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SUMMARY Previously, in Arabidopsis thaliana, we found atypical spherical starch granules in dpe2ss4 and dpe2phs1ss4. However, the mechanism of such abnormal morphogenesis is still obscure. By tracking starch granule length and thickness with leaf ageing, we reported that the starch granules in dpe2phs1ss4 gradually change to a spherical shape over time. In comparison, Col‐0 and the parental line ss4 did not exhibit macroscopic morphological alteration. In this study, firstly, we specify that the additional lack of DPE2 resulted in the gradual alteration of starch granule morphology over time. Similar gradual morphological alterations were also found in dpe2, mex1, and sex4 but not in the other starch degradation‐related mutants, such as sex1‐8, pwd, and bam3. The gradual alteration of starch morphology can be eliminated by omitting the dark phase, suggesting that the particular impaired starch degradation in dpe2‐ and mex1‐related mutants influences starch morphology. Secondly, we observed that spherical starch morphology generation was accompanied by prominent elevated short glucan chains of amylopectin and an increased amylose proportion. Thirdly, the interplay between soluble starch synthase 2 and branching enzymes was affected and resulted in the formation of spherical starch granules. The resulting spherical starch granules allow for elevated starch synthesis efficiency. Fourthly, the starch phosphate content at the granule surface correlated with the morphology alteration of the starch granules. Herewith, we propose a model that spherical starch granules, accumulated in mutants with a misbalance of the starch degradation pathway, are result of elevated starch synthesis to cope with overloaded carbohydrates. [ABSTRACT FROM AUTHOR]

In this study, growth and enzyme production of Bacillus megaterium RB-05 was achieved at varying parameters such as temperature, incubation time, pH, carbon source and nitrogen source. Naturally occurring sand with 97% quartz was utilized as the matrix for bacterial growth and CaCO3 precipitation. The estimation of CaCO3 precipitation was done through gravimetric analysis and CaCO3 deposition was detected through EDX and SEM analysis. Furthermore, sand stabilization was determined through Drained triaxial tests. The results obtained showed maximum bacterial growth and urease activity at pH 7, 350C temperature and 48 h incubation time. Bacteria showed maximum growth with glucose as carbon source however, significant urease activity was observed with both glucose and maltose. Optimum bacterial growth was observed with ammonium chloride as the nitrogen source however, maximum urease activity was observed with ammonium sulphate. CaCO3 precipitation of treated sand showed a linear relation between CaCO3 precipitation with urease production. The EDX and SEM analysis showed CaCO3 deposition between sand particles and on the surface. Furthermore, the stress response of sand showed enhanced sand strength at maximum CaCO3 precipitation. The obtained results show the link between urease production, CaCO3 precipitation, and sand stabilization by MICP. High urease activity influences CaCO3 precipitation in the soil followed by enhanced soil strength. However, the positive results were obtained in reactor environment, further studies are required to make the technique applicable. [ABSTRACT FROM AUTHOR]

Purpose: Iron deficiency anemia is common in people with inflammatory bowel disease (IBD), causing deterioration in quality of life, which can be reversed by treatment that increases iron stores and hemoglobin levels. The present post hoc analyses estimate health state utility values for patients with IBD after treatment with ferric derisomaltose or ferric carboxymaltose and evaluate the health domains driving the changes. Methods: SF-36v2 responses were recorded at baseline and day 14, 35, 49, and 70 from 97 patients enrolled in the randomized, double-blind, PHOSPHARE-IBD trial (ClinicalTrials.gov ID: NCT03466983), in which patients with IBD across five European countries were randomly allocated to either ferric derisomaltose or ferric carboxymaltose. Changes in SF-36v2 scale scores and SF-6Dv2 health utility values were analyzed by mixed models. Results: In both treatment arms, SF-6Dv2 utility values and all SF-36v2 scale scores, except Bodily Pain, improved significantly (p = < 0.0001). The improvement in SF-6Dv2 utility values showed no significant treatment group difference. The improvement in utility values was completely explained by improvement in Vitality scores. Vitality scores showed significantly larger improvement with ferric derisomaltose versus ferric carboxymaltose (p = 0.026). Patients with the smallest decrease in phosphate had significantly larger improvements in Vitality scores at each time point (p = < 0.05 for all comparisons) and overall (p = 0.0006). Conclusions: Utility values improved significantly with intravenous iron treatment. Improvement in utility values was primarily driven by Vitality scores, which showed significantly greater improvement in the ferric derisomaltose arm. Smaller decreases in phosphate were associated with significantly higher Vitality scores, suggesting that quality of life improvement is attenuated by hypophosphatemia. The utility values can inform future cost-utility analysis. [ABSTRACT FROM AUTHOR]

This study aimed to experimentally investigate the effect of sugar syrup additions on quality measurements of honey and to detect adulteration. For that purpose, two different pure blossom honey samples were adulterated by directly mixing 0%, 5%, 10%, 20%, 30%, 40%, and 50% of commercially available glucose–fructose corn syrup and maltose corn syrup. In this regard, key physico-chemical properties like moisture, pH, free acidity, proline, diastase number, color (L, a, b and Delta-E), electrical conductivity, HMF, sugar profile (glucose, fructose, sucrose and maltose), and C4 sugar analysis were tested. The results of the individual analysis of moisture, pH, free acidity, proline, diastase number, color, electrical conductivity, and HMF failed to detect sugar syrup adulteration. However, when principal component analysis (PCA) was utilized to analyze the data gathered from these tests, adulterations at all-syrup ratios (5–50%) were successfully detected. [ABSTRACT FROM AUTHOR]

Nutrient signaling pathways play a pivotal role in regulating the balance among metabolism, growth and stress response depending on the available food supply. They are key factors for the biotechnological success of the yeast Saccharomyces cerevisiae during food-producing fermentations. One such pathway is Retrograde Response, which controls the alpha-ketoglutarate supply required for the synthesis of amino acids like glutamate and lysine. Repressor MKS1 is linked with the TORC1 complex and negatively regulates this pathway. Deleting MKS1 from a variety of industrial strains causes glycerol to increase during winemaking, brewing and baking. This increase is accompanied by a reduction in ethanol production during grape juice fermentation in four commercial wine strains. Interestingly, this does not lead volatile acidity to increase because acetic acid levels actually lower. Aeration during winemaking usually increases acetic acid levels, but this effect reduces in the MKS1 mutant. Despite the improvement in the metabolites of oenological interest, it comes at a cost given that the mutant shows slower fermentation kinetics when grown in grape juice, malt and laboratory media and using glucose, sucrose and maltose as carbon sources. The deletion of RTG2 , an activator of Retrograde Response that acts as an antagonist of MKS1 , also results in a defect in wine fermentation speed. These findings suggest that the deregulation of this pathway causes a fitness defect. Therefore, manipulating repressor MKS1 is a promising approach to modulate yeast metabolism and to produce low-ethanol drinks. • MKS1 gene deletion increases glycerol and reduces ethanol during winemaking. • This increase in glycerol does not imply an overproduction of acetic acid. • Glycerol also increases in brewing and baking yeasts. [ABSTRACT FROM AUTHOR]

Dextransucrases play a crucial role in the production of dextran from economical sucrose; therefore, there is a pressing demand to explore novel dextransucrases with better performance. This study characterized a dextransucrase enzyme, LmDexA, which was identified from the Leuconostoc mesenteroides NN710. This bacterium was isolated from the soil of growing dragon fruit in Guangxi province, China. We successfully constructed six different N-terminal truncated variants through sequential analysis. Additionally, a truncated variant, ΔN190LmDexA, was constructed by removing the 190 amino acids fragment from the N-terminal. This truncated variant was then successfully expressed heterologously in Escherichia coli and purified. The purified ΔN190LmDexA demonstrated optimal hydrolysis activity at a pH of 5.6 and a temperature of 30 °C. Its maximum specific activity was measured to be 126.13 U/mg, with a Km of 13.7 mM. Results demonstrated a significant improvement in the heterologous expression level and total enzyme activity of ΔN190LmDexA. ΔN190LmDexA exhibited both hydrolytic and transsaccharolytic enzymatic activities. When sucrose was used as the substrate, it primarily produced high-molecular-weight dextran (>400 kDa). However, upon the addition of maltose as a receptor, it resulted in the production of a significant amount of oligosaccharides. Our results can provide valuable information for enhancing the characteristics of recombinant dextransucrase and potentially converting sucrose into high-value-added dextran and oligosaccharides. [ABSTRACT FROM AUTHOR]

Diabetes is a multifactorial metabolic disorder occurs due to uncontrolled persistent hyperglycaemia. The α-glucosidase enzyme plays an important role in management of diabetes. The α-glucosidase enzyme gets secreted by the brush border cells of small intestine which helps in converting maltose into glucose and thereby inhibiting the enzyme will help in lowering blood glucose level. In the present study, 100 compounds were selected having activity against α-glucosidase enzyme and they were used to build a machine learning based quantitative structure activity relationship model (QSAR). All the compounds were having thiazolidinedione (TZD) as the common nucleus. The molecules selected were divided into training and testing datasets of 80:20 ratio for various model development. The important molecular descriptors which will affect the target were chosen using recursive feature elimination (RFE) algorithm. The predictive models were created using machine learning regression techniques including Support Vector Regression (SVR), Random Forest Regression (RFR), Decision Tree Regression (DTR) and Gradient Boosting Regression (GBR). A comparison-based analysis was done between the various machine learning algorithms. The GBR and RFR gave the best R² value of 0.9992 and 0.9514 for the training dataset and 0.9414 and 0.8760 for the testing dataset respectively, followed by SVR and DTR. Thus, it concludes that the four-machine learning algorithm generates a highly predictive model for the unique compounds and a superior prediction capability for building a QSAR model for α-glucosidase enzyme inhibitors. [ABSTRACT FROM AUTHOR]

[Objective] The study aimed to identify the species of the pathogen of Kadsura coccinea (Lem.) A. C. Smith, screen out suitable fungicides, and provide theoretical basis for the identification and field prevention of the pathogen of K. coccinea. [Method] The pathogen of K. coccinea leaves collected from Yunfu, Guangdong province were isolated by tissue block separation method, which was determined based on Koch's postulates and was identified by morphology and molecular biology. The mycelial growth rate method was used for biological testing, as well as indoor toxicity of different fungicides to K. coccinea. [Result] By isolating and purifying the diseased leaves, a total of 6 strains were obtained. And representative strain Al was selected for morphological identification and ITS sequence analysis. The pathogen of K. coccinea was identified as Colletotrichum sp.. Through biological testing of strain Al, it was determined that the most suitable growth medium was PDA medium, the most suitable temperature for mycelial growth was 28 °C, the lethal temperature was 53 °C for 10 minutes, the optimal pH for mycelium growth was 6.0, the optimal lighting conditions were continuous darkness, the optimal carbon and nitrogen sources for this mycelium were maltose and yeast powder. Among the four tested fungicides of thiram, bromoxynil, carbendazim and azoxystrobin, carbendazim had the best inhibitory effect, with an EC50 value of 0. 14 µg/mL.. Followed by thiram and bromoxynil, with EC50 values of 55. 26 and 55.32 µg/ml., respectively, indicating that they had better inhibitory that effect. The EC50 value of azoxystrobin was 791.95 µg/mL, with a range of 100-1000 µg/mL., and the antibacterial effect was not significant. [Conclusion] The results show that Colletotrichum sp. is the pathogen of Kadsura coccinea collected from Yunfu, Guangdong province. Carbendazim, thiram and bromoxynil could be used as candidate field control agents for the pathogen of K. coccinea. [ABSTRACT FROM AUTHOR]

Lager yeasts are limited to a few strains worldwide, imposing restrictions on flavour and aroma diversity and hindering our understanding of the complex evolutionary mechanisms during yeast domestication. The recent finding of diverse S. eubayanus lineages from Patagonia offers potential for generating new lager yeasts with different flavour profiles. Here, we leverage the natural genetic diversity of S. eubayanus and expand the lager yeast repertoire by including three distinct Patagonian S. eubayanus lineages. We used experimental evolution and selection on desirable traits to enhance the fermentation profiles of novel S. cerevisiae x S. eubayanus hybrids. Our analyses reveal an intricate interplay of pre-existing diversity, selection on species-specific mitochondria, de-novo mutations, and gene copy variations in sugar metabolism genes, resulting in high ethanol production and unique aroma profiles. Hybrids with S. eubayanus mitochondria exhibited greater evolutionary potential and superior fitness post-evolution, analogous to commercial lager hybrids. Using genome-wide screens of the parental subgenomes, we identified genetic changes in IRA2, IMA1, and MALX genes that influence maltose metabolism, and increase glycolytic flux and sugar consumption in the evolved hybrids. Functional validation and transcriptome analyses confirmed increased maltose-related gene expression, influencing greater maltotriose consumption in evolved hybrids. This study demonstrates the potential for generating industrially viable lager yeast hybrids from wild Patagonian strains. Our hybridization, evolution, and mitochondrial selection approach produced hybrids with high fermentation capacity and expands lager beer brewing options. Author summary: Lager beer dominates the global market, accounting for over 90% of commercial beer varieties. The main player in lager fermentation is the yeast Saccharomyces pastorianus, an interspecific hybrid between S. cerevisiae and S. eubayanus. Despite its popularity, the range of flavours and aromas found in lager beers is restricted by the low genetic diversity of available lager strains. Here, we explored if lager yeast profiles can be diversified by leveraging natural isolates of S. eubayanus from Chilean Patagonia. We generated de novo hybrids between S. cerevisiae and three distinct S. eubayanus Patagonian lineages. Through experimental evolution and selection on fermentation traits, we improved the fermentation profiles of the hybrids. We found that mutations in IRA2, IMA1, and MALX genes enhanced their maltose and maltotriose metabolism, resulting in higher ethanol production and unique aroma profiles. Our results also confirm that S. eubayanus mitochondria confer a greater evolutionary potential than S. cerevisiae mitochondria. The current study encourages the use of wild yeast strains to develop new brewing applications to expand the repertoire of de novo lager yeasts. [ABSTRACT FROM AUTHOR]

Periplasmic binding proteins (PBPs) are bacterial proteins commonly used as scaffolds for substrate-detecting biosensors. In these biosensors, effector proteins (for example fluorescent proteins) are inserted into a PBP such that the effector protein's output changes upon PBP-substate binding. The insertion site is often determined by comparison of PBP apo/holo crystal structures, but random insertion libraries have shown that this can miss the best sites. Here, we present a PBP biosensor design method based on residue contact analysis from molecular dynamics. This computational method identifies the best previously known insertion sites in the maltose binding PBP, and suggests further previously unknown sites. We experimentally characterise fluorescent protein insertions at these new sites, finding they too give functional biosensors. Furthermore, our method is sufficiently flexible to both suggest insertion sites compatible with a variety of effector proteins, and be applied to binding proteins beyond PBPs. Author summary: "Biosensors" are microscopic tools that can detect specific molecules of interest and are made of biological building blocks, such as proteins. Upon coming into contact with their target molecule, such as a marker of a specific disease, biosensors change shape and their properties are altered. For example, upon contact with a disease marker, a biosensor could glow brightly to work as a diagnostic, or perhaps it could produce a chemical signal that can be detected by the immune system. Understanding how biosensors change shape in response to their target is key to developing more biosensors with complex responses. In this paper, we use computer simulations of a biosensor component to understand what parameters are important for biosensor design. We use these parameters to generate new biosensors that respond to their target molecule by producing light, but put forward a case that the lesson learned are generalisable enough to inform more complex sensor outputs as well. [ABSTRACT FROM AUTHOR]

α-Amylase plays a crucial role in the industrial degradation of starch. The genus Jeotgalibacillus of the underexplored marine bacteria family Caryophanaceae has not been investigated in terms of α-amylase production. Herein, we report the comprehensive analysis of an α-amylase (AmyJM) from Jeotgalibacillus malaysiensis D5T (= DSM28777T = KCTC33550T). Protein phylogenetic analysis indicated that AmyJM belongs to glycoside hydrolase family 13 subfamily 5 (GH13_5) and exhibits low sequence identity with known α-amylases, with its closest counterpart being the GH13_5 α-amylase from Bacillus sp. KSM-K38 (51.05% identity). Purified AmyJM (molecular mass of 70 kDa) is stable at a pH range of 5.5–9.0 and optimally active at pH 7.5. The optimum temperature for AmyJM is 40 °C, where the enzyme is reasonably stable at this temperature. Similar to other α-amylases, the presence of CaCl2 enhanced both the activity and stability of AmyJM. AmyJM exhibited activity toward raw and gelatinized forms of starches and related α-glucans, generating a mixture of reducing sugars, such as glucose, maltose, maltotriose, maltotetraose, and maltopentaose. In raw starch hydrolysis, AmyJM exhibited its highest efficiency (51.10% degradation) in hydrolyzing raw wheat starch after 3-h incubation at 40 °C. Under the same conditions, AmyJM also hydrolyzed tapioca, sago, potato, rice, and corn raw starches, yielding 16.01–30.05%. These findings highlight the potential of AmyJM as a biocatalyst for the saccharification of raw starches, particularly those derived from wheat. Key points: A novel GH13_5 α-amylase (AmyJM) from Jeotgalibacillus malaysiensis D5T was successfully expressed, purified, and biochemically characterized. AmyJM hydrolyzed raw starches from wheat, tapioca, sago, potato, rice, and corn at 40 °C, forming mixtures of reducing sugars. The properties of AmyJM suggest that the enzyme is potentially applicable in direct raw starch degradation processes. [ABSTRACT FROM AUTHOR]

Background: Variability in sugar content between raw and cooked sweetpotato storage roots impact nutritional and dietary importance with implications for consumer preference. High‐throughput phenotyping is required to breed varieties that satisfy consumer preferences. Results: Near‐infrared reflectance spectroscopy (NIRS) calibration curves were developed for analysing sugars in baked storage roots using 147 genotypes from a population segregating for sugar content and other traits. The NIRS prediction curves had high coefficients of determination in calibration (R2c) of 0.96 (glucose), 0.93 (fructose), 0.96 (sucrose), and 0.96 (maltose). The corresponding coefficients of determination for cross‐validation (R2cv) were 0.92 (glucose), 0.89 (fructose), 0.96 (sucrose) and 0.93 (maltose) and were similar to the R2c for all sugars measured. The ratios of the standard deviation of the reference set to the standard error of cross‐validation were greater than three for all sugars. These results confirm the applicability of the NIRS curves in efficiently determining sugar content in baked sweetpotato storage roots. External validation was performed on an additional 70 genotypes. Coefficients of determination (r2) were 0.88 (glucose), 0.88 (fructose), 0.86 (sucrose) and 0.49 (maltose). The results were comparable to those found for the calibration and cross‐validation in fructose, glucose, and sucrose, but were moderate for maltose due to the low variability of maltose content in the population. Conclusions: NIRS can be used for screening sugar content in baked sweetpotato storage roots in breeding programs and can be used to assist with the development of improved sweetpotato varieties that better meet consumer preferences. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Doubled haploid (DH) production technology has been a valuable tool for the development of homozygous lines through anther culture within a short period in comparison with conventional breeding methods. Doubled haploid production in pearl millet through anther culture helps to overcome inbreeding depression which occurs due to continuous selfing for homozygous line development. We developed an efficient method to develop DH plants through anther culture in pearl millet in large numbers, which is the first report in pearl millet. The method encompasses a selection of the right stage of mother plant, optimal culture media, and influence of temperature for treatment of anther for callus induction followed by plant regeneration and flow cytometric analysis for ploidy determination. Anther culture experiments were done in proprietary pearl millet genotype of Rasi seeds '1988F1' (R × R cross) for achieving better calli or embryo induction and green plant regeneration in pearl millet. A significant frequency of 35.4% calli-embryoid response in Doubled-haploid Pearl millet Media (DPM4) was achieved at incubation temperature of 32 °C for spikes pre-treatment and anther initiation. This response was further improved to 44.4% by separating anthers from spikes on fourth day of incubation. Alteration of basal salt concentration and inclusion of 12% maltose as carbon source in calli induction media resulted in a higher number of embryo-like structures (ELS) which eventually increased the regeneration up to 12.9% in Regeneration Media (RM3). These experiments resulted in a promising procedure to develop DH plants in pearl millet to minimize the timeline for pure line development and trait introgression to accelerate breeding program. [ABSTRACT FROM AUTHOR]

A method for quantitative determination of carbohydrate stabilizers (sorbitol, mannitol, glucose, sucrose, lactose, maltose, and trehalose) in biologics was developed. The analysis used HPLC with refractometric detection and a mobile phase of 75% MeCN in H2O. The developed method was specific and linear in the range 0.5 to 2 ng/mL. The coefficients of determination of the calibration curves were 0.99996 for sorbitol, 0.99998 for mannitol, 0.99971 for glucose, 0.99996 for sucrose, 0.99994 for lactose, 9.99966 for maltose monohydrate, and 0.99996 for trehalose dihydrate. The limits of quantitation (mg/mL) were 0.01 for sorbitol, 0.03 for mannitol, 0.05 for glucose, 0.025 for sucrose, 0.050 for lactose, 0.05 for maltose monohydrate, and 0.25 for trehalose dihydrate. The average accuracy covered the range (100 ± 5)%. [ABSTRACT FROM AUTHOR]

Objective: To study the transport activity of bacterial rhodopsins, proteoliposomes with the same orientation of the protein molecules should be obtained. Methods: The genes of fusion proteins containing Exiguobacterium sibiricum proteorhodopsin (ESR) and various N-terminal soluble domains have been constructed. Results and Discussion: Effective synthesis in Escherichia coli cells was observed only in the case of fusion with chaperone Caf1M and maltose-binding protein MBP expressed as precursors with their own signal sequences. The study of the isolated MBP-ESR protein in micelles and proteoliposomes demonstrated formation and decay of the main photocycle intermediates at pH > 8. The photoelectric response of the fusion proteins Caf-ESR and MBP-ESR is comparable in amplitude to the wild-type ESR response, indicating their homogeneous orientation in the proteoliposome membrane. Conclusions: The obtained constructs can be used to create bacterial expression systems for various retinal proteins, ensuring their uniform incorporation into proteoliposomes. [ABSTRACT FROM AUTHOR]

Post-harvest decay of fresh agricultural produce is a major threat to food security globally. Synthetic fungicides, commonly used in practice for managing the post-harvest losses, have negative impacts on consumers' health. Studies have reported the effectiveness of fungal isolates from plants as biocontrol agents of post-harvest diseases, although this is still poorly established in tomatoes (Solanum lycopersicum L. cv. Jasmine). In this study, 800 endophytic fungi were isolated from mature green and ripe untreated and fungicide-treated tomato fruits grown in open soil and hydroponics systems. Of these, five isolates (Aureobasidium pullulans SUG4.1, Coprinellus micaceus SUG4.3, Epicoccum nigrum SGT8.6, Fusarium oxysporum HTR8.4, Preussia africana SUG3.1) showed antagonistic properties against selected post-harvest pathogens of tomatoes (Alternaria alternata , Fusarium solani , Fusarium oxysporum , Geotrichum candidum , Rhizopus stolonifera, Rhizoctonia solani), with Lactiplantibacillus plantarum as a positive control. P. africana SUG3.1 and C. micaceus SUG4.3 significantly inhibited growth of all the pathogens, with antagonistic capabilities comparable to that exhibited by L. plantarum. Furthermore, the isolates produced an array of enzymes, including among others, amylase, cellulose and protease; and were able to utilize several carbohydrates (glucose, lactose, maltose, mannitol, sucrose). In conclusion, P. africana SUG3.1 and C. micaceus SUG4.3 may complement L. plantarum as biocontrol agents against post-harvest pathogens of tomatoes. • 800 endophytic fungi were isolated from mature green and ripe tomatoes. • Some exhibited antagonistic activity against post-harvest pathogens of tomatoes. • Preussia africana SUG3.1 and Coprinellus micaceus SUG4.4 suppress post-harvest fungal infections. • Antibiosis and competition for nutrients appears to be the main mechanism of action. [ABSTRACT FROM AUTHOR]

Soluble sugar is one of the important characteristic metabolites contributing to the flavor quality of tea. However, little is known about the changes and metabolic pathways of sugar in summer–autumn black tea varieties during the solid-state fermentation of Ganoderma lucidum. Therefore, this study is aimed at investigating the effects of the solid-state fermentation of Ganoderma lucidum on soluble sugar metabolism in three summer–autumn black tea varieties. In this study, we combined targeted metabolomics technology, gas chromatography-mass spectrometry, and multivariate statistical analysis to reveal the metabolic rules of 13 different sugars in three varieties of summer–autumn black tea under the action of Ganoderma lucidum. Using multivariate statistical analysis, 8 common key sugar differential metabolites were selected from the three groups of samples: D-fructose, trehalose, lactose, maltose, D-galactose, glucose, L-rhamnose, and xylitol. The eight different sugars contributed to the quality difference between Ganoderma lucidum tea and black tea in this study. To better explore the patterns of sugar changes, the metabolic pathways of 13 different sugars were screened and analyzed. The result revealed that the solid-state fermentation of Ganoderma lucidum increased the sweet characteristics and decreased the bitter characteristics of the three summer–autumn black tea varieties. In addition, these results showed that the solid-state fermentation of Ganoderma lucidum could produce lactose in tea, reduce glucose and fructose contents, and increase xylitol content. The solid-state fermentation of Ganoderma lucidum can regulate soluble sugar metabolism in three summer–autumn black tea varieties, leading to an improvement in their quality. This study has potential practical significance for regulating the flavor quality of summer–autumn tea. [ABSTRACT FROM AUTHOR]

Cold stress severely restricts plant development, causing significant agricultural losses. We found a critical transcription factor network in Medicago ruthenica was involved in plant adaptation to low‐temperature. APETALA2/ethylene responsive factor (AP2/ERF) transcription factor MrERF039 was transcriptionally induced by cold stress in M. ruthenica. Overexpression of MrERF039 significantly increased the glucose and maltose content, thereby improving the tolerance of M. ruthenica. MrERF039 could bind to the DRE cis‐acting element in the MrCAS15A promoter. Additionally, the methyl group of the 14th amino acid in MrERF039 was required for binding. Transcriptome analysis showed that MrERF039 acted as a sugar molecular switch, regulating numerous sugar transporters and sugar metabolism‐related genes. In addition, we found that MrERF039 could directly regulate β‐amylase gene, UDP glycosyltransferase gene, and C2H2 zinc finger protein gene expression. In conclusion, these findings suggest that high expression of MrERF039 can significantly improve the cold tolerance of M. ruthenica root tissues during cold acclimation. Our results provide a new theoretical basis and candidate genes for breeding new legume forage varieties with high resistance. Summary statement: MrERF039 plays a positive role in the cold response of Medicago ruthenica by directly regulating sugar‐related genes, including β‐amylase through direct binding to DRE cis‐acting elements, increasing glucose and maltose content, and consequently improving the cold tolerance of Medicago ruthenica. [ABSTRACT FROM AUTHOR]

Intelectins belong to a family of lectins with specific and transitory carbohydrate interaction capabilities. These interactions are related to the activity of agglutinating pathogens, as intelectins play a significant role in immunity. Despite the prominent immune defense function of intelectins, limited information about its structural characteristics and carbohydrate interaction properties is available. This study investigated an intelectin transcript identified in RNA-seq data obtained from the South American lungfish (Lepidosiren paradoxa), namely LpITLN2-B. The structural analyses predicted LpITLN2-B to be a homo-trimeric globular protein with the fibrinogen-like functional domain (FReD), exhibiting a molecular mass of 57 kDa. The quaternary structure is subdivided into three monomers, A, B, and C, and each domain comprises 11 β-sheets: an anti-parallel β-sheet, a β-hairpin, and a disordered β-sheet structure. Molecular docking demonstrates a significant interaction with disaccharides rather than monosaccharides. The preferential interaction with disaccharides highlights the potential interaction with pathogen molecules, such as LPS and Poly(I:C). The hemagglutination assay inhibited lectins activity, especially maltose and sucrose, highlighting lectin activity in L. paradoxa samples. Overall, our results show the potential relevance of LpITLN2-B in L. paradoxa immune defense against pathogens. [ABSTRACT FROM AUTHOR]

Context: Further understanding of the molecular mechanisms is necessary since it is important for designing new drugs. This study aimed to understand the molecular mechanisms involved in the design of drugs that are inhibitors of the α-glucosidase enzyme. This research aims to gain further understanding of the molecular mechanisms underlying antidiabetic drug design. The molecular docking process yielded 4 compounds with the best affinity energy, including γ-Mangostin, 1,6-dimethyl-ester-3-isomangostin, 1,3,6-trimethyl-ester-α-mangostin, and 3,6,7-trimethyl-ester-γ-mangostin. Free energy calculation with molecular mechanics with generalized born and surface area solvation indicated that the 3,6,7-trimethyl-γ-mangostin had a better free energy value compared to acarbose and simulated maltose together with 3,6,7-trimethyl-γ-mangostin compound. Based on the analysis of electrostatic, van der Waals, and intermolecular hydrogen interactions, 3,6,7-trimethyl-γ-mangostin adopts a noncompetitive inhibition mechanism, whereas acarbose adopts a competitive inhibition mechanism. Consequently, 3,6,7-trimethyl-ester-γ-mangostin, which is a derivative of γ-mangostin, can provide better activity in silico with molecular docking approaches and molecular dynamics simulations. Method: This research commenced with retrieving protein structures from the RCSB database, generating the formation of ligands using the ChemDraw Professional software, conducting molecular docking with the Autodock Vina software, and performing molecular dynamics simulations using the Amber software, along with the evaluation of RMSD values and intermolecular hydrogen bonds. Free energy, electrostatic interactions, and Van der Waals interaction were calculated using MM/GBSA. Acarbose, used as a positive control, and maltose are simulated together with test compound that has the best free energy. The forcefields used for molecular dynamics simulations are ff19SB, gaff2, and tip3p. [ABSTRACT FROM AUTHOR]

In order to improve the extraction rate of active components in fig leaves, the fermentation of fig leaves by bacterial enzyme was studied. A strain of Stenotrophomonas maltophilia was isolated from fresh, frozen, and dried fig leaves using fig leaves as the sole carbon source. It was found that the coumarin content of fermented fig leaves was higher than that of unfermented fig leaves. Therefore, this bacterium strain was selected as the fermentation strain in this experiment. After fermentation for two days, the coumarin and terpenes contents in the fermentation broth were determined by colorimetry, and the culture conditions were optimized by single factor and orthogonal experiments. The results showed that the optimal fermentation mode was bacterial-enzyme co-fermentation, the optimal precursor was 8 mg/L of L-phenylalanine, the additional carbon source was 2 g/L maltose, the optimum nitrogen source was ammonium sulfate + peptone mixture of 3 g/L, and the optimum ratio of cellulase and pectinase was 1: 2. At the optimization condition, the contents of coumarins, terpenoids, flavonoids, and polysaccharides in the fermentation broth of fig leaves were increased by 92.66%, 59.40%, 44.68%, and 42.84% respectively. The content of polyphenols was 4.24 times higher than that before optimization. The study indicates that fig leaves fermentation can increase the content of its active components, and the study provides a basis for the development and utilization of fig leaves as a new feed additive. [ABSTRACT FROM AUTHOR]

• The most prominent changes in pea (Pisum sativum L.) of sugars occurred within 96 h after germination. • Nano-selenium application promotes starch degradation, inhibit crude fiber synthesis, and a significant increase in total soluble sugar content. • 56 differentially expressed genes are screened related to sugar metabolism by transcriptome analysis of pea sprouts germinated by nano-selenium. • Hexokinase (HK) gene has the highest relative expression in the tissues of pea sprouts verified qRT-PCR. • Selenium-enriched pea sprouts are valuable as innovative health foods. Selenium, a beneficial trace element, plays a vital role in plant growth. In this study, the effects of nano-selenium on pea (Pisum sativum L.) germination were characterized. Nano-selenium application led to decreases in starch, crude fiber, and soluble polysaccharide contents, but a significant increase in total soluble sugar content. The most prominent changes in sugars occurred 96 h after germination commenced. The nano-selenium treatment significantly increased glucose content in roots, slightly decreased fructose content in the hypocotyl, and increased fructose content in roots and seed. Oligosaccharide content was higher in seed than in the hypocotyl and roots, with sucrose being the highest, followed by maltose. The transcriptome analysis of pea tissues at different stages of germination detected 56 differentially expressed genes related to sugar metabolism. The verification of expression of five selected key genes by qRT-PCR showed the hexokinase (HK) gene had the highest relative expression in the tissues of pea sprouts. There were significant differences in gene expression in the pea sprouts cultured with nano-selenium compared with the control group. Nano-selenium promoted the expression of genes associated with sugar metabolism in roots and seeds, promoted the synthesis of soluble sugars in the sprouts, accelerated the amylolysis, and inhibited cellulose synthesis. [ABSTRACT FROM AUTHOR]

Simple Summary: Different nutritional supplements can differentially affect the reproductive performance of parasitoid wasps, and selection of optimal diets can increase parasitoid quality, increasing their effectiveness as biological control agents. Binodoxys communis (Gahan) (Hymenoptera: Aphelinidae) is the main parasitoid attacking Aphis gossypii Glover (Hemiptera: Aphidae) in Xinjiang, China. In this study we tested the effects of fructose, glucose, sucrose, trehalose, maltose, melezitose, and sorbitol on the longevity, parasitism ability, parasitism behavior, and flight ability of B. communis. We found that, compared with other sugars, B. communis fed on glucose, sucrose, or fructose had significantly greatly longevity and parasitism ability. Also, parasitoids that were fed these sugars interacted with A. gossypii more frequently during parasitization and had an enhanced ability. The above information helps in the evaluation of nutritional supplements, both to improve the parasitoid's reproductive performance and its effectiveness as a biological control agent. Alterative nutritional foods consumed by adult parasitoids play an important role in their fitness and ability to control pests because of food scarcity in many crops. While adult parasitoids feed on various sugars, they vary in their nutritional value for parasitoids. We assessed the effects of seven sugars (fructose, glucose, sucrose, trehalose, maltose, melezitose, and sorbitol) on the longevity, parasitism ability, parasitism behavior, and flight ability of B. communis, an important parasitoid of cotton aphids. We found that access to glucose, sucrose, or fructose, increased B. communis adult longevity more than the other sugars offered. All sugars except trehalose increased the parasitism rate to more than 50% compared to the starved control (only provided with water). We then compared parasitoid behaviors of wasps fed glucose, sucrose, or fructose to that of the starved control (with access only to water) and found that those fed B. communis spent more time either examining or attacking aphids than parasitoids in the control group, which spent more time walking or resting. Also, consumption of glucose, sucrose, or fructose also significantly improved the flight ability (the total flight distance, flight time, and average flight speed) of B. communis. [ABSTRACT FROM AUTHOR]

In wine, lactic acid bacteria (LAB) are responsible for the bioconversion of malic acid to lactic acid, malolactic fermentation that mainly aims at reducing wine acidity. Two LAB strains isolated from the red wine microbiota (Oenococcus oeni 13-7 and Lactobacillus plantarum R1-1), were tested for their ability to exhibit the carbon catabolite repression (CCR) mechanism, that allows the rapid use of certain carbohydrates, over other carbon sources. Bacterial cells were inoculated in 0.1 M glycine buffer (pH 3.5), incubated at 30°C, with different carbohydrates (45 mM) and malic acid (45 mM). For both strains, the presence of glucose significantly inhibited malic acid metabolization (-60%), a similar effect being observed for galactose, mannose and maltose. The highest rate of malic acid conversion was shown in fructose/malate medium. Obtained results showed that malolactic strains can control the utilization of carbon sources via CCR, further studies being necessary to elucidate the mechanisms underlying this process. [ABSTRACT FROM AUTHOR]

Puerarin 6′′-O-acetate (P6′′A), a puerarin derivative with a high stability and an improved permeability, exhibits a promising potential in drug development. The large-scale production of P6′′A through green synthesis procedures has never been achieved owing to the low availability of acetyl-CoA. Herein, the enzymatic synthesis of P6′′A in Escherichia coli cell factories overproducing acetyl-CoA has been achieved. Firstly, the effect of three acetyl-CoA-producing pathways on P6′′A conversion in E.coli was compared comprehensively. The pathways containing acetyl-CoA synthetase (ACS) or the pyruvate dehydrogenase (PDH) complex were thereby identified as favorable fluxes yielding more acetyl-CoA. Hence, cells co-expressing ACS and the PDH complex were selected as cell factories overproducing acetyl-CoA owing to the positive cooperativity of ACS and PDH. Subsequently, the maa gene encoding maltose acetyltransferase (MAT) was introduced into these acetyl-CoA-producing cell factories to form engineered strains Q1–Q14. After condition optimization, the engineered strain Q5 got a P6′′A yield of 0.98 g L−1 in M9P-Glc medium supplemented with 2.5 mM puerarin and 1.5% glucose. To further improve P6′′A production, the maa gene in Q5 was replaced by a mutant maaE125N gene to generate another engineered strain (Q15). Microbial production of P6′′A in Q15 was achieved through the centrifuge tube culture, the shake-flask culture and the 6-L reactor culture and the highest P6′′A yield of up to 2.02 g L−1 was obtained in the reactor fermentation. Enzymatically synthesized P6′′A displayed a high stability in human and mouse plasma, a dose-dependent cytotoxicity in HepG2 cells and an improved liposolubility. This research provides a feasible procedure for the green synthesis of puerarin monoesters and lays a foundation for druggability research on P6′′A. [ABSTRACT FROM AUTHOR]

Background: Chalkiness is a common phenotype induced by various reasons, such as abiotic stress or the imbalance of starch synthesis and metabolism during the development period. However, the reason mainly for one gene losing its function such as NAC (TFs has a large family in rice) which may cause premature is rarely known to us. Results: The Ko-Osnac02 mutant demonstrated an obviously early maturation stage compared to the wild type (WT) with 15 days earlier. The result showed that the mature endosperm of Ko-Osnac02 mutant exhibited chalkiness, characterized by white-core and white-belly in mature endosperm. As grain filling rate is a crucial factor in determining the yield and quality of rice (Oryza sativa, ssp. japonica), it's significant that mutant has a lower amylose content (AC) and higher soluble sugar content in the mature endosperm. Interestingly among the top DEGs in the RNA sequencing of N2 (3DAP) and WT seeds revealed that the OsBAM2 (LOC_Os10g32810) expressed significantly high in N2 mutant, which involved in Maltose up-regulated by the starch degradation. As Prediction of Protein interaction showed in the chalky endosperm formation in N2 seeds (3 DAP), seven genes were expressed at a lower-level which should be verified by a heatmap diagrams based on DEGs of N2 versus WT. The Tubulin genes controlling cell cycle are downregulated together with the MCM family genes MCM4 (↓), MCM7 (↑), which may cause white-core in the early endosperm development. In conclusion, the developing period drastically decreased in the Ko-Osnac02 mutants, which might cause the chalkiness in seeds during the early endosperm development. Conclusions: The gene OsNAC02 which controls a great genetic co-network for cell cycle regulation in early development, and KO-Osnac02 mutant shows prematurity and white-core in endosperm. [ABSTRACT FROM AUTHOR]

Bioethanol is an organic compound resulted from the fermentation of sugar substrates by microorganisms which is used as alternative energy sources. During bioethanol fermentation yeast are exposed to various fermentation stresses, including temperature, osmotic, and oxidative stresess. Such conditions may decrease ethanol production. We previously isolated fermentation-stress tolerance yeast isolates from traditional Balinese beverages, identified as Wickerhamomyces anomalus BT2, BT5, and BT6. However no data available regarding the bioethanol production of those isolates. Our study indicates that these strains could utilize various sugar substrates (glucose, xylose, maltose, sucrose) in oxidative fermentative media. The highest value of substrate utilization efficiency following 48 hours fermentation was shown by BT6 on glucose (61.02%), BT 2 on xylose (55.44%) and maltose (60.90%). Measurement of ethanol production by Gas Chromatography showed that the strains were able to produce higher ethanol on the glucose substrate than other substrates. For instance, BT6 could produce the highest ethanol production (5.00 g/L) amongst strains tested by using glucose as substrate. Yet, the particular strains could only produce 0.30 g/L and 0.65 g/L by using xylose and maltose, respectively. For further genetic engineering purposes, we detected ADH1 gene from all three isolates, with high homology to the alcohol dehydrogenase from Saccharomyces cerevisiae, Geobacillus stearothermophilus and Pseudomonas aeruginosa. Further strain development can be carried out targeting the ADH1 gene, important for ethanol fermentation. [ABSTRACT FROM AUTHOR]

Bee pollen is a well-balanced food and food supplement whose nutritional and bioactive properties vary according to its botanical origin. This comparative study examined some nutritional and biochemical properties of the various types of bee pollen samples collected from Anatolian region. Eleven samples were examined based on melissopalynological analyses, including five unifloral types (chestnut, rhododendron, buckwheat, ivy, and rose), and six different multifloral samples. Crude protein, edible fiber, sugar (fructose, glucose, sucrose, maltose), and essential elements (Na, K. Ca, Mg, Fe, Cu, Zn, and Mn) were evaluated for chemical characterization. Twenty-five phenolic components were analyzed using reverse-phase high-performance liquid chromatography (RP-HPLC–PDA). Total phenolic compounds, total flavonoids, ferric reducing antioxidant capacity (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity were measured as antioxidant determinants. The results were subjected to statistical analysis, and principal component analysis (PCA) was applied to determine differences between pollen samples. Chrysin, t-cinnamic acid, and pinocembrin were identified as common components in all samples, but in varying amounts. Bee pollens' nutrients and phenolic compositions, thus, varied according to their botanical properties, although chestnut pollen exhibited the greatest antioxidant capacity. [ABSTRACT FROM AUTHOR]

Glucagon, a peptide hormone clinically used to treat acute hypoglycemia in diabetes patients, is readily degenerated by undergoing fibrillation under pharmaceutical conditions. Since glucagon is employed as a typical model system for structural investigation of amyloid fibril formation of proteins, production of recombinant glucagon is in demand to facilitate mutagenesis and isotope labeling for nuclear magnetic resonance (NMR) studies. In this study, we established a method to produce recombinant glucagon in Escherichia coli. Recombinant plasmids were constructed to express a maltose-binding protein-fused glucagon, which was cleaved by factor-Xa protease to yield glucagon peptide without any N-terminal extra residues. The final product was clearly identified and characterized by immunoblotting, mass spectrometry, circular dichroism spectroscopy, and backbone NMR assignments of the [13C/15N] isotope-enriched sample. Cellular activity of the recombinant glucagon in hepatocytes was confirmed by monitoring characteristic gene expressions. Site-directed mutagenesis was successfully performed using our recombinant production system. Our bacterial production system and the recombinant glucagon not only provide an economical route of glucagon manufacturing, but also enable NMR-based structural investigation of glucagon fibrillation. [Display omitted] • Bacterial production of a recombinant glucagon was established using Escherichia coli. • MBP used as a fusion tag was cleaved by factor-Xa to yield isolated glucagon. • Final product was validated by mass spectrometry and activity assay in hepatocytes. • Mutagenesis and isotope labeling were successful using the production system. • Backbone NMR assignments were completed by heteronuclear multidimensional NMR. [ABSTRACT FROM AUTHOR]

Using sugar molecules as lyophilization protectants in HSA nanocolloid kit formulations is critical to obtaining kits with excellent stability. A thermal gelation method with no sugar treatment and variations in sugar (glucose, maltose, and lactose) was applied to produce these kits. The particle size and radiochemical purity results showed that HSA nanocolloid kits with sugar excipients are more stable than those without sugar. The addition of lactose as an excipient (HSA: lactose 1:10) results in a stable nanocolloid system, allowing for the development of novel lyophilized HSA nanocolloid kit formulations. [ABSTRACT FROM AUTHOR]

The kefir‐based smoothies with kale and spinach were designed as a ready‐to‐drink and innovative functional snack. Microbiological, physicochemical, as well as pre‐ and postgastrointestinal total antioxidant capacity (TAC; CUPRAC, DPPH, and FRAP) analyses were conducted. It was determined that the kefir‐based smoothies with vegetables had higher ash, carbohydrate, and dietary fiber values. Fructose and glucose contents of smoothy with kale were high, while smoothy with spinach included high sucrose and maltose. The microbiology results revealed that kefir‐based vegetable smoothies had minimum Lactobacillaceae viability (>log 7 cfu g−1) for the required functional effect after 14‐day storage. Moreover, the addition of kale significantly increased (p <.01) the level of initial TAC (CUPRAC, DPPH, and FRAP) and total phenolic content (TPC) values. After in vitro gastric digestion analysis, smoothie with spinach demonstrated higher TAC and TPC values and the control sample had higher TAC and TPC values compared with a predigestion step. It was found that in vitro intestinal DPPH values were higher for the sample with spinach samples, while the sample with kale had the highest FRAP values. It was also found that the bioaccessibility indexes of plain kefir were determined to be the highest in both in vitro gastric and intestinal procedures. The present study provided novel insights into the in vitro digestion properties of kefir fortified with vegetables. Nevertheless, further studies are needed to identify the functional properties of the milk and plant matrices mixture using in vitro and in vivo trials. [ABSTRACT FROM AUTHOR]

The acute toxicity study is a crucial component of preclinical toxicology assessments conducted to evaluate the potential adverse effects of a substance or compound on albino rabbits. This study provides key findings and implications of an acute toxicity study of natural and synthetic polymers that are used in the preparation of transdermal microneedle patches (TMNPs) for sustained delivery of doxazocin mesylate. In this study, we divided the albino rabbits into three equal groups (n = 6). Group-I was labeled as a control group and group-II was treated with TMNPs-1 (HPMC, maltose) and group-III was treated with TMNPs-2 (PLGA, polyvinyl acetate). The results obtained from feed consumption, hematology profile, biochemical studies, vital organ weight, and histopathological changes in vital organs were compared with the control group, and proved the safety profile of the polymers involved in the fabrication of microneedle patches. Mechanical strength and dermal safety study established the facts about the safety of microneedle patches and potential risks to health as a carrier system for the delivery of drugs into the systemic circulation directly. [ABSTRACT FROM AUTHOR]

Disaccharides are composed of two monosaccharide subunits joined by a glycosidic linkage in an α or β configuration. Different combinations of isomeric monosaccharide subunits and different glycosidic linkages result in different isomeric disaccharide products. Thus, direct discrimination of these disaccharide isomers from a mixture is extremely difficult. In this paper, a hetero‐octameric Mycobacterium smegmatis porin A (MspA) nanopore conjugated with a phenylboronic acid (PBA) adapter was applied for disaccharide sensing, with which three most widely known disaccharides in nature, including sucrose, lactose and maltose, were clearly discriminated. Besides, all six isomeric α‐D‐glucopyranosyl‐D‐fructoses, differing only in their glycosidic linkages, were also well resolved. Assisted by a custom machine learning algorithm, a 0.99 discrimination accuracy is achieved. Nanopore discrimination of disaccharide isomers with different glycosidic linkages, which has never been previously demonstrated, is inspiring for nanopore saccharide sequencing. This sensing capacity was also applied in direct identification of isomaltulose additives in a commercial sucrose‐free yogurt, from which isomaltulose, lactose and L‐lactic acid were simultaneously detected. [ABSTRACT FROM AUTHOR]

Disaccharides are composed of two monosaccharide subunits joined by a glycosidic linkage in an α or β configuration. Different combinations of isomeric monosaccharide subunits and different glycosidic linkages result in different isomeric disaccharide products. Thus, direct discrimination of these disaccharide isomers from a mixture is extremely difficult. In this paper, a hetero‐octameric Mycobacterium smegmatis porin A (MspA) nanopore conjugated with a phenylboronic acid (PBA) adapter was applied for disaccharide sensing, with which three most widely known disaccharides in nature, including sucrose, lactose and maltose, were clearly discriminated. Besides, all six isomeric α‐D‐glucopyranosyl‐D‐fructoses, differing only in their glycosidic linkages, were also well resolved. Assisted by a custom machine learning algorithm, a 0.99 discrimination accuracy is achieved. Nanopore discrimination of disaccharide isomers with different glycosidic linkages, which has never been previously demonstrated, is inspiring for nanopore saccharide sequencing. This sensing capacity was also applied in direct identification of isomaltulose additives in a commercial sucrose‐free yogurt, from which isomaltulose, lactose and L‐lactic acid were simultaneously detected. [ABSTRACT FROM AUTHOR]

Dextranases are hydrolases that exclusively catalyze the disruption of α−1,6 glycosidic bonds. A series of variant enzymes were obtained by comparing the sequences of dextranases from different sources and introducing sequence substitutions. A correlation was found between the number of amino acids in the 397–401 region and the hydrolytic process. When there were no more than 5 amino acids in the 397–401 region, the enzyme first hydrolyzed the dextran T70 to a low molecular weight dextran with a molecular weight of about 5000, then IMOs 1 1 IMOs: isomaltooligosaccharides. G: glucose, G2: maltose, G3: maltotriose, G4: maltotetraose, G5: maltopentaose, G6: maltohexaose, G7: maltoheptaose, IG2: isomaltose, IG3: isomaltotriose, IG4: isomaltotetraose, IG5: isomaltopentaose appeared in the system if the degradation continued, showing a clear sequential relationship. And when there are more than 5 amino acids in the 397–401 region, IMOs were produced at the beginning of hydrolysis and continue to increase throughout the hydrolytic process. At the same time, we investigated the enzymatic properties of the variants and found that the hydrolytic rate of A-Ca was 11 times higher than that of the original enzyme. The proportion of IMOs produced by A-Ca was 80.68%, which was nearly10% higher than the original enzyme, providing a new enzyme for the industrial preparation of IMOs. • One of the factors affecting the hydrolysis process of Aodex was determined. • Using sequence substitution method for molecular modification. • The hydrolysis rate of variant A-Ca increased by 11 times. • The proportion of IMOs produced by A-Ca was 80.68% which increased by nearly 10% compared to the WT. [ABSTRACT FROM AUTHOR]

The present study examines the impact of nitrogen sources (yeast extract, ammonium sulfate peptone, ammonium nitrate, urea, and sodium nitrate), salt solution (0.5 g/L MgSO4, 0.5 g/L KH2PO4, 0.3 g/L CaCl2), trace elements solution (0.1 g/L CuSO4, 0.1 g/L FeSO4, 0.02 g/L MnCl2, 0.02 g/L ZnSO4), operational parameters (temperature, aeration, agitation, initial pH and xylose concentration) and co- substrate supplementation (glucose, fructose, maltose, sucrose, and glycerol) on xylitol biosynthesis by Candida tropicalis ATCC 13803 using synthetic xylose. The significant medium components were identified using the Plackett Burman design followed by central composite designs to obtain the optimal concentration for the critical medium components in shaker flasks. Subsequently, the effect of operational parameters was examined using the One Factor At a Time method, followed by the impact of five co-substrates on xylitol biosynthesis in a 1 L bioreactor. The optimal media components and process parameters are as follows: peptone: 12.68 g/L, yeast extract: 6.62 g/L, salt solution (0.5 g/L MgSO4, 0.5 g/L KH2PO4, and 0.3 g/L CaCl2): 1.23 X (0.62 g/L, 0.62 g/L, and 0.37 g/L respectively), temperature: 30 °C, pH: 6, agitation: 400 rpm, aeration: 1 vvm, and xylose: 50 g/L. Optimization studies resulted in xylitol yield and productivity of 0.71 ± 0.004 g/g and 1.48 ± 0.018 g/L/h, respectively. Glycerol supplementation (2 g/L) further improved xylitol yield (0.83 ± 0.009 g/g) and productivity (1.87 ± 0.020 g/L/h) by 1.66 and 3.12 folds, respectively, higher than the unoptimized conditions thus exhibiting the potential of C. tropicalis ATCC 13803 being used for commercial xylitol production. [ABSTRACT FROM AUTHOR]

The growing customer demand for diversified brewery products is pushing the brewery industry to search for brewer's yeasts. The aim of this study was to screen and evaluate the brewing potential of eight S. cerevisiae strains isolated from Ethiopian traditional fermented beverages. They were screened for potential brewer's yeast characteristics, and then their brewing performance were evaluated on a laboratory scale. Screening data indicated that all strains demonstrated reasonable brewing characteristics. Brewing performances evaluation has shown that tested strains showed reasonable performance. They yield ethanol content ranging between 5.2 ± 0.2 and 5.5 ± 0.2 (v/v%) with degree of attenuation varying between 64.1 ± 6.8 and 72.6 ± 2.0%. Compared to the investigated strains, the commercial strain showed remarkable performance with ethanol production (6.1 ± 0.2, v/v%) and attenuation degree (82.2 ± 2.0%). Produced beers were analyzed for sensorial attributes, and they were not statistically significant in all tested sensorial profiles, except beers produced by MABD1 and Sc. JD9, which were recognized as extremely bitter by two tasters. FT-IR spectra matching of experimental and reference beers revealed that beer samples exhibited the same spectral pattern, implying they have a similar chemical profile. Studies focused on the understanding of the determinant abiotic factors for an acceptable performance of the investigated S. cerevisiae strains at the simulated industrial-scale brewing condition is recommended. Moreover, adaptation to maltose and malto-dextrin assimilation at low temperature condition is recommended. [ABSTRACT FROM AUTHOR]

The cultivation of sweet chestnuts in Slovakia has a century-year-old tradition. However, just a few research publications provide more comprehensive information about the chestnuts' chemical composition of different varieties in individual growing areas. Sweet chestnuts contain various nutrients, of which carbohydrates are very important for assessing their commercial quality. Simple carbohydrates are mainly represented by glucose, fructose, sucrose, and maltose. The content of glucose was 0.178–3.152, fructose 0.271–3.170, sucrose 9.355–29.890, and maltose 0.171–1.584 g/100 g DW in chestnut samples from 27 sampling points, which are part of Slovakia's four largest growing areas of sweet chestnuts. Carbohydrate contents were determined by the high-performance liquid chromatography (HPLC-RI) method. There are statistically proven differences in the content of individual carbohydrates depending on the sampling area of chestnuts. Potassium has a significant presence in sweet chestnuts as a mineral substance. Its content was determined in the 377–789 mg/100 g DW range. The content of sodium (calcium, magnesium, and phosphorus) was 0.65–6.90 (26.9–103, 58.7–101, and 96.5–179) mg/100 g DW. The contents of mineral substances were determined by the AAS method. The content of carbohydrates and mineral substances is mainly influenced by variety and age. [ABSTRACT FROM AUTHOR]

Mammalian F-ATP synthase is central to mitochondrial bioenergetics and is present in the inner mitochondrial membrane in a dynamic oligomeric state of higher oligomers, tetramers, dimers, and monomers. In vitro investigations of mammalian F-ATP synthase are often limited by the ability to purify the oligomeric forms present in vivo at a quantity, stability, and purity that meets the demand of the planned experiment. We developed a purification approach for the isolation of bovine F-ATP synthase from heart muscle mitochondria that uses a combination of buffer conditions favoring inhibitor factor 1 binding and sucrose density gradient ultra-centrifugation to yield stable complexes at high purity in the milligram range. By tuning the glyco-diosgenin to lauryl maltose neopentyl glycol ratio in a final gradient, fractions that are either enriched in tetrameric or monomeric F-ATP synthase can be obtained. It is expected that this large-scale column-free purification strategy broadens the spectrum of in vitro investigation on mammalian F-ATP synthase. [ABSTRACT FROM AUTHOR]

We investigated the powderization of flavoring substances, using an amorphous solid dispersion (ASD) technique, in which hydrophobic molecules are separately embedded in a water-soluble carrier matrix. Six flavors, five carrier forming materials (polyvinylpyrrolidone/disaccharides), two solvents (methanol/ethanol) and two drying methods (vacuum-foam-/spray-drying) were employed. The drying conditions for the two drying processes were first examined, and under the optimal drying conditions, various flavor-carrier combinations and compositions of ASD samples were prepared and their flavor retention after drying and during storage under a vacuum were compared. Results demonstrated that flavor loss during drying and storage was minimized when the material was vacuum-foam-dried with polyvinylpyrrolidone. Vacuum-foam-drying in the presence of α-maltose or palatinose also resulted in a greater retention of flavor during drying and storage than a typical O/W emulsification-based powderization. These findings suggest that the ASD-based powderization of flavoring materials is a feasible alternative to the currently used produces. [ABSTRACT FROM AUTHOR]

Though industrial progress has improved mankind's standard of living, it has also degraded the environment. The lack of freshwater in the cosmos has led researchers to pay close attention to water contamination. The holmium silicate nanostructures with Ho 2 Si 6 O 9 formula were prepared through a multifuel auto-combustion process involving starch, fructose, and maltose. Obtaining Ho 2 Si 6 O 9 in the presence of starch results in a pure and morphologically desirable sample. Considering the suitable bandgap (3.01 eV) of Ho 2 Si 6 O 9 , this compound could be a good candidate for the degradation of toxic organic compounds in the UV range. Several factors play a crucial role in the purification of water, such as the type of organic contamination, the content of catalysts, and the concentration of pollutants. Results showed that 50 mg of Ho 2 Si 6 O 9 decomposed 5 ppm thymol blue in a high percentage (91.4%). The scavenger test revealed that superoxide (•O 2 −) species played a significant role in photodegradation. The kinetics study confirmed that the maximum efficiency has a maximum rate constant (k = 0.0191 min−1). [Display omitted] • Fabrication of Nano-Ho 2 Si 6 O 9 by a facile and affordable auto-combustion method. • Different fuels affected the morphology and size of the product. • Nominating Ho 2 Si 6 O 9 as a potential candidate for photocatalytic water treatment. • Degradation of 91.4% thymol blue by Ho 2 Si 6 O 9 nanostructures. • •O 2 − radical performed the major role in the degradation of thymol blue. [ABSTRACT FROM AUTHOR]

Novosphingobium species are Gram-negative, rod-shaped bacteria containing sphingoglycolipids. In this study, Novosphingobium species were isolated and characterized based on morphology and biochemical analysis. The research also investigated the impact of different culture media, temperature, time and pH on the growth of Novosphingobium species, aiming to determine its optimal growth kinetics. The culture media (LB, 0.1% glucose LB, NB, 0.1% glucose NB, and MM), pH (4, 5, 6, 7, 8, 9 and 10), and incubation temperatures (25, 30, 37, 45, and 55℃), were examined for representative Novosphingobium sp. growth, measured as optical density (OD). The results indicated that all the strains displayed yellow colony morphology with rounded raised edges and smooth surfaces. Strains were rod-shaped without spore production. All the strains assimilated maltose, sucrose, xylose and oxidized glucose. Representative strain of Novosphingobium species grown rapidly on 0.1% glucose LB medium and optimal temperature were identified 37℃. The growth of representative strain was significantly influenced by the initial pH of the culture media, but 7 pH was identified as optimal. This study provides valuable information regarding the characterization and optimal growth conditions of Novosphingobium species. [ABSTRACT FROM AUTHOR]

Microbial species often develop on the roots of plants, which have a positive influence on their growth and susceptibility to diseases, among which the gram-positive rhizobacterium Bacillus subtilis stands out. The ability to form surface-attached microbial communities called biofilms is critical for the ability of B. subtilis to adhere to and protect plant roots. In this study, we demonstrated that monosaccharides, that are the main structure moiety of plant polysaccharides, play an important role in biofilm formation and plant attachment of B. subtilis during their interaction with E. coli K-12 strains. Effects of the content of maltose, xylose, lactose, glucose and saccharose in the cultural medium on bacterial biofilm formation and the mechanisms were analysed by the crystal violet staining method combined with cultivated microbial analysis, and confocal laser scanning microscopy. With the most pronounced inducing effect on the process of formation of biofilms formed as a result of the co-cultivation of B. subtilis 170 and E. coli K-12 1655, B. subtilis 168 and E. coli K-12 1655 strains, distinguishes glucose among the tested carbohydrates, but the values of their optical density, average thickness, relative spreading area, and area-to-volume ratio depend on their type in the culture medium. The content of glucose, lactose, sucrose and maltose in the culture medium determined the competitive relationships between B. subtilis 170 and E. coli K-12 1655, B. subtilis 168 and E. coli K-12 1655 strains, while the presence of lactose determined the antagonistic relationships. Individual carbohydrate sources have a different impact on the number of spores in the structure of biofilms formed by the participation of B. subtilis 170 and E. coli K-12 1655, B. subtilis 168 and E. coli K-12 1655 strains. [ABSTRACT FROM AUTHOR]

This study aimed to determine the effects of polyethylene glycol (PEG) as an abiotic elicitor and nutritional factors (different ammonium/nitrate ratios, carbon source and amount) in the culture medium on the production of Caffeic Acid Derivatives (CADs) in callus cultures of Echinacea purpurea L. Petiole and root explants were cultured on MS medium modified in terms of different types (sucrose and maltose) and amounts (sucrose 15, 45, 60 g l-1, and maltose 15, 30, 45, 60 g l-1) of carbon source, different concentrations (5, 10, 15 g l-1) of PEG and ammonium nitrate ratios (0:35, 5:25, 15:15, 35:0 mM). The amounts of CADs in the callus obtained at the end of the 10-week culture period were analysed. In both explant types, the highest amount of CADs were obtained from the medium containing 15 g l-1 sucrose and 15 or 30 g l-1 maltose applications, while the highest amount of CADs was obtained in the medium containing 0:35 mM ammonium/nitrate in nitrogen applications. While the highest amount of CADs in root explant was obtained from the medium containing 10 g l-1 PEG applications, CADs content could not be obtained in petiole explant. As a result, the highest amounts of caftaric, chlorogenic, caffeic, and chicoric acids (respectively, 9.38, 0.71, 0.29, and 34.77 mg g-1) were determined at callus obtained from root explant cultured on MS medium containing 30 g l-1 sucrose and 0:35 mM ammonium/nitrate. In conclusion, optimization of culture conditions and different elicitor applications were made to increase secondary metabolite content in E. purpurea L. under in vitro conditions and the results obtained were presented comparatively. [ABSTRACT FROM AUTHOR]

This study characterized a novel intracellular α-glucosidase, produced by Debaryomyces hansenii. The enzyme was purified through ultra-filtration, ammonium sulfate precipitation, chromatography Q-Sepharose and Sephacryl S-200 column. SDS-PAGE analysis confirmed the molecular weight of the purified protein to be 125 kDa. The enzyme exhibited maximum activity at pH 4.5and 50 °C and stability at pH 4.0 – 6.0 at 30 – 60 °C. The optimum isomalto-oligosaccharides (IMOs) bio-synthesis temperature using 400 g/L maltose was evaluated to be 35 °C with an optimum enzyme unit determined as 4.34 U g−1. The enzyme exhibits maximum maltose utilization of 78%, with or without inorganic salts, and was inhibited by ZnCl 2 , CoCl 2 and CuSO 4. Using maltose, the K m and V max values were 7.55 µM and 909 µmol/ml/mg respectively. In 3 L bioreactor, 4.34 U of enzyme produced IMOs with 47% purity utilizing 400 g/L maltose at 35 °C for 14 h. Furthermore, low purity IMOs was passed through cationic ion exchange resins to obtain high purity IMOs by separating glucose and maltose. This pioneering study produced high purity IMOs with 89% purity and 60% yield with greater conversion efficiency using a purified enzyme with a higher concentration of maltose and chromatographic method, which were confirmed by HPLC. [Display omitted] • Highly active α-glucosidase from D.hansenii was homogenized and purified. • Optimization studies enhanced isomaltooligosaccharides (IMOs) purity and yield. • Production of IMOs (47% purity) through biotransformation in a 3 L bioreactor. • High purity IMOs (89%) production using chromatographic separation. • IMOs component distribution confirmed by HPLC-RID. [ABSTRACT FROM AUTHOR]

This study evaluated the energy recovery of citrus wastes with pure cultures (Clostridium acetobutylicum ATCC 824, Clostridium beijerinckii ATCC 10132 – Inocula 1 and 2, respectively) and an autochthonous microbial (Inoculum 3) from citrus wastewater (CW) in anaerobic batch reactors to produce biofuels (H 2 and ethanol) fed with different carbon sources (glucose, maltose, fructose, sucrose, xylose, lactose, starch, acetate, and glycerol) in addition to CW. 4 conditions are tested: 1st - Inoculum 3, composed by Bacillus (20.44%), Clostridium (13.65%) and Lactobacillus (10.61%) was able to produce H 2 with sucrose, glucose and fructose. 2nd - The highest metabolites were produced with TYA medium with Inocula 1 (374 mg L−1 of acetic acid) and 2 (611 mg L−1 of butanol). Inoculum 3 generated the highest ethanol (1571.8 mg L−1) in PYG medium. 3rd - In the same nutritional conditions (PYG), Inocula 1 and 2 produced H 2 (36.80 and 64.10 mmol L−1, respectively) and acetic acid (246 and 201 mg L−1, respectively); the H 2 was suppressed with inoculum 3 due to Lactobacillus genus. 4th H 2 production was favored by autochthonous bacteria in CW. This study valued the autochthonous microorganisms in the possibility of bioconversion of organic matter into value-added products in addition to contribute to an environment-sustainable solution for citrus processing. [Display omitted] • Autochthonous consortium from citrus wastewater was able to produce 37.8 mmol H 2 L−1. • Bacillus , Clostridium and Lactobacillus were identified in autochthonous microbial. • Clostridium beijerinckii ATCC 10132 was able to produce 611 mg L−1 of butanol. • Autochthonous microbial generated the highest ethanol (1571.8 mg L−1) in PYG medium. [ABSTRACT FROM AUTHOR]

Biological dihydrogen (H 2) production is a promising alternative to the conventional non-renewable energies. H 2 production by dark fermentation is particularly studied regarding its independence from fossil fuel and electricity, its cultivation into bioreactors and the broad range of substrate that can be used. A major goal application of dark fermentation is the valuation of wastes. In this context, cheese whey wastes are particularly adapted, since rich in organic matter, abundant and low cost. A novel H 2 -producing strain was isolated from a sample of mixed bovine manure and cheese whey, collected in a cheese-producing dairy farm. The strain was identified as a Clostridium beijerinckii strain based on morphological and physiological characteristics, and 16S rDNA sequencing. The optimum temperature and pH for H 2 production was 40 °C and pH 7, respectively. Substrate and stress tolerance tests showed that C. beijerinckii C.sp.1.3 could produce H 2 from glucose, lactose, maltose, cellobiose, galacturonic acid, xylose, and sucrose, and to a lesser extent from starch, cellulose and glycerol. The H 2 production potential from a mix of dairy industrial wastes of this strain has been evaluated. The highest productivity in batch for 5 g L−1 of Total Organic Carbon (TOC) was 833.1 mL H 2 L−1 d−1, with a yield of 2.03 mM H2 mM lactose −1, demonstrating the potential of this strain for H 2 production from industrial wastes. • An H 2 -producing strain was isolated from a mixture of manure and dairy waste. • The strain has the potential of fermenting a broad range of economic substrates. • Mixture of crude cheese whey and wastewater were used as model for H 2 production. • Productivity reach up to 833.1 mL L−1 d−1 with a yield of 2.03 mol H2.mol lactose −1. • A mechanism of lactate-driven fermentation was observed. [ABSTRACT FROM AUTHOR]

Cyclodextrinases are carbohydrate-active enzymes involved in the linearization of circular amylose oligosaccharides. Primarily thought to function as part of starch metabolism, there have been previous reports of bacterial cyclodextrinases also having additional enzymatic activities on linear malto-oligosaccharides. This substrate class also includes environmentally rare α-diglucosides such as kojibiose (α-1,2), nigerose (α-1,3), and isomaltose (α-1,6), all of which have valuable properties as prebiotics or low-glycemic index sweeteners. Previous genome sequencing of three Cellvibrio japonicus strains adapted to utilize these α-diglucosides identified multiple, but uncharacterized, mutations in each strain. One of the mutations identified was in the amy13E gene, which was annotated to encode a neopullulanase. In this report, we functionally characterized this gene and determined that it in fact encodes a cyclodextrinase with additional activities on α-diglucosides. Deletion analysis of amy13E found that this gene was essential for kojibiose and isomaltose metabolism in C. japonicus. Interestingly, a Δamy13E mutant was not deficient for cyclodextrin or pullulan utilization in C. japonicus; however, heterologous expression of the gene in E. coli was sufficient for cyclodextrin-dependent growth. Biochemical analyses found that CjAmy13E cleaved multiple substrates but preferred cyclodextrins and maltose, but had no activity on pullulan. Our characterization of the CjAmy13E cyclodextrinase is useful for re fining functional enzyme predictions in related bacteria and for engineering enzymes for biotechnology or biomedical applications. [ABSTRACT FROM AUTHOR]

This letter discusses two cases of classic galactosemia in infants that presented with pseudohyperglycemia due to interference from a glucometer. The cases highlight the limitations of point-of-care glucometers for diagnosing hyperglycemia and emphasize the need for confirmation using an alternative method, such as a hexokinase assay in the central laboratory. Galactosemia should be considered in the differential diagnosis for infants with pseudohyperglycemia. The letter also mentions the risks of iatrogenic insulin-induced hypoglycemia and the FDA's notification about potentially fatal errors with GDH-PQQ glucose monitoring technology. [Extracted from the article]

The marine sponge Hymeniacidon perlevis is a globally distributed and invasive species with extensive filter-feeding characteristics. The symbiotic relationship fostered between the sea sponge and the inhabiting microorganism is key in the production of metabolic enzymes which is the focus of this study. Sponge bacterial symbionts were grown on starch agar for 48hrs. Colourimetric analyses of amylase were conducted at 540nm using a spectrophotometric plate reader. Using an X-Bridge column (3.5μM, 4.6x150mm), 80/20 acetonitrile/water in 0.1% ammonium were the conditions used for the liquid chromatography-mass spectrometry (LC-MS) analyses. Seven reducing sugars were used to optimise LC-MS to determine the presence of the crude enzyme formed. Not all the bacterial symbionts isolated from H perlevis produced alpha and beta amylases to break down starch. From the statistical mean of crude enzyme concentrations from the hydrolysis of starch by amylase, isolate seven had the highest optical density (OD) at 0.43475 while isolate twelve had the lowest OD at 0.141417. From the LC-MS analysis, out of the seven sugars, Glucose and maltose constituted > 65% of the reducing sugars formed from the hydrolysis of starch by the amylases. Isolates 3,6 and 7 produced 6.906 mg/l, 12.309 mg/l, and 5.909 mg/l of glucose, while isolates 3,4,5,6 and 7 produced 203.391 mg/l, 176.238 mg/l, 139.938 mg/l, 39.030 mg/l, and 18.809 mg/l of maltose, respectively. Isolate two had the highest amount of maltose at a concentration of 267.237 mg/l while isolate four had the highest amount of glucose concentration of 53.084 mg/l. Enzymes from marine sponge bacteria offer greater potential for a green and sustainable production process. Amylase extraction from bacterial symbionts in H perlevis is sustainable and should be supported. They can serve as reliable sources of revenue for enzyme industries, and applications in food industries and biotechnological processes. [ABSTRACT FROM AUTHOR]