to investigate anti-cariogenic effects of S. salivarius K12 on cariogenic biofilm. S. salivarius K12 was cultured in M17 broth. The antimicrobial activity of spent culture medium (SCM) against Streptococcus mutans was investigated. S. salivarius K12 was co-cultivated with S. mutans using a membrane insert. When the biofilm was formed using salivary bacteria and S. mutans , the K12 was inoculated every day. The biomass of biofilm was investigated by a confocal laser scanning microscope. Also, bacterial DNA from the biofilm was extracted, and then bacteria proportion was analyzed by quantitative PCR using specific primers. The expression of gtf genes of S. mutans in the biofilm with or without S. salivarius K12 was analyzed by RT-PCR. The SCM of S. salivarius K12 inhibited the growth of S. mutans. Also, S. salivarius K12 reduced S. mutans growth in co-cultivation. The formation of cariogenic biofilm was reduced by adding S. salivarius K12, and the count of S. mutans in the biofilm was also decreased in the presence of S. salivarius K12. gtf B, gtf C, and gtf D expression of S. mutans in the biofilm was reduced in the presence of S. salivarius K12. S. salivarius K12 may inhibit the formation of cariogenic biofilm by interrupting the growth and glucosyltransferase production of S. mutans. [ABSTRACT FROM AUTHOR] more...
- Published
- 2023
- Full Text
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21. Evaluation of double expression system for co-expression and co-immobilization of flavonoid glucosylation cascade.
- Author
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Matera, Agata, Dulak, Kinga, Sordon, Sandra, Waśniewski, Kacper, Huszcza, Ewa, and Popłoński, Jarosław
- Subjects
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SOYBEAN , *BACILLUS licheniformis , *GLYCOCONJUGATES , *NATURAL products , *SUCROSE , *FLAVONOIDS - Abstract
Glucosylation cascade consisting of Leloir glycosyltransferase and sucrose synthase with in situ regeneration system of expensive and low available nucleotide sugars is a game-changing strategy for enzyme-based production of glycoconjugates of relevant natural products. We designed a stepwise approach including co-expression and one-step purification and co-immobilization on glass-based EziG resins of sucrose synthase from Glycine max (GmSuSy) with promiscuous glucosyltransferase YjiC from Bacillus licheniformis to produce efficient, robust, and versatile biocatalyst suited for preparative scale flavonoid glucosylation. The undertaken investigations identified optimal reaction conditions (30 °C, pH 7.5, and 10 mM Mg2+) and the best-suited carrier (EziG Opal). The prepared catalyst exhibited excellent reusability, retaining up to 96% of initial activity after 12 cycles of reactions. The semi-preparative glucosylation of poorly soluble isoflavone Biochanin A resulted in the production of 73 mg Sissotrin (Biochanin A 7-O-glucoside). Additionally, the evaluation of the designed double-controlled, monocistronic expression system with two independently induced promoters (rhaBAD and trc) brought beneficial information for dual-expression plasmid design. Key points: • Simultaneous and titratable expression from two independent promoters is possible, although full control over the expression is limited. • Designed catalyst managed to glucosylate poorly soluble isoflavone. • The STY of Sissotrin using the designed catalyst reached 0.26 g/L∙h∙g of the resin. [ABSTRACT FROM AUTHOR] more...
- Published
- 2022
- Full Text
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22. The findings of glucosyltransferase enzymes derived from oral streptococci.
- Author
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Hoshino, Tomonori and Fujiwara, Taku
- Subjects
CARIOGENIC agents ,CAVITY prevention ,STREPTOCOCCUS ,STREPTOCOCCUS mutans ,DENTAL plaque ,ENZYMES - Abstract
Glucosyltransferase enzymes (Gtfs) distribute among some streptococcal species in oral cavity and are known as key enzymes contributing to the development of oral biofilm such as dental plaque. In 18 streptococcal species, 45 glucosyltransferase genes (gtf) are detected from genome database. Gtfs catalyze the synthesis of the glucans, which are polymers of glucose, from sucrose and they are main component of oral biofilm. Especially, the Gtfs from Streptococcus mutans are recognized as one of dental caries pathogens since they contribute to the formation of dental plaque and the establishment of S. mutans in the tooth surface. Therefore, Gtfs has been studied particularly by many researchers in the dentistry field to develop the anti- caries vaccine. However, it is not still accomplished. In these days, the phylogenetic and crystal structure analyses of Gtfs were performed and the study of Gtfs will enter new situation from the technique in the past old viewpoint. The findings from those analyses will affect the development of the anti-caries vaccine very much after this. In this review, we summarize the findings of oral streptococcal Gtfs and consider the perspectives of the dental caries prevention which targeted Gtf. [ABSTRACT FROM AUTHOR] more...
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- 2022
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23. Glucosyltransferase‐modulated Streptococcus mutans adhesion to different surfaces involved in biofilm formation by atomic force microscopy.
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Wang, Rui, Wang, Yigan, Lei, Zixue, Hao, Liying, and Jiang, Li
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BACTERIAL adhesion ,STREPTOCOCCUS mutans ,ATOMIC force microscopy ,BACTERIAL cell surfaces ,BIOFILMS ,DENTAL materials ,SCANNING electron microscopy - Abstract
Biofilm on dental restorative materials is an important determinant in the etiology of secondary caries development. Formation of biofilm involves adhesion of bacteria onto substrate, bacterial cell, and biofilm surfaces. Glucosyltransferase B and C (GtfB and GtfC) are essential factors for regulation of Streptococcus mutans biofilm formation, but the mechanisms involving different kinds of bacterial adhesion still lack detailed description. In this study, nanoscale adhesion force measurement was performed using atomic force microscopy. Bacteria‐coated cantilevers were used to probe S. mutans adhesion to substrates, bacterial cells, and early biofilms. Two representative dental materials, glass ionomer cement (GIC) and composite resin, served as substrates. It was found that deletion of gtfB and gtfC genes both reduced adhesion forces of S. mutans toward substrate and bacterial cell surfaces (P < 0.05). Notably, reduction of the gtfB gene remarkably decreased bacterial adhesion to biofilm surfaces (P < 0.05), while gtfC showed no obvious effect during this stage. Biofilms cultured on GIG further decreased cell–biofilm adhesion, compared with those on resin (P < 0.05). Confocal fluorescence images and scanning electron microscopy images showed that deletion of gtfB lead to reduced microcolony formation and less production of exopolysaccharides (EPSs) in the biofilm, and after bacterial culturing on GIC, the EPS content was further decreased. Our findings suggest that EPSs mainly mediate bacterial adhesion to early biofilm surface. Deletion of gtfB and coculture with GIC could significantly reduce the cell–biofilm adhesion, which is probably through decreasing of EPS production. gtfB exerts a critical role in the bacterial adhesion for the whole process of biofilm development, while gtfC possibly works only in the early stages. [ABSTRACT FROM AUTHOR] more...
- Published
- 2022
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24. Investigating non-target site resistance to pyroxsulam in a glyphosate-resistant Lolium rigidum population.
- Author
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Zeng Y, Liu M, Qian H, Zhao H, Fang Y, Yu Q, Bai L, and Pan L
- Abstract
Background: Resistance to multiple herbicides is common in Lolium rigidum. Here, resistance to acetolactate synthase (ALS)- and susceptibility to acetyl-CoA carboxylase (ACCase)-inhibiting herbicides was confirmed in a glyphosate-resistant L. rigidum population (NLR70) from Australia and the mechanisms of pyroxsulam resistance were examined., Results: No ALS target-site mutations nor gene overexpression were detected. Cytochrome P450 monooxygenase (P450) and glutathione S-transferase (GST) inhibitors (indicators of some certain P450s or GSTs) did not significantly affect the resistance to pyroxsulam. Nevertheless, HPLC analysis showed that plants of the NLR70 population metabolized pyroxsulam faster than plants of the herbicide-susceptible population (SVLR1). RNA sequencing analysis and RT-qPCR validation confirmed that four P450s (CYP709B2, CYP72A14, CYP89A2, CYP94B3), one GT (UGT79), and one ABC transporter (ABCG41) genes were constitutively upregulated in NLR70 plants., Conclusion: This study demonstrates that the glyphosate-resistant L. rigidum population (NLR70) also exhibits resistance to pyroxsulam and identifies six candidate genes associated with non-target site resistance to pyroxsulam. © 2025 Society of Chemical Industry., (© 2025 Society of Chemical Industry.) more...
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- 2025
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25. Reversible Glc-conjugation/hydrolysis modulates the homeostasis of lunularic acid in Marchantia polymorpha growth.
- Author
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Zhu TT, Xu YL, Ta H, Zhang JZ, Xu DD, Fu J, Hao Y, Du NH, Cheng AX, and Lou HX
- Subjects
- Circadian Rhythm physiology, Circadian Clocks, Ultraviolet Rays, Plant Proteins metabolism, Plant Proteins genetics, Gene Expression Regulation, Plant, Glucosyltransferases metabolism, Plant Growth Regulators metabolism, Marchantia metabolism, Marchantia genetics, Marchantia radiation effects, Marchantia growth & development, Marchantia physiology, Homeostasis
- Abstract
The circadian clock efficiently coordinates growth regulators and plant growth in the temporal regulation of physiological processes. The involvement of the growth-regulator-clock in governing the spatio-temporal regulation of plant growth and development remains unexplained in the nonvascular liverworts. In this study, we aimed to assess the relationship between the putative liverwort growth regulator lunularic acid (LA) levels and the growth variation of Marchantia polymorpha according to the circadian clock. LA level exhibited a similar circadian rhythm as gemmalings' accelerated growth during the light phase, and UV-B stress experiments implied that the surge in LA levels at the start of the light phase could serve as a circadian rhythm-based prediction for preempting UV-B injury, with LA serving as a protective shield against UV-B irradiation. Notably, Glc-conjugation/hydrolysis, buffering the anabolism-catabolism of endogenous LA, was rhythmically regulated. Furthermore, the reversible conversion between LA and LA-4'-O-glucoside (mediated by glucosyltransferases MpUGT744A1 and β-glucosidases MpBGLU2/3) was characterized both in vitro and in vivo and evidenced to be relevant to diurnal variation in LA level. Interaction between MpUGT744A1 and MpBGLU2 ensures the efficient metabolic flux between LA and LA-GE. These findings shed light on the regulatory mechanisms that govern LA metabolism and the importance of understanding the temporal aspects of LA for Marchantia's adaptive response to environmental stressors such as UV-B radiation., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.) more...
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- 2025
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26. Novel structural insights for a pair of monoclonal antibodies recognizing non-overlapping epitopes of the glucosyltransferase domain of Clostridium difficile toxin B
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Jinyu Liu, Michael Kothe, Jianxin Zhang, Eliud Oloo, Svetlana Stegalkina, Sophia T. Mundle, Lu Li, Jinrong Zhang, Leah E. Cole, Lucianna Barone, Hans-Peter Biemann, Harry Kleanthous, Natalie G. Anosova, and Stephen F. Anderson more...
- Subjects
Clostridium difficile ,Glucosyltransferase ,Crystal structure ,Antibody ,Protein conformation ,Protein crystallization ,Biology (General) ,QH301-705.5 - Abstract
Clostridium difficile toxins are the primary causative agents for hospital-acquired diarrhea and pseudomembranous colitis. Numerous monoclonal antibodies (mAbs) targeting different domains of Clostridium difficile toxin have been reported. Here we report the crystal structures of two mAbs, B1 and B2, in complex with the glycosyltransferase domain (GTD) of the Clostridium difficile toxin B (TcdB). B2 bound to the N-terminal 4 helix bundle of the GTD, a conserved membrane localization domain (MLD) found in the large clostridial glycosylating toxin family implicated in targeting plasma membrane. B1 bound to a distinct epitope at the hinge region between the MLD and the catalytic subdomain of the GTD. Functional studies revealed the potency of these mAbs in vitro and in vivo to be synergistic when given in combination. more...
- Published
- 2022
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27. Anti-Biofilm Activities of Chinese Poplar Propolis Essential Oil against Streptococcus mutans.
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Yuan, Jie, Yuan, Wenqin, Guo, Yuyang, Wu, Qian, Wang, Fei, and Xuan, Hongzhuan
- Abstract
Streptococcus mutans (S. mutans) is a common cariogenic bacterium that secretes glucosyltransferases (GTFs) to synthesize extracellular polysaccharides (EPSs) and plays an important role in plaque formation. Propolis essential oil (PEO) is one of the main components of propolis, and its antibacterial activity has been proven. However, little is known about the potential effects of PEO against S. mutans. We found that PEO has antibacterial effects against S. mutans by decreasing bacterial viability within the biofilm, as demonstrated by the XTT assay, live/dead staining assay, LDH activity assay, and leakage of calcium ions. Furthermore, PEO also suppresses the total of biofilm biomasses and damages the biofilm structure. The underlying mechanisms involved may be related to inhibiting bacterial adhesion and GTFs activity, resulting in decreased production of EPSs. In addition, a CCK8 assay suggests that PEO has no cytotoxicity on normal oral epithelial cells. Overall, PEO has great potential for preventing and treating oral bacterial infections caused by S. mutans. [ABSTRACT FROM AUTHOR] more...
- Published
- 2022
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28. Chemoenzymatic synthesis of an α-1,6-glucan-based conjugate vaccine against Helicobacter pylori.
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Altman, Eleonora, Chandan, Vandana, Harrison, Blair A, Schur, Melissa, Goneau, Marie-France, Li, Jianjun, and Gilbert, Michel
- Subjects
- *
HELICOBACTER pylori , *CLARITHROMYCIN , *GLUCANS , *TETANUS vaccines , *SERUM albumin , *GLYCOCONJUGATES , *VACCINES - Abstract
In this study, we investigated the utility of glycoconjugates based on a linear α-1,6-glucan chain synthesized using a recombinant α-1,6-glucosyltransferase from the 26695 strain of Helicobacter pylori. Capillary electrophoresis–mass spectrometry analysis confirmed the main product to contain 9–10 sequentially added α-1,6-linked glucose residues. This was consistent with a length of α-1,6-glucan structure present in the outer core region of H. pylori lipopolysaccharide (LPS) from strains 26695 and 26695 HP0826::Kan. The synthetic α-1,6-glucan was conjugated to either bovine serum albumin or tetanus toxoid and immunological properties of resultant glycoconjugates investigated. The conjugates were immunogenic in rabbits and mice and induced strong and specific IgG responses against purified LPS from typeable and nontypeable α-1,6-glucan-positive H. pylori strains. Furthermore, the post-immune sera from rabbits that received the conjugates were bactericidal and cross-reacted with selected clarithromycin-resistant and clarithromycin-susceptible clinical isolates of H. pylori. This technology offers a novel approach to the design of a synthetic carbohydrate-based vaccine against H. pylori. [ABSTRACT FROM AUTHOR] more...
- Published
- 2022
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29. Maltogenic amylase and glucosyltransferase synergistically control the chain structure of rice starch and their effects on rheological properties and functional characteristics of rice milk.
- Author
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Xu, Xiankang, Peng, Xinyun, Zheng, Bo, Zhu, Junchao, and Chen, Ling
- Subjects
- *
SHORT-chain fatty acids , *DAIRY products , *GLYCEMIC index , *RHEOLOGY , *WATER storage , *RICE starch , *RICE products - Abstract
Maltogenic amylase (MA) and glucosyltransferase (TG) were used synergistically for the development of rice milk products with low glycemic index (GI), and its storage properties and nutritional functions were evaluated. The structural results showed that MA and TG treatment could effectively reduce the molecular weight of rice starch and increases its overall branching degree, which further affected the storage and nutritional properties of rice milk products. Compared with gelatinized rice milk and rice milk treated with MA or TG, MTRM (treated with MA and TG) had lower paste viscosity, stable storage modulus, and stronger ability to bind water during storage period, indicating that the dual-enzyme treatment significantly improved the retrogradation properties of rice milk and the anti-retrogradation rate reached 76.12%. In vitro studies showed that dual-enzyme treatment significantly increased the resistant starch content of rice milk to 39.46%, thereby reducing its predicted glycemic index to 53.85, improving its antioxidant properties and contributing to the proliferation of short-chain fatty acids produced bacteria. Notably, MTRM significantly elevated butyrate content which showed a better prebiotic effect. This research provides new approaches for the preparation of low-GI rice-based beverages. [Display omitted] • MA and TG increased the overall branching degree of rice starch. • Low-GI rice milk with stronger antioxidant capacity was prepared by MA and TG. • Paste viscosity and retrogradation rate of MTRM was significantly decreased. • MTRM showed stable storage modulus and stronger ability to bind water during storage. • MTRM contribute to the proliferation of SCFA-produced bacteria to produce SCFAs. [ABSTRACT FROM AUTHOR] more...
- Published
- 2025
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30. High-level de novo biosynthesis of glycosylated zeaxanthin and astaxanthin in Escherichia coli
- Author
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Xixian Chen, Xiaohui Lim, Aurélie Bouin, Thomas Lautier, and Congqiang Zhang
- Subjects
Carotenoids ,Metabolic engineering ,Synthetic biology ,Glycosylation ,Glucosyltransferase ,Zeaxanthin ,Technology ,Chemical technology ,TP1-1185 ,Biotechnology ,TP248.13-248.65 - Abstract
Abstract Because of wide applications in food, feed, pharmaceutical and cosmetic industries, the carotenoid market is growing rapidly. Most carotenoids are hydrophobic, which limits their bioavailability. Glycosylation is a natural route that substantially increases the water solubility, as well as the bioavailability, photostability and biological activities of carotenoids. Here, we report metabolic engineering efforts (e.g., promoter and RBS engineering, optimization of carbon sources and supplementation of bottleneck genes) to produce glycosylated carotenoids in Escherichia coli. By fine-tuning the carotenoid-biosynthetic genes (crtX, crtZ and crtY), our strain produced up to 47.2 mg/L (~ 11,670 ppm) of zeaxanthin glucosides, ~ 78% of the total carotenoids produced. In another construct with mevalonate, astaxanthin pathway and crtX genes, the strain produced a mixture of carotenoid glucosides including astaxanthin and adonixanthin glucosides with a total yield of 8.1 mg/L (1774 ppm). Our work demonstrated a proof-of-concept study for the microbial biosynthesis of glycosylated carotenoids. more...
- Published
- 2021
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31. Tolerance of Arabidopsis thaliana plants overexpressing grapevine VaSTS1 or VaSTS7 genes to cold, heat, drought, salinity, and ultraviolet irradiation
- Author
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Z.V. OGNEVA, O.A. ALEYNOVA, A.R. SUPRUN, Y.A. KARETIN, A.S. DUBROVINA, and K.V. KISELEV
- Subjects
glucosyltransferase ,peroxidase ,piceid ,resveratrol ,stilbene synthase ,viniferin ,Biology (General) ,QH301-705.5 ,Plant ecology ,QK900-989 - Abstract
Stilbene synthases (STS) are plant enzymes that are responsible for the biosynthesis of stilbenes, which are plant phenolic compounds with valuable biological properties. Stilbenes also play important roles in plant tolerance to biotic and abiotic stresses. Therefore, plants that overexpress STS genes can be more resistant to various stresses. This paper investigated the effects of STS gene overexpression in Arabidopsis thaliana (L.) Heynh. Columbia-0 plants on stilbene content and tolerance to the following abiotic stresses: low and high temperatures, salinity, drought, and ultraviolet irradiation (UV-B and UV-C). We used VaSTS1 and VaSTS7 genes from grapevine (Vitis amurensis Rupr.) expressed under the double cauliflower mosaic virus 35S (CaMV35S) promoter. This study firstly demonstrated that overexpression of the VaSTS1 and VaSTS7 genes in A. thaliana plants considerably increased plant tolerance to UV-B and UV-C, while the tolerance to the low and high temperatures, salinity, and drought was not affected. We showed that the highest trans-piceid and trans-resveratrol total content was in ST1 A. thaliana plants that overexpressed the VaSTS1 gene in the range 8.28 - 22.66 µg g-1(f.m.). ST7 plants that overexpressed the VaSTS7 gene showed only trans-resveratrol at 0.02 - 0.08 µg g-1(f.m). Stilbene content and UV tolerance in transgenic A. thaliana plants correlated with STS transgene expression. STS expression, UV tolerance, and stilbene content was higher in VaSTS1 transgenic plants compared with that in VaSTS7 transgenic plants. more...
- Published
- 2021
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32. Targeting glucosyltransferases to combat dental caries: Current perspectives and future prospects.
- Author
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Atta, Lubna, Mushtaq, Mamona, Siddiqui, Ali Raza, Khalid, Assad, and Ul-Haq, Zaheer
- Subjects
- *
ORAL microbiology , *DRUG resistance in bacteria , *DENTAL caries , *STREPTOCOCCUS mutans , *DRUG resistance in microorganisms , *BIOFILMS - Abstract
The emergence of antimicrobial resistance within bacterial communities poses formidable challenges to existing therapeutic strategies aimed at mitigating biofilm-mediated infections. Recent advancements in this domain have spurred the development of targeted antimicrobial agents, designed to selectively eradicate the primary etiological agents while preserving the beneficial microbial diversity of the oral cavity. Targeting glucosyltransferases (GTFs), which play crucial roles in dental biofilm formation, offers a precise strategy to inhibit extracellular polysaccharide synthesis without compromising oral microbiota. This review article delves into the intricate mechanisms underlying dental caries, with a specific focus on the role of GTFs, enzymes produced by S. mutans. It further provides an overview of current research on GTF inhibitors, exploring their mechanisms of action, efficacy, and potential applications in clinical practice. Furthermore, it discusses the challenges and opportunities in the development of novel GTF inhibitors, emphasizing the need for innovative approaches to combat biofilm-mediated oral diseases effectively. [ABSTRACT FROM AUTHOR] more...
- Published
- 2024
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33. Immobilized glucosyltransferase and sucrose synthase on Fe3O4@Uio-66 in cascade catalysis for the one-pot conversion of rebaudioside D from rebaudioside A.
- Author
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Liu, Xiaojie, Hu, Yifan, Wei, Bin, Liu, Fang, Xu, Haichang, Liu, Changxia, Li, Ye, and Liang, Hao
- Subjects
- *
IRON oxides , *IMMOBILIZED enzymes , *ENZYME stability , *SUCROSE , *ENCAPSULATION (Catalysis) , *TRANSMISSION electron microscopy - Abstract
Rebaudioside D (RD) is a natural low-calorie, high-sweet sweetener and an excellent sugar substitute. In this study, RD was produced by coupling glucosyltransferase (EUGT11) and sucrose synthase (SUS) with a novel strategy of immobilization to enhance the recyclability of enzymes and decrease the production costs of RD. Recombinant EUGT11 from Oryza sativa and SUS from Arabidopsis thaliana were expressed in Escherichia coli and immobilized onto Fe 3 O 4 @Uio-66 nanocomposite. The results of Fourier transform Infrared spectra (FT-IR), X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), magnetic susceptibility (VSM), thermogravimetric (TG) analysis and Brunauer–Emmett–Teller (BET) indicated that the magnetic nanocomposite Fe 3 O 4 @Uio-66 was successfully fabricated and the two enzymes were separately immobilized on Fe 3 O 4 @Uio-66. The reusability, storage, pH and temperature stabilities of the immobilized enzymes were investigated and compared to that of free enzymes. It was more stable towards temperature compared with free enzyme. The kinetic properties of immobilized EUGT11 showed a lower V m and a higher K m compared to free EUGT11, and immobilized SUS showed a lower V m and K m compared to free SUS. The immobilized SUS had around 56% residual activity upon storage a period of 10 days at 4 °C. After 8 times, the catalytic activity of immobilized double enzyme still retained around 80%, which showed a desirable stability and reduced the overall cost of enzymes, indicating that the immobilized enzymes had a good industrial application prospect in RD production. [Display omitted] • Double-enzyme biocatalyst of glucosyltransferase and sucrose synthase was prepared. • Glucosyltransferase and sucrose synthase were first immobilized on Fe 3 O 4 @Uio-66. • The thermostability of enzyme was improved by immobilization. • The activity of immobilized double enzyme was maintained at 80% after 8 times of recycling. [ABSTRACT FROM AUTHOR] more...
- Published
- 2022
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34. Whole Genome Sequence Analysis of a Novel Apilactobacillus Species from Giant Honeybee (Apis dorsata) Gut Reveals Occurrence of Genetic Elements Coding Prebiotic and Probiotic Traits.
- Author
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Ahmad, Waqar, Khaliq, Shazia, Akhtar, Nasrin, El Arab, Jamilah, Akhtar, Kalsoom, Prakash, Satya, Anwar, Munir A., and Munawar, Nayla
- Subjects
PROBIOTICS ,WHOLE genome sequencing ,METHIONINE sulfoxide reductase ,GENETIC code ,SEQUENCE analysis ,HONEYBEES ,GENOMES - Abstract
Apilactobacillus spp. are classified as obligate fructophilic lactic acid bacteria (FLAB) that inhabit fructose-rich niches such as honeybee gut. Lactic acid bacteria are an important component of the gut microbiome and play a crucial role in maintaining gut health. In this study, a new FLAB strain HBW1, capable of producing glucan-type exopolysaccharide, was isolated from giant honeybee (Apis dorsata) gut and subjected to whole genome sequencing (WHS) to determine its health-beneficial traits. The genome size of the isolate was 1.49 Mb with a GC content of 37.2%. For species level identity, 16S rDNA sequence similarity, genome to genome distance calculator (dDDH), and average nucleotide identity (ANI) values were calculated. Phylogenetic analysis showed that the isolate HBW1 belongs to the Apilactobacillus genus. The dDDH and ANI values in comparison with closely clustered Apilactobacillus kunkeei species were 52% and 93.10%, respectively. Based on these values, we concluded that HBW1 is a novel species of Apilactobacillus, and we propose the name Apilactobacillus waqarii HBW1 for it. Further, WHS data mining of HBW1 revealed that it harbors two glucosyltransferase genes for prebiotic glucan-type exopolysaccharide synthesis. Moreover, chaperon (clp) and methionine sulfoxide reductase (msrA, msrB, and msrC) genes as well as nutritional marker genes for folic acid (folD) and riboflavin biosynthesis (rib operon), important for conferring probiotic properties, were also detected. Occurrence of these genetic traits make HBW1 an excellent candidate for application to improve gut function. [ABSTRACT FROM AUTHOR] more...
- Published
- 2022
- Full Text
- View/download PDF
35. Raffinose Inhibits Streptococcus mutans Biofilm Formation by Targeting Glucosyltransferase
- Author
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So-Young Ham, Han-Shin Kim, Eunji Cha, Taehyeung Lim, Youngjoo Byun, and Hee-Deung Park
- Subjects
raffinose ,Streptococcus mutans ,biofilm ,glucosyltransferase ,Microbiology ,QR1-502 - Abstract
ABSTRACT Streptococcus mutans is a representative biofilm-forming bacterium that causes dental caries through glucosyltransferase (GTF) activity. Glucans are synthesized from sucrose by GTFs and provide binding sites for S. mutans to adhere tightly to the tooth enamel. Therefore, if a novel compound that interferes with GTF function is developed, biofilm formation control in S. mutans would be possible. We discovered that raffinose, an oligosaccharide from natural products, strongly inhibited biofilm formation, GTF-related gene expression, and glucan production. Furthermore, biofilm inhibition on saliva-coated hydroxyapatite discs through the reduction of bacterial adhesion indicated the applicability of raffinose in oral health. These effects of raffinose appear to be due to its ability to modulate GTF activity in S. mutans. Hence, raffinose may be considered an antibiofilm agent for use as a substance for oral supplies and dental materials to prevent dental caries. IMPORTANCE Dental caries is the most prevalent infectious disease and is expensive to manage. Dental biofilms can be eliminated via mechanical treatment or inhibited using antibiotics. However, bacteria that are not entirely removed or are resistant to antibiotics can still form biofilms. In this study, we found that raffinose inhibited biofilm formation by S. mutans, a causative agent of dental caries, possibly through binding to GtfC. Our findings support the notion that biofilm inhibition by raffinose can be exerted by interference with GTF function, compensating for the shortcomings of existing commercialized antibiofilm methods. Furthermore, raffinose is an ingredient derived from natural products and can be safely utilized in humans; it has no smell and tastes sweet. Therefore, raffinose, which can control S. mutans biofilm formation, has been suggested as a substance for oral supplies and dental materials to prevent dental caries. more...
- Published
- 2022
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- View/download PDF
36. Eugenol functions as a signal mediating cold and drought tolerance via UGT71A59‐mediated glucosylation in tea plants.
- Author
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Zhao, Mingyue, Jin, Jieyang, Wang, Jingming, Gao, Ting, Luo, Yu, Jing, Tingting, Hu, Yutong, Pan, Yuting, Lu, Mengqian, Schwab, Wilfried, and Song, Chuankui
- Subjects
- *
DROUGHT tolerance , *URIDINE diphosphate , *REACTIVE oxygen species , *ABSCISIC acid , *TEA , *COLD (Temperature) - Abstract
SUMMARY Cold and drought stress are the most critical stresses encountered by crops and occur simultaneously under field conditions. However, it is unclear whether volatiles contribute to both cold and drought tolerance, and if so, by what mechanisms they act. Here, we show that airborne eugenol can be taken up by the tea (Camellia sinensis) plant and metabolized into glycosides, thus enhancing cold and drought tolerance of tea plants. A uridine diphosphate (UDP)‐glucosyltransferase, UGT71A59, was discovered, whose expression is strongly induced by multiple abiotic stresses. UGT71A59 specifically catalyzes glucosylation of eugenol glucoside in vitro and in vivo. Suppression of UGT71A59 expression in tea reduced the accumulation of eugenol glucoside, lowered reactive oxygen species (ROS) scavenging capacity, and ultimately impaired cold and drought stress tolerance. Exposure to airborne eugenol triggered a marked increase in UGT71A59 expression, eugenol glucoside accumulation, and cold tolerance by modulating ROS accumulation and CBF1 expression. It also promoted drought tolerance by altering abscisic acid homeostasis and stomatal closure. CBF1 and CBF3 play positive roles in eugenol‐induced cold tolerance and CBF2 may be a negative regulator of eugenol‐induced cold tolerance in tea plants. These results provide evidence that eugenol functions as a signal in cold and drought tolerance regulation and shed new light on the biological functions of volatiles in the response to multiple abiotic stresses in plants. Significance Statement: The volatile eugenol functions as a signal and plays a positive role in cold stress through regulating reactive oxygen species accumulation and CBF1 expression and in drought stress through altering abscisic acid homeostasis and stomatal closure via UDP‐glucosyltransferase UGT71A59‐mediated eugenol glucosylation in tea (Camellia sinensis) plants. [ABSTRACT FROM AUTHOR] more...
- Published
- 2022
- Full Text
- View/download PDF
37. Identification and characterization of unique 5-hydroxyisoflavonoid biosynthetic key enzyme genes in Lupinus albus.
- Author
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Liu, Jinyue and Jiang, Wenbo
- Subjects
- *
LUPINUS albus , *NICOTIANA benthamiana , *ISOFLAVONES , *ENZYMES , *ISOFLAVONOIDS , *BIOACTIVE compounds - Abstract
Key message: 5-Hydroxyisoflavonoids, no 5-deoxyisoflavonoids, in Lupinus species, are due to lack of CHRs and Type II CHIs, and the key enzymes of isoflavonoid biosynthetic pathway in white lupin were identified. White lupin (Lupinus albus) is used as food ingredients owing to rich protein, low starch, and rich bioactive compounds such as isoflavonoids. The isoflavonoids biosynthetic pathway in white lupin still remains unclear. In this study, only 5-hydroxyisoflavonoids, but no 5-deoxyisoflavonoids, were detected in white lupin and other Lupinus species. No 5-deoxyisoflavonoids in Lupinus species are due to lack of CHRs and Type II CHIs. We further found that the CHI gene cluster containing both Type I and Type II CHIs possibly arose after the divergence of Lupinus with other legume clade. LaCHI1 and LaCHI2 identified from white lupin metabolized naringenin chalcone to naringenin in yeast and tobacco (Nicotiana benthamiana), and were bona fide Type I CHIs. We further identified two isoflavone synthases (LaIFS1 and LaIFS2), catalyzing flavanone naringenin into isoflavone genistein and also catalyzing liquiritigenin into daidzein in yeast and tobacco. In addition, LaG6DT1 and LaG6DT2 prenylated genistein at the C-6 position into wighteone. Two glucosyltransferases LaUGT1 and LaUGT2 metabolized genistein and wighteone into its 7-O-glucosides. Taken together, our study not only revealed that exclusive 5-hydroxyisoflavonoids do exist in Lupinus species, but also identified key enzymes in the isoflavonoid biosynthetic pathway in white lupin. [ABSTRACT FROM AUTHOR] more...
- Published
- 2022
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38. Molecular cloning and functional characterization of an isoflavone glucosyltransferase from Pueraria thomsonii.
- Author
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DUAN, Hai-Yan, WANG, Jian, ZHA, Liang-Ping, PENG, Hua-Sheng, ZHAO, Yu-Ping, YUAN, Yuan, and HUANG, Lu-Qi
- Published
- 2022
- Full Text
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39. Cold Stress-induced Glucosyltransferase CsUGT78A15 is Involved in the Formation of Eugenol Glucoside in Camellia sinensis
- Author
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Mingyue Zhao, Binbin Cai, Jieyang Jin, Na Zhang, Tingting Jing, Jingming Wang, Yuting Pan, Zixiang Zhou, Yifan Zhao, Yingying Feng, Feng Yu, Mengting Zhang, Yating Li, Zhonghua Liu, and Chuankui Song
- Subjects
Camellia sinensis ,Eugenol ,Glucosyltransferase ,Cold stress ,Plant culture ,SB1-1110 - Abstract
Eugenol is a natural phenolic compound known for its health-promoting properties and its ability to add a floral scent to tea plants. Plant eugenol glycosides have been identified and shown to make important contributions to fruit floral quality. However, the details of their biosynthesis and metabolism in tea plants are still unknown. Here, eugenol glucoside was unambiguously identified as a native metabolite in the tea plant, and its biosynthesis was shown to be induced by low temperature treatment. Through the analysis of UGTs induced by low temperature, the glycosyltransferase CsUGT78A15 was identified in tea, and its encoded protein was shown to catalyze the glucosylation of eugenol. Vmax/Km ratios showed that eugenol was the most suitable substrate for CsUGT78A15. Sugar donor preference analysis showed that CsUGT78A15 had a higher selectivity for glucose, followed by galactose and glucuronic acid. The expression of CsUGT78A15 was correlated with the accumulation of eugenol glucoside in different tissues and genotypes of tea. Down-regulation of CsUGT78A15 led to a decreased eugenol glucoside content under cold stress, indicating that CsUGT78A15 plays an important role in the biosynthesis of eugenol glucoside under cold stress. The identification of eugenol glucoside in the tea plant and the discovery of a cold stress-induced eugenol glucosyltransferase in tea provide the foundation for the improvement of tea flavor under cold stress and the biotechnological production of eugenol glucoside. more...
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- 2020
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40. Synthesis of Polysaccharides III: Sucrase as Catalyst
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Kimura, Satoshi, Iwata, Tadahisa, He, Liang-Nian, Series Editor, Rogers, Robin D., Series Editor, Su, Dangsheng, Series Editor, Tundo, Pietro, Series Editor, Zhang, Z. Conrad, Series Editor, Kobayashi, Shiro, editor, Uyama, Hiroshi, editor, and Kadokawa, Jun-ichi, editor more...
- Published
- 2019
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41. A highly versatile fungal glucosyltransferase for specific production of quercetin-7-O-β-d-glucoside and quercetin-3-O-β-d-glucoside in different hosts.
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Ren, Jie, Tang, Wenzhu, Barton, Caleb Don, Price, Owen M., Mortensen, Mark Wayne, Phillips, Alexandra, Wald, Banner, Hulme, Simon Elgin, Stanley, Logan Powell, Hevel, Joan, and Zhan, Jixun
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BEAUVERIA bassiana , *PICHIA pastoris , *PSEUDOMONAS putida , *QUERCETIN , *NATURAL products , *RESVERATROL - Abstract
Glycosylation is an effective way to improve the water solubility of natural products. In this work, a novel glycosyltransferase gene (BbGT) was discovered from Beauveria bassiana ATCC 7159 and heterologously expressed in Escherichia coli. The purified enzyme was functionally characterized through in vitro enzymatic reactions as a UDP-glucosyltransferase, converting quercetin to five monoglucosylated and one diglucosylated products. The optimal pH and temperature for BbGT are 35 ℃ and 8.0, respectively. The activity of BbGT was stimulated by Ca2+, Mg2+, and Mn2+, but inhibited by Zn2+. BbGT enzyme is flexible and can glycosylate a variety of substrates such as curcumin, resveratrol, and zearalenone. The enzyme was also expressed in other microbial hosts including Saccharomyces cerevisiae, Pseudomonas putida, and Pichia pastoris. Interestingly, the major glycosylation product of quercetin in E. coli, P. putida, and P. pastoris was quercetin-7-O-β-d-glucoside, while the enzyme dominantly produced quercetin-3-O-β-d-glucoside in S. cerevisiae. The BbGT-harboring E. coli and S. cerevisiae strains were used as whole-cell biocatalysts to specifically produce the two valuable quercetin glucosides, respectively. The titer of quercetin-7-O-β-d-glucosides was 0.34 ± 0.02 mM from 0.83 mM quercetin in 24 h by BbGT-harboring E. coli. The yield of quercetin-3-O-β-d-glucoside was 0.22 ± 0.02 mM from 0.41 mM quercetin in 12 h by BbGT-harboring S. cerevisiae. This work thus provides an efficient way to produce two valuable quercetin glucosides through the expression of a versatile glucosyltransferase in different hosts. Key points: • A highly versatile glucosyltransferase was identified from B. bassiana ATCC 7159. • BbGT converts quercetin to five mono- and one di-glucosylated derivatives in vitro. • Different quercetin glucosides were produced by BbGT in E. coli and S. cerevisiae. [ABSTRACT FROM AUTHOR] more...
- Published
- 2022
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42. Identification and characterization of xanthone biosynthetic genes contributing to the vivid red coloration of red‐flowered gentian.
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Sasaki, Nobuhiro, Nemoto, Keiichirou, Nishizaki, Yuzo, Sugimoto, Naoki, Tasaki, Keisuke, Watanabe, Aiko, Goto, Fumina, Higuchi, Atsumi, Morgan, Ed, Hikage, Takashi, and Nishihara, Masahiro
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XANTHONE , *ANTHOCYANINS , *NUCLEAR magnetic resonance spectroscopy , *PROTEIN expression , *HIGH performance liquid chromatography , *WHEAT germ , *RECOMBINANT proteins - Abstract
Summary: Cultivated Japanese gentians traditionally produce vivid blue flowers because of the accumulation of delphinidin‐based polyacylated anthocyanins. However, recent breeding programs developed several red‐flowered cultivars, but the underlying mechanism for this red coloration was unknown. Thus, we characterized the pigments responsible for the red coloration in these cultivars. A high‐performance liquid chromatography with photodiode array analysis revealed the presence of phenolic compounds, including flavones and xanthones, as well as the accumulation of colored cyanidin‐based anthocyanins. The chemical structures of two xanthone compounds contributing to the coloration of red‐flowered gentian petals were determined by mass spectrometry and nuclear magnetic resonance spectroscopy. The compounds were identified as norathyriol 6‐O‐glucoside (i.e., tripteroside designated as Xt1) and a previously unreported norathyriol‐6‐O‐(6′‐O‐malonyl)‐glucoside (designated Xt2). The copigmentation effects of these compounds on cyanidin 3‐O‐glucoside were detected in vitro. Additionally, an RNA sequencing analysis was performed to identify the cDNAs encoding the enzymes involved in the biosynthesis of these xanthones. Recombinant proteins encoded by the candidate genes were produced in a wheat germ cell‐free protein expression system and assayed. We determined that a UDP‐glucose‐dependent glucosyltransferase (StrGT9) catalyzes the transfer of a glucose moiety to norathyriol, a xanthone aglycone, to produce Xt1, which is converted to Xt2 by a malonyltransferase (StrAT2). An analysis of the progeny lines suggested that the accumulation of Xt2 contributes to the vivid red coloration of gentian flowers. Our data indicate that StrGT9 and StrAT2 help mediate xanthone biosynthesis and contribute to the coloration of red‐flowered gentians via copigmentation effects. [ABSTRACT FROM AUTHOR] more...
- Published
- 2021
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43. EpsR Negatively Regulates Streptococcus mutans Exopolysaccharide Synthesis.
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Chen, J., Zhang, A., Xiang, Z., Lu, M., Huang, P., Gong, T., Pan, Y., Lin, Y., Zhou, X., and Li, Y.
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STREPTOCOCCUS mutans ,MICROBIAL exopolysaccharides ,DENTAL caries ,GLYCOSYLTRANSFERASES ,MICROBIOLOGICAL synthesis ,BIOFILMS - Abstract
Streptococcus mutans is considered the primary etiological agent of human dental caries. Glucosyltransferases (Gtfs) from S. mutans play important roles in the formation of biofilm matrix and the development of cariogenic oral biofilm. Therefore, Gtfs are considered an important target to prevent the development of dental caries. However, the role of transcription factors in regulating gtf expression is not yet clear. Here, we identify a MarR (multiple antibiotic resistance regulator) family transcription factor named EpsR (exopolysaccharide synthesis regulator), which negatively regulates gtfB expression and exopolysaccharide (EPS) production in S. mutans. The epsR in-frame deletion strain grew slowly, aggregated more easily in the presence of dextran, and displayed different colony morphology and biofilm structure. Notably, epsR deletion resulted in altered 3-dimensional biofilm architecture, increased water-insoluble EPS production, and upregulated GtfB protein content and activity. In addition, global gene expression profiling revealed differences in the expression levels of 69 genes in which gtfB was markedly upregulated. The conserved DNA motif for EpsR binding was determined by electrophoretic mobility shift assay and DNase I footprinting assays. Moreover, analysis of β-galactosidase activity suggested that EpsR acted as a repressor and inhibited gtfB expression. Taken together, our findings indicate that EpsR is an important transcription factor that regulates gtfB expression and EPS production in S. mutans. These results add new aspects to the complexity of regulating the expression of genes involved in the cariogenicity of S. mutans, which might lead to novel strategies to prevent the formation of cariogenic biofilm that may favor diseases. [ABSTRACT FROM AUTHOR] more...
- Published
- 2021
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44. Design and study of anticaries effect of different medicinal plants against S.mutans glucosyltransferase
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Kiranmai Mandava, Uma Rajeswari Batchu, Shravya Kakulavaram, Shulamithi Repally, Ishwarya Chennuri, Srinivas Bedarakota, and Namratha Sunkara
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S.mutans ,Glucosyltransferase ,Azadirachta indica ,Terminalia chebula ,Polyherbal mouth wash ,Anticaries agent ,Other systems of medicine ,RZ201-999 - Abstract
Abstract Background The present study was aimed to evaluate the molecular level anticaries effect of different medicinal plants against Streptococcus mutans (S.mutans) glucosyltransferases (gtf). Methods A total of six natural sources named as Terminalia chebula (T.chebula), Psidium guajava (P.guajava), Azadirachta indica (A.indica) and Pongamia pinnata (P.pinnata); two essential oils, clove (Syzygium aromaticum) and peppermint oil (Mentha piperita) were selected as test samples. Hydroalcoholic plant extracts and essential oils were examined for their inhibitory potential on gtf isolated from S.mutans. Polyherbal mouth wash was prepared and its effect on gtf activity was compared with commercial chlorhexidine mouth wash (5%w/v). Enzyme kinetic study was carried out in order to explore the molecular mechanism of enzyme action. Results Out of six natural sources tested, A.indica has shown maximum inhibitory effect of 91.647% on gtf and T.chebula has shown IC50 of 1.091 mg/ml which is significant when compared to standard chlorhexidine. From the final result of kinetic analysis it was found that T.chebula, P.guajava and P.pinnata have show uncompetitive inhibition where as A.indica has shown non-competitive inhibition. Surprisingly, both essential oils have shown allosteric inhibition (sigmoidal response). The polyherbal moutwash has shown significant inhibitory potential on gtf (95.936%) when compared to commercial chlorhexidine mouthwash (p more...
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- 2019
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45. Structural Aspects of ER Glycoprotein Quality-Control System Mediated by Glucose Tagging
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Satoh, Tadashi, Kato, Koichi, COHEN, IRUN R., Series Editor, LAJTHA, ABEL, Series Editor, LAMBRIS, JOHN D., Series Editor, PAOLETTI, RODOLFO, Series Editor, Rezaei, Nima, Series Editor, Yamaguchi, Yoshiki, editor, and Kato, Koichi, editor more...
- Published
- 2018
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46. High-level de novo biosynthesis of glycosylated zeaxanthin and astaxanthin in Escherichia coli.
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Chen, Xixian, Lim, Xiaohui, Bouin, Aurélie, Lautier, Thomas, and Zhang, Congqiang
- Subjects
ZEAXANTHIN ,ASTAXANTHIN ,BIOSYNTHESIS ,ESCHERICHIA coli ,CAROTENOIDS ,GLUCOSIDES - Abstract
Because of wide applications in food, feed, pharmaceutical and cosmetic industries, the carotenoid market is growing rapidly. Most carotenoids are hydrophobic, which limits their bioavailability. Glycosylation is a natural route that substantially increases the water solubility, as well as the bioavailability, photostability and biological activities of carotenoids. Here, we report metabolic engineering efforts (e.g., promoter and RBS engineering, optimization of carbon sources and supplementation of bottleneck genes) to produce glycosylated carotenoids in Escherichia coli. By fine-tuning the carotenoid-biosynthetic genes (crtX, crtZ and crtY), our strain produced up to 47.2 mg/L (~ 11,670 ppm) of zeaxanthin glucosides, ~ 78% of the total carotenoids produced. In another construct with mevalonate, astaxanthin pathway and crtX genes, the strain produced a mixture of carotenoid glucosides including astaxanthin and adonixanthin glucosides with a total yield of 8.1 mg/L (1774 ppm). Our work demonstrated a proof-of-concept study for the microbial biosynthesis of glycosylated carotenoids. [ABSTRACT FROM AUTHOR] more...
- Published
- 2021
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47. Genetic mapping identifies a rice naringenin O‐glucosyltransferase that influences insect resistance.
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Yang, Zhongyan, Li, Nana, Kitano, Takashige, Li, Ping, Spindel, Jennifer E., Wang, Lishuo, Bai, Genxiang, Xiao, Yiying, McCouch, Susan R., Ishihara, Atsushi, Zhang, Jili, Yang, Xin, Chen, Zepeng, Wei, Jianyu, Ge, Honghua, Jander, Georg, and Yan, Jian more...
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GENE mapping , *COTTON , *PLANT resistance to insects , *NARINGENIN , *SPODOPTERA littoralis , *NILAPARVATA lugens - Abstract
SUMMARY: Naringenin, the biochemical precursor for predominant flavonoids in grasses, provides protection against UV damage, pathogen infection and insect feeding. To identify previously unknown loci influencing naringenin accumulation in rice (Oryza sativa), recombinant inbred lines derived from the Nipponbare and IR64 cultivars were used to map a quantitative trait locus (QTL) for naringenin abundance to a region of 50 genes on rice chromosome 7. Examination of candidate genes in the QTL confidence interval identified four predicted uridine diphosphate‐dependent glucosyltransferases (Os07g31960, Os07g32010, Os07g32020 and Os07g32060). In vitro assays demonstrated that one of these genes, Os07g32020 (UGT707A3), encodes a glucosyltransferase that converts naringenin and uridine diphosphate‐glucose to naringenin‐7‐O‐β‐d‐glucoside. The function of Os07g32020 was verified with CRISPR/Cas9 mutant lines, which accumulated more naringenin and less naringenin‐7‐O‐β‐d‐glucoside and apigenin‐7‐O‐β‐d‐glucoside than wild‐type Nipponbare. Expression of Os12g13800, which encodes a naringenin 7‐O‐methyltransferase that produces sakuranetin, was elevated in the mutant lines after treatment with methyl jasmonate and insect pests, Spodoptera litura (cotton leafworm), Oxya hyla intricata (rice grasshopper) and Nilaparvata lugens (brown planthopper), leading to a higher accumulation of sakuranetin. Feeding damage from O. hyla intricata and N. lugens was reduced on the Os07g32020 mutant lines relative to Nipponbare. Modification of the Os07g32020 gene could be used to increase the production of naringenin and sakuranetin rice flavonoids in a more targeted manner. These findings may open up new opportunities for selective breeding of this important rice metabolic trait. Significance Statement: Quantitative trait locus mapping and targeted mutagenesis identified a glucosyltransferase, Os07g32020, that influences the relative abundance of naringenin, sakuranetin and other flavonoids in rice. Enhanced flavonoid production was associated with plant resistance to rice insect pests Spodoptera litura, Oxya hyla intricata and Nilaparvata lugens. These findings may open up new opportunities for selective breeding of this important rice metabolic trait. [ABSTRACT FROM AUTHOR] more...
- Published
- 2021
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48. Dissecting the Role of VicK Phosphatase in Aggregation and Biofilm Formation of Streptococcus mutans.
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Wang, S., Long, L., Yang, X., Qiu, Y., Tao, T., Peng, X., Li, Y., Han, A., Senadheera, D.B., Downey, J.S., Goodman, S.D., Zhou, X., and Cvitkovitch, D.G.
- Subjects
DENTAL caries ,HISTIDINE kinases ,GLYCOSYLTRANSFERASES ,POLYSACCHARIDES ,GENETIC regulation ,AGGLUTININS ,STREPTOCOCCUS mutans - Abstract
VicRK (WalRK or YycFG) is a conserved 2-component regulatory system (TCS) that regulates cell division, cell wall biosynthesis, and homeostasis in low-GC Gram-positive bacteria. VicRK is also associated with biofilm formation of Streptococcus mutans on the tooth surface as it directly regulates the extracellular polysaccharide (EPS) synthesis. Of the 2 components, VicK possesses both autokinase and phosphatase activities, which regulate the phosphorylation and dephosphorylation of the regulator VicR in response to environmental cues. However, the dual mechanism of VicK as the autokinase/phosphatase in regulating S. mutans ' responses is not well elucidated. Previously, it has been shown that the phosphatase activity depends on the PAS domain and residues in the DHp domain of VicK in S. mutans. Specifically, mutating proline at 222 in the PAS domain inhibits VicK phosphatase activity. We generated a VicK
P222A mutant to determine the level of VicR-P in the cytoplasm by Phos-tag sodium dodecyl sulfate polyacrylamide gel electrophoresis. We show that in VicKP222A phosphatase, attenuation increased phosphorylated VicR (VicR-P) that downregulated glucosyltransferases, gtfBC, thereby reducing the synthesis of water-insoluble polysaccharides (WIS-EPS) in the biofilm. In addition, VicKP222A presented as long-rod cells, reduced growth, and displayed asymmetrical division. A major adhesin of S. mutans, SpaP was downregulated in VicKP222A , making it unable to agglutinate in saliva. In summary, we have confirmed that VicK phosphatase activity is critical to maintain optimal phosphorylation status of VicR in S. mutans, which is important for cell growth, cell division, EPS synthesis, and bacterial agglutination in saliva. Hence, VicK phosphatase activity may represent a promising target to modulate S. mutans ' pathogenicity. [ABSTRACT FROM AUTHOR] more...- Published
- 2021
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49. Isolation and Functional Analysis of Genes Involved in Polyacylated Anthocyanin Biosynthesis in Blue Senecio cruentus
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Chenfei Lu, Yajun Li, Yumeng Cui, Jiangshan Ren, Fangting Qi, Jiaping Qu, He Huang, and Silan Dai
- Subjects
blue flower ,polyacylated anthocyanins ,Senecio cruentus ,acyltransferase ,glucosyltransferase ,Plant culture ,SB1-1110 - Abstract
Polyacylated anthocyanins with multiple glycosyl and aromatic acyl groups tend to make flowers display bright and stable blue colours. However, there are few studies on the isolation and functional characterization of genes involved in the polyacylated anthocyanin biosynthesis mechanism, which limits the molecular breeding of truly blue flowers. Senecio cruentus is an important potted ornamental plant, and its blue flowers contain 3′,7-polyacylated delphinidin-type anthocyanins that are not reported in any other plants, suggesting that it harbours abundant gene resources for the molecular breeding of blue flowers. In this study, using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis of blue, carmine and white colours of cineraria cultivars “Venezia” (named VeB, VeC, and VeW, respectively), we found that 3′,7-polyacylated anthocyanin, cinerarin, was the main pigment component that determined the blue colour of ray florets of cineraria. Based on the transcriptome sequencing and differential gene expression (DEG) analysis combined with RT- and qRT-PCR, we found two genes encoding uridine diphosphate glycosyltransferase, named ScUGT1 and ScUGT4; two genes encoding acyl-glucoside-dependent glucosyltransferases which belong to glycoside hydrolase family 1 (GH1), named ScAGGT11 and ScAGGT12; one gene encoding serine carboxypeptidase-like acyltransferase ScSCPL2; and two MYB transcriptional factor genes ScMYB2 and ScMYB4, that were specifically highly expressed in the ray florets of VeB, which indicated that these genes may be involved in cinerarin biosynthesis. The function of ScSCPL2 was analysed by virus-induced gene silencing (VIGS) in cineraria leaves combined with HPLC-MS/MS. ScSCPL2 mainly participated in the 3′ and 7-position acylation of cinerarin. These results will provide new insight into the molecular basis of the polyacylated anthocyanin biosynthesis mechanism in higher plants and are of great significance for blue flower molecular breeding of ornamental plants. more...
- Published
- 2021
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50. Two UGT84A Family Glycosyltransferases Regulate Phenol, Flavonoid, and Tannin Metabolism in Juglans regia (English Walnut)
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Houston J. Saxe, Takanori Horibe, Bipin Balan, Timothy S. Butterfield, Noah G. Feinberg, Christopher M. Zabaneh, Aaron E. Jacobson, and Abhaya M. Dandekar
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glucosyltransferase ,hydrolysable tannin ,UGT84A ,gallic acid ,walnut ,phenol metabolism ,Plant culture ,SB1-1110 - Abstract
We showed previously that gallic acid is produced in walnut from 3-dehydroshikimate by a shikimate dehydrogenase (JrSkDH). This study focuses on the next step in the hydrolysable tannin pathway, the formation of 1-O-galloyl-β-D-glucose from the phenolic gallic acid and UDP glucose by a glycosyltransferase. JrGGT1 (UGT84A73) and JrGGT2 (UGT84A74) are predicted to be two such glycosyltransferases, which we expressed in tobacco plants. GC-MS analysis of the transgenic tobacco revealed moderate, yet significant alterations in plant secondary metabolism, such as depleted phenolic acids, including gallic acid. We postulate that these effects are due to JrGGT1 and JrGGT2 activity, as JrGGT orthologs glycosylate these phenolic compounds in vitro. Moreover, JrGGT expression in tobacco caused upregulation of shikimic acid pathway metabolites and differing responses in phenylpropanoids, such as phenolic acids and flavonoids. In transcriptome analysis of walnut pellicle tissues, both JrGGTs showed substantial and significant expression correlations with the gallic acid-producing JrSkDHs and were highly coexpressed with the genetic circuits constituting the shikimic acid and phenylpropanoid biosynthetic pathways. Verification of JrGGT gene expression by transcriptome analysis of 20 walnut tissues revealed striking similarities with that of the pellicle data, with the greatest expression in roots, wood, buds, and leaves of Juglans regia cv. Chandler: tissues that typically accumulate hydrolysable tannins. Like the transgenic tobacco, pellicle metabolomic analyses revealed that many phenylpropanoids correlated negatively with JrGGT expression, while shikimic acid pathway metabolites correlated positively with JrGGT expression. This research supports the hypothesis that JrGGT1 and JrGGT2 play non-trivial roles in metabolism of phenolic acids, flavonoids, and ostensibly, tannins. more...
- Published
- 2021
- Full Text
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