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Oxidation destroys the flavor and freshness of the beer, and yeast with improved antioxidant activity would benefit the flavor stability of the beer. The aim of this work was to study the antioxidant networks of lager yeast for brewing industry. Through transcriptome and metabolome analysis, we were able to map changes in anti-oxidant pathways and transcription of genes in lager yeast. Results suggested that metabolic energy variation in yeast might be the main reason for the improved anti-oxidant capacity. Lower metabolic rate kept the yeast at relatively lower respiratory status at the end of fermentation, thereby prolonging the lifespan and potentially supporting its usage in serial fermentations. Up-regulation of mannose synthesis and hexose transport strengthened the cell structure, which may have contributed to the anti-oxidant capacity of yeast. A deeper understanding of the globally regulation pattern of anti-oxidation in lager yeast is expected to support the design of strain development strategies for improved anti-oxidant capacity of yeast for fermentation industry.

Lactobacillus species are the main spoilage bacteria in beer production and storage. The prerequisite for Lactobacillus species to grow in beer is to be able to resist the hop bitter compound iso-α...

Ovarian cancer (OC) is the most frequent cause of death among patients with gynecologic malignancies. In recent years, the development of cisplatin (DDP) resistance has become an important reason for the poor prognosis of OC patients. Therefore, it is vital to explore the mechanism of DDP resistance in OC. In this study, microRNA-1246 (miR-1246) expression in OC and DDP-resistant OC cells was determined by RT-qPCR, and chemosensitivity to DDP was assessed by the CCK-8 assay. A dual-luciferase reporter assay was performed to confirm the interaction between miR-1246 and zinc finger 23 (ZNF23), while changes in ZNF23 expression were monitored by RT-qPCR, immunofluorescence, and western blot assays. Moreover, cell proliferation, cycle phase, and apoptosis were determined by EdU staining, flow cytometry, TUNEL staining, and Hoechst staining. Our data showed that miR-1246 was highly expressed in DDP-resistant OVCAR-3 and TOV-112D cells. Functionally, overexpression of miR-1246 markedly enhanced DDP resistance and cell proliferation, and suppressed cell cycle arrest and apoptosis of OC cells. Inhibition of miR-1246 expression significantly attenuated DDP resistance and cell proliferation, and increased cell cycle arrest and apoptosis in DDP-resistant OC cells. Furthermore, ZNF23 was identified as a target gene of miR-1246, and ZNF23 protein expression was notably downregulated in DDP-resistant OC cells. Moreover, overexpression of miR-1246 significantly downregulated the ZNF23 levels in OVCAR-3 and TOV-112D cells, and inhibition of miR-1246 upregulated the ZNF23 levels in the DDP-resistant OVCAR-3 and TOV-112D cells. In conclusion, miR-1246 might be a novel regulator of DDP-resistant OC that functions by regulating ZNF23 expression in DDP-resistant cells, as well as cell proliferation, cell cycle progression, and apoptosis.

Biogenic amines (BAs) are a threat to the safety of broad bean paste, and biosynthetic mechanism of BA and its regulation are unknown. This study aimed to assess microbial BA synthesis in Chinese traditional broad bean paste and determine favorable fermentation conditions for BA regulation. The BAs content in 27 pastes was within the safe range. 64 strains with potential decarboxylation were screened in Luria–Bertani Glycerol medium and identified as Bacillus spp. Although Bacillus amyloliquefaciens produced highest levels of BAs (70.14 ± 2.69 mg/L) in LBAA, Bacillus subtilis produced 6% more BAs than B. amyloliquefaciens. Meanwhile, temperature was the most remarkable factor affecting BAs production by B. amyloliquefaciens 1–13. Furthermore, the fermented broad bean paste model revealed that BA content increased by 61.2 mg/kg every 10 days at 45 °C, which was approximately threefold of that at 25 °C. An ARIMA prediction model of BAs content was constructed, and the total BAs content of 40 mg/100 g was set as the critical value. This study not only contributed to understanding the BAs formation mechanism, but also provided potential measures to control the BAs in fermented soybean products. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13197-020-04781-z) contains supplementary material, which is available to authorized users.

As one of the most common cancers in female, ovarian cancer (OC) has become a serious public burden now. Mounting researches have indicated long noncoding RNAs (lncRNAs) can affect many biological processes including cancer development. LncRNA LBX2-AS1 was identified to be an oncogene in some cancers, but the role of LBX2-AS1 in OC remains to be elucidated. Bioinformatics analysis and experiments including ChIP, RT-qPCR, RIP, luciferase reporter, western blot and CCK-8 were performed to explore the role of LBX2-AS1 in OC. LBX2-AS1 expression was markedly increased in OC tissues and cell lines. Functionally, LBX2-AS1 silencing inhibited cell proliferation, migration and stemness but facilitated cell apoptosis in OC. Moreover, depletion of LBX2-AS1 suppressed tumor growth of OC in vivo. Mechanically, LBX2-AS1 was activated by transcriptional factor ELK1. ELK1 enhanced the expression of LBX2-AS1 in OC cells. In addition, miR-4784 was confirmed to be sponged by LBX2-AS1. There was a negative expression correlation between LBX2-AS1 and miR-4784 in OC tissues. Subsequently, KDM5C was identified to be a direct target of miR-4784 in OC cells. KDM5C was negatively regulated by miR-4784 and positively regulated by LBX2-AS1 in terms of expression level. Upregulation of KDM5C reversed the inhibitory effect of LBX2-AS1 depletion on the progression of OC. This study proved that ELK1 activated-LBX2-AS1 aggravated the progression of OC by targeting the miR-4784/KDM5C axis, suggesting that LBX2-AS2 may be a promising diagnostic biomarker of OC.

This study aimed to address the roles of divalent-cation transporter genes of Lactobacillus casei 2-9-5 in resisting hop bitter compound iso-α-acid. To determine the underlying mechanisms for Lacto...

Lager yeast is the most popular yeast strain used for beer production in China. The flocculation of yeast plays an important role in cell separation at the end of fermentation. Therefore, appropriately enhancing the flocculation capability of the lager yeast without affecting its fermentation performance would be desirable for beer industry. Our previous study showed that the defect of gene RIM21 might contribute to the enhanced flocculation capability of a lager yeast G03. To further investigate the role of the RIM21 gene in flocculation of strain G03, this study constructed a RIM21-deleted mutant strain G03-RIM21Δ through homologous recombination. Deletion of RIM21 improved the flocculation capability of strain G03 during wort fermentation at 11 °C without changing its fermentation performance significantly. The expression of FLO5, Lg-FLO1 and some other genes involved in cell wall integrity pathway were up-regulated in strain G03-RIM21Δ. In addition, the disruption of RIM21 enhanced resistance of yeast cells to cell wall inhibitors. These results provide a basis for elucidating the flocculation mechanism of lager yeast under low-temperature fermentation conditions.

Objective To evaluate real-world use and outcomes of apatinib treatment in platinum-resistant recurrent epithelial ovarian cancer. Methods This is an observational study. Patients with platinum-resistant recurrent epithelial ovarian cancer initiating apatinib treatment from January 2016 to December 2018 were included. The primary end point was progression-free survival. Other end points included overall survival, objective response rate, disease control rate, and toxicity. Results A total of 28 platinum-resistant epithelial ovarian cancer patients were enrolled in this study. Thirteen cases received apatinib as maintenance therapy following chemotherapy with a median progression-free survival of 6.0 months and a medium overall survival of 11.0 months. Four patients received apatinib as palliative following chemotherapy with 2 cases in progressive disease and 2 cases in stable disease. Eleven cases received apatinib alone as salvage therapy with a disease control rate of 81.8% and a median progression-free survival of 3.0 months. The most common adverse effects were hand-foot syndrome (53.57%), secondary hypertension (46.43%) and fatigue (14.29%). Five patients discontinued treatment due to grade 3 toxicities and 4 patients required dose reduction because of adverse effects. Conclusion Apatinib produced moderate improvements in progression-free survival in patients with platinum-resistant epithelial ovarian cancer both as maintenance therapy following chemotherapy and as single-agent salvage therapy. Our study suggests that apatinib may be effective for women with platinum-resistant recurrent epithelial ovarian cancer.

This study aimed to elaborate the role of sunlight on chemosensory characteristics of broad bean paste (BBP). Two groups of BBP fermentation with sunlight treated (ST) or darkness treated (DT) were prepared and the volatile compounds were revealed and compared via GC-MS and GC-IMS and multivariate statistical analysis. A total of 125 compounds were identified and 24 markers with potential contribution to the BBP flavor characteristics were screened. Sunlight exposure was proposed to favor the accumulation of volatile compounds providing fruity, floral, buttery and roasted aroma during fermentation while volatile flavor compounds with unfavorable aroma were enriched in the darkness-treated group. Through KEGG-enrichment analysis, phenylalanine metabolism was found to play critical roles in regulating volatile flavor compound formation and the predicted synthesis network for 24 marker volatile flavor compounds was constructed. This study may unravel the scientific meaning of the role of sunlight on BBP chemosensory quality.

Flavor stability is a significant concern to brewers as the staling compounds impart unpleasant flavor to beer. Thus, yeasts with antistaling ability have been engineered to produce beer with improved flavor stability. Here, we proposed that increasing the NADH availability of yeast could improve the flavor stability of beer. By engineering endogenous pathways, we obtained an array of yeast strains with a higher reducing activity. Then, we carried out beer fermentation with these strains and found that the antistaling capacities of the beer samples were improved. For a better understanding of the underlying mechanism, we compared the flavor profiles of these strains. The production of staling components was significantly decreased, whereas the content of antistaling components, such as SO

Purpose Ovarian cancer is the most lethal cancer among all gynaecological malignancies. The combination theraputics of cisplatin and taxol is widely used in clinicals for ovarian cancer treatment. However, long-term use of cisplatin and taxol induces strong tolerance and hepatotoxicity. Since silibinin is a commonly used anti-hepatotoxic drug in Europe and Asia, the aim of this study was to determine whether silibinin could restore the sensitivity of combination use of cisplatin and taxol in drug-resistant human ovarian cancer cells and reduce drug-induced hepatotoxicity. Patients and methods Normal hepatocyte LO2 cells and A2780/DDP cells were treated with silibinin, cisplatin, taxol, cisplatin and taxol plus silibinin for 48 h. Cell viability was determined by MTT and long-term proliferation assay, while apoptosis and cell cycle progression were assessed by flow cytometric analysis. DNA damage was evluated by immunofluorescence assays. The metastatic activity of A2780/DDP was determined by cell adhesion assay. Results The addition of silibinin on cisplatin and/or toxal could sensitize the antitumor activity of cisplatin and toxal on A2780/DDP cells, supress cell-matrix adhesion of A2780/DDP, inhibit the cell proliferation, result in A2780/DDP cells apoptosis. In addition, silibinin could effectively reduce cisplatin and/or toxal-induced hepatotoxicity by protecting DNA from damage and restoring the potential of cell proliferation in cisplatin and/or toxal-treated LO2 cells. Conclusion Our results suggest that silibinin could restore the sensitivity of cisplatin and taxol in drug-resistant human ovarian cancer cells and reduce durg-induced hepatotoxicity in cell level.

Precise determination of yeast physiology is crucial in both laboratories and breweries. Our aim was to identify a sensitive indicator that could demonstrate subtle physiological differences in bre...

To improve the aroma profile of beer by using metabolic engineering to increase the availability of cytosolic NADH in lager yeast. To alter NADH levels in lager yeast, the native FDH1 (YOR388C) encoding NAD+-dependent formate dehydrogenase was overexpressed in the yeast strain M14, yielding strain M-FDH1. This led to a simultaneous increase of NADH availability and NADH/NAD+ ratio in the M-FDH1 strain during fermentation. At the end of the main fermentation period, ethanol production by strain M-FDH1 was decreased by 13.2%, while glycerol production was enhanced by 129.4%, compared to the parental strain respectively. The production of esters and fusel alcohols by strains M14 and M-FDH1 was similar. By contrast, strain M-FDH1 generally produced less organic acids and off-flavor components than strain M14, improving the beer aroma. Increased NADH availability led to rerouting of the carbon flux toward NADH-consuming pathways and accelerated the NADH-dependent reducing reactions in yeast, greatly impacting the formation of aroma compounds and improving the beer aroma.

Serial repitching of beer brewer’s yeast plays an important role in the beer industry as an ineluctable economic factor. In this study, the viability and vitality changes, as well as strains’ anti-autolytic abilities during serial beer fermentation with typical ale and lager yeast strains, were investigated. While measuring the survival rate of yeast cells is not sufficient for evaluating and predicting the yeast fermentation capacity, physiological status determination of brewer’s yeast reflects the vitality and quality of yeast in serial beer fermentations. Accumulation of reactive oxygen species in yeast causes cell damage, leading to a decline in cell vitality and viability. Aged yeast cells, after several repitchings, can result in leakage of intracellular compounds into the fermented liquor. The lager yeast (Pilsner) examined, which harbors the partial genome from S. eubayanus, showed better robustness and higher activity than the ale yeast strain M79 examined, during serial fermentation. A holistic approach, including more indicators, should be applied in evaluating the fermentation performance of brewer’s yeast.

Acetaldehyde is synthesized by yeast during the main fermentation period of beer production, which causes an unpleasant off-flavor. Therefore, there has been extensive effort toward reducing acetaldehyde to obtain a beer product with better flavor and anti-staling ability. In this study, we discovered that acetaldehyde production in beer brewing is closely related with the intracellular NADH equivalent regulated by the citric acid cycle. However, there was no significant relationship between acetaldehyde production and amino acid metabolism. A reverse engineering strategy to increase the intracellular NADH/NAD+ ratio reduced the final acetaldehyde production level, and vice versa. This work offers new insight into acetaldehyde metabolism and further provides efficient strategies for reducing acetaldehyde production by the regulating the intracellular NADH/NAD+ ratio through cofactor engineering.

Ethyl-acetate is important for the flavor and aroma of the alcoholic beverages, therefore, there have been extensive efforts toward increasing its production by engineering yeast strains. In this study, we reported a new approach to breed non-genetic modified producing yeast strain with higher ethyl-acetate production for beer brewing. First, we demonstrated the positive effect of higher acetic acid concentration on inducing the expression of ACS. Then, we applied adaptive laboratory evolution method to evolve strain with higher expression level of ACS. As a result, we obtained several strains with increased ACS expression level as well as ethyl-acetate production. In 3 L scale fermentation, the optimal strain EA60 synthesized more ethyl-acetate than M14 at the same time point. At the end of fermentation, the ethyl-acetate production in EA60 was 21.4% higher than M14, while the other flavor components except for acetic acid were changed in a moderate degree, indicating this strain had a bright prospect in industrial application. Moreover, this study also indicated that ACS1 played a more important role in increasing the acetic acid tolerance of yeast, while ACS2 contributed to the synthesis of cytosol acetyl-CoA, thereby facilitating the production of ethyl-acetate during fermentation.

High stability at acidic environment is required for 1,3-1,4-β-glucanase to function in biofuel, brewing and animal feed industries. In this study, a mesophilic β-glucanase from Bacillus terquilensis CGX 5-1 was rationally engineered through sequence alignment and surface charge engineering to improve its acidic resistance ability. Nineteen singly-site variants were constructed and Q1E, I133L and V134A variants showed better acidic stability without the compromise of catalytic property and thermostability. Furthermore, four multi-site variants were constructed and one double-site variant Q1E/I133L with better stability at acidic environment and higher catalytic property was obtained. The fluorescence spectroscopy and structural analysis showed that more surface negative charge, decreased exposure degree of residue No.1, shifted side chain direction of residue No.133 and the lower total and folding free energy might be the reason for the improvement of acidic stability of Q1E/I133L variant. The obtained Q1E/I133L variant has potential applications in industries.

The fermentation performance of yeast is the key of beer production. High gravity brewing is a commonly used technique in industrial lager beer production and it is environmentally friendly. Therefore, there has been extensive effort toward improving high gravity brewing. In this study, through transcriptomic and metabolomic analysis of two homologous lager yeasts, genes that relate to stress tolerance in high gravity brewing were screened. The results showed EMP pathway and multiple amino acid metabolism pathway were the most enriched pathways, and pyruvate might be the core metabolite. Overexpression and knockdown strains were constructed to verify the genes' functions. The overexpression of MAN2, PCL1 and PFK26 genes were beneficial to fermentation without significantly changes in flavor profiles. The relative intracellular ATP levels can help us understand the change of metabolic flux such as enhancement of sugar consumption. This work is helpful to reveal the stress tolerance mechanism of high gravity brewing and breed yeast strains with improved performance.

Yeast flocculation plays an essential role in industrial application. Appropriate flocculation of yeast cells at the end of fermentation benefits the cell separation in production, which is an important characteristic of lager yeast for beer production. Due to the complex fermentation environment and diverse genetic background of yeast strains, it is difficult to explain the flocculation mechanism and find key genes that affect yeast flocculation during beer brewing. By analyzing the genomic mutation of two natural mutant yeasts with stronger flocculation ability compared to the parental strain, it was found that the mutated genes common in both mutants were enriched in protein processing in endoplasmic reticulum, membrane lipid metabolism and other pathways or biological processes involved in stress responses. Further functional verification of genes revealed that regulation of RIM101 and VPS36 played a role in lager yeast flocculation under the brewing condition. This work provided new clues for improving yeast flocculation in beer brewing.

Background: Despite advancements in drug therapy, many patients with adenomyosis fail to achieve treatment goals. Therefore, new therapies need to be developed. Our aim was to evaluate the efficacy, safety and tolerability of mifepristone versus placebo in the treatment of painful adenomyosis. Methods: We conducted a randomized, multicenter, double-blind and placebo-controlled trial in 10 hospitals in China. Eligible patients were randomly assigned in a 1:1 ratio to receive mifepristone or placebo orally once a day for 12 weeks. The randomization sequence was generated by the block-rank package of R software with a block size of four. Randomization was performed centrally with an interactive web response system. Regular trial visits were performed every 4 weeks and lasted to 4 weeks after treatment. Patients, investigators, clinicians and statisticians were masked to the treatment allocation throughout the study. The primary outcome was the change in adenomyosis-associated dysmenorrhea intensity evaluated by visual analog scale (VAS) in intention-to-treat population. Secondary outcomes were the changes in menstrual blood loss, the levels of hemoglobin and CA125, platelet count and uterine volume after 12-week treatment. Safety analyses were done in patients with at least one dose of the product intervention. This trial is registered with ClinicalTrials.gov, NCT03520439. Findings: Between May 2018 and April 2019, 134 adenomyosis patients with dysmenorrhea were randomly assigned. Eight patients without any valid data at baseline were excluded and 126 patients were included in efficacy analysis (61 to mifepristone and 65 to placebo). The mean changes in VAS score were −6.63 (standard deviation [SD] 1.92) in mifepristone group and −0.95 (SD 1.75) in placebo group (p < 0.0001). The total effective and complete remission rates for dysmenorrhea in mifepristone group were significantly higher than those in placebo group ( 91.80% vs. 23.08% and 88.52% vs. 6.15%, respectively ). All the secondary outcomes showed significant improvements after mifepristone treatment for menstrual blood loss, the levels of hemoglobin and CA125, platelet count and uterine volume (P < 0.0001 for all). Safety analysis revealed no significant difference between two groups, and no serious adverse events were reported. Interpretation: Treatment with mifepristone resulted in significantly lower pain scores for adenomyosis patients with dysmenorrhea . Secondary outcomes were all met with a significant improvement with no new safety concerns and well-toleration. These data support that mifepristone could serve as a new treatment option for adenomyosis patients to relieve the symptoms with safety. Trial Registration: This trial is registered with ClinicalTrials.gov, NCT03520439. Funding/Support: This study was supported by National Key R&D Program of China (Grant number: 2017YFC1001202), National Natural Science Foundation of China (Grant numbers: 81974225, 81671429 and 81802591), Research Fund for Lin He’s Academician Workstation of New Medicine and Clinical Translation (Grant number: 19331203) and Jiaxing Municipal Public Welfare Research Project (Grant number: 2021AY30004). Declaration of Interest: None to declare. Ethical Approval: Ethical approval was obtained by the institutional review board or ethics committee at each investigational site before the initiation of the trial.

Background: To describe the characteristic of plasma lipid prolife among cervical cancer and further evaluate the prognostic significance of lipid levels in cervical cancer. Methods: We retrospectively reviewed 1,713 patients with cervical cancer in our hospital. Preoperative plasma lipid profile, including cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL), and low-density lipoprotein cholesterol (LDL), of 1,713 cervical cancer patients were compared with 10,397 healthy women. Then, we evaluated the impact of lipids on overall survival (OS) and recurrence-free survival (RFS) in cervical cancer using univariate and multivariate Cox model. Results: While plasma TC, TG and LDL were significantly higher, HDL was lower in patients with cervical cancer as compared with healthy women. TG was identified as independent predictors for RFS, and OS among patients with cervical cancer. Further stratified by age, patients with higher TG showed a significantly worse RFS and OS compared with those with lower TG among patients ≥50 years old, but not among Conclusion: Cervical cancer was associated with a disordered lipid profile. Hypertriglyceridemia was an independent poor prognostic indicator for cervical cancer, especially for elder patients. Strengthening lipid management may be beneficial for improving postoperative OS and RFS in patients with cervical cancer.

Bean-based fermentation foods are usually ripened in open environment, which would lead to inconsistencies in flavor and quality between batches. The physicochemical metabolism and microbial community of seasonal broad bean paste (BBP) were compared to distinguish discriminant metabolites and unique taxa, as well as their specific reasons for different flavor and quality in this study. Here, we found that environmental variables led to the seasonal distribution of microbiota, and differential microorganisms further contributed to the inconsistency of flavor quality, in which Lactobacillales was responsible for the higher titratable acid and amino acid nitrogen concentration in winter pei, while Saccharomycetales benefited the formation of volatile flavor substances in autumn pei. Additionally, we compared the effect of different combinations of Lactobacillales with Zygosaccharomyces rouxii on the quality of BBP, and found that W. confusa was more suitable for BBP fermentation rather than T. halophilus in terms of sensory characteristics and physicochemical metabolites.

This study aimed to elaborate the roles of salt concentration on doubanjiang (broad bean paste) fermentation. Three sets of doubanjiang samples which had lower salt concentration than commercial doubanjiang were prepared and the physicochemical parameters, biogenic amines, flavor, microbial dynamics were analyzed during fermentation. The salt reduction showed significant effect on the dynamics of bacteria and fungi, thus leading to doubanjiang samples with different properties. Salt reduction during fermentation relieved the osmotic pressure towards microbes, which favored the accumulation of amino acid nitrogen, amino acids, and volatile flavor compounds. However, higher concentrations of total acids and biogenic amines and the existence of conditional pathogens, such as Klebsiella, Cronobacter and Acinetobacter genera, were observed in salt reduced doubanjiang samples, which was undesirable for doubanjiang quality. This study would deep our understanding of the roles of salt on doubanjiang fermentation.

Brewer’s yeast has been widely used in the food industry, and the autolysates thereof are increasingly being studied for their valuable nutritional compositions. Yeast autolysis is most affected by medium composition and temperature. In this study, a thermosensitive autolytic brewer’s yeast P-510 was obtained with atmospheric and room temperature plasma mutagenesis plus 5-bromo-chloro-3-indolyl phosphate screening. The mutant rapidly autolyzed at 37 °C and the autolysates contained more active components and showed higher antioxidant activities compared with that of the parental strain, which indicated that the mutant’s autolysates can potentially be used as functional food and nutritional ingredients. Transcriptomic analysis of the mutant and parental strains at 28 and 37 °C suggested that thermosensitive autolysis of P-510 was probably caused by mitochondrial disfunction, glycogen metabolic flux of glycolysis and pentose phosphate pathway disorder, as well as hexose transport inhibition. The results revealed the important role of mitochondrial metabolism and glycogen utilization regulation in heat stress response of yeast.

White spots are commonly found in bean-based fermented food, which will significantly lower the product quality. This study aimed to analyze the composition of white spots and further reveal the source and influencing factors of white spots in bean-based fermented food using soybean paste as study model. The results showed that white spots were mainly composed of 40.96% free tyrosine and 37.94% tyrosine in combination form. During soybean paste fermentation, tyrosine was found to be produced by the actions of proteolytic enzymes secreted by Aspergillus oryzae 3.042 instead of the microbial metabolism and the excessive accumulation of tyrosine in soybean paste led to the formation of white spots. Among all influencing factors, high temperature treatment favored the formation of white spots. The existence of soy peptone and phenylalanine would postpone the precipitation of tyrosine while promoting the aggregation of the tyrosine precipitation. Field emission scanning electron microscope analysis showed that tyrosine would accumulate around the soybean protein particles and treatment at 120 °C would disrupt the structure of tyrosine-protein complex. Based on the above results, we proposed that treatment of soybean paste at temperature lower than 80 °C was the current practically applicable method to control the formation of white spots in soybean paste. PRACTICAL APPLICATION: This study developed a new idea to understand the composition and formation of white spots in soybean paste, which would provide guidance for prevention and control of white spots during the production of soybean paste for manufacturers and researchers.

The role of proteins, especially protein Z, on beer foam stability was studied. The results showed that both positive and negative proteins were enriched in beer foam. Contrary to previous results, proteins with lower surface hydrophobicity were preferred for beer foam stability. Among all proteins, protein Z was the most prevalent protein in beer foam and could be accurately measured using an enzyme linked immunosorbent assay. The quantity of protein Z was found to be positively correlated with beer foam stability. The addition of purified protein Z from barley (Hordeum vulgare) malt into the finished beer could enhance the beer foam stability. Therefore, proteins with lower surface hydrophobicity might be beneficial for beer foam stability, especially protein Z.

Saccharomyces cerevisiae strains with favorable characteristics are preferred for application in industries. However, the current ability to reprogram a yeast cell on the genome scale is limited due to the complexity of yeast ploids. In this study, a method named genome replication engineering-assisted continuous evolution (GREACE) was proved efficient in engineering S. cerevisiae with different ploids. Through iterative cycles of culture coupled with selection, GREACE could continuously improve the target traits of yeast by accumulating beneficial genetic modification in genome. The application of GREACE greatly improved the tolerance of yeast against acetic acid compared with their parent strain. This method could also be employed to improve yeast aroma profile and the phenotype could be stably inherited to the offspring. Therefore, GREACE method was efficient in S. cerevisiae engineering and it could be further used to evolve yeast with other specific characteristics.

Autophagy is a survival process that maintains homeostasis in all eukaryotic cells. Recent studies show an abnormal autophagic activity in endometriosis, but the role of autophagy is controversial. Homeobox A10 (HOXA10) is a transcription factor necessary for embryonic and adult uterine development, and studies indicate that its expression decreases in endometriosis. Homeobox A10 may negatively regulate autophagy in endometriosis. To test this hypothesis, we measured the expression levels of autophagic biomarkers (beclin-1 and LC3-II) and HOXA10 proteins by Western blotting and messenger RNA (mRNA) by quantitative real-time polymerase chain reaction. Furthermore, we evaluated the serum cancer antigen 125 (CA125) levels by immunoassay. Most tested autophagic biomarker proteins and mRNAs were upregulated, whereas HOXA10 protein and mRNA were decreased in ovarian endometriomas compared with eutopic endometria of women with endometriosis and normal endometria. Compared with normal endometrium, only protein expression levels of autophagic biomarkers were increased in the eutopic endometrium of women with endometriosis. Moreover, HOXA10 was found to have a significant negative correlation with autophagy ( P < .01). Serum CA125 was at a high level in endometriosis and increased with elevated revised American Fertility Society staging (I-IV). There was a significant positive correlation between serum CA125 level and LC3-II protein level and/or LC3-II/LC3-I ratio ( P < .01) and a significant negative correlation between serum CA125 level and HOXA10 gene level ( P < .01). In conclusion, our studies support that the deficiency of HOXA10 may induce autophagy in endometriosis, and the relationship among CA125, autophagy, and HOXA10 in endometriosis requires additional research.

Acetaldehyde is an important flavor component in beer which is possibly carcinogenic to humans. Owing to the limitations of present detection methods, only free-state acetaldehyde in beers has been focused on, while acetal in beers has hardly been reported so far.; Results: A sensitive headspace gas chromatography method was developed for the determination of diethylacetal and acetaldehyde in beer. The column DB-23 was chosen with a total run time of 22.5 min. The optimal addition amount of NaCl, equilibrium temperature and equilibrium time were 2.0 g, 70 °C and 30 min respectively. For both diethylacetal and acetaldehyde analyses, the limit of detection was 0.005 mg L-1 with relative standard deviation < 5.5%. The recoveries of acetaldehyde and diethylacetal were 95-110 and 95-115% respectively. The diethylacetal and acetaldehyde average contents in 24 beer products were 11.83 and 4.36 mg L-1 respectively. The Pearson correlation coefficient between diethylacetal and acetaldehyde was the highest (0.963). Both diethylacetal and acetaldehyde contents increased to a peak value after fermentation for 3 days and then decreased to a lower value. During both normal and forced aging storage, the diethylacetal content decreased and the acetaldehyde content increased gradually over time. When beers were forced aged for 4 days, the increased ratio of acetaldehyde could be above 40.00%.; Conclusion: The newly established method can be used to assess acetaldehyde level and flavor quality in beer more scientifically. © 2018 Society of Chemical Industry.; © 2018 Society of Chemical Industry.

β-Amylase is of important biotechnological aid in maltose syrup production. In this study, the extraction condition of β-amylase from brewer's malt and the optimal dosage of β-amylase in maltose syrup production were optimized using response surface methodology and uniform design method. The optimal extraction condition of β-amylase from brewer's malt was composed of 1:17 (g/v) material/liquid ratio, 44°C extraction temperature, pH 6.4 buffer pH, 2.3 H extraction time, and 1.64 g L-1 NaSO3 dosage with a predicted β-amylase activity of 1,290.99 U g-1 , which was close to the experimental β-amylase activity of 1,230.22 U g-1 . The optimal dosages of β-amylase used in maltose syrup production were 455.67 U g-1 starch and its application in maltose syrup production led to a 68.37% maltose content in maltose syrup, which was 11.2% and 28.9% higher than those using β-amylases from soybean and microbe (P

This study aimed to isolate and identify the spoilage microbes which could produce gas in fermented broad bean paste and to provide an effective method to restrain their growth in media. The gas-producing spoilage microbe was isolated in anaerobic condition and identified as Bacillus licheniformis FHYM 8. B. licheniformis FHYM 8 was resistant to high temperature, low pH and high salinity condition since it could survive after treatment at 100 °C for 10 min and when the pH value was as low as pH3.8. The inhibitory salinity for B. licheniformis FHYM 8 was 15% in aerobic condition while 11% of salinity could completely inhibit its growth in anaerobic condition. Interestingly, B. licheniformis FHYM 8 showed the ability to produce gas when the broad bean paste was sterilized at 66 °C while no gas was produced when B. licheniformis FHYM 8 was inoculated in broad bean paste after the heat treatment process, indicating that heat treatment was the necessary step for B. licheniformis FHYM 8 to produce CO 2 in broad bean paste. The inhibition analysis showed that nisin with a minimal inhibitory concentration of 0.02 mg mL −1 was efficient in inhibiting the growth of B. licheniformis FHYM 8 in MRS media when the salinity was 8%. This will provide guidance to broad bean paste manufacturers to further adopt efficient strategies to inhibit the growth of gas-producing microbe in broad bean paste.

1,3-1,4-β-glucanase was an important biotechnological aid in the brewing industry. In a previous research, a Bacillus BglTO mutant (BglTO) with high tolerance towards high temperature and low-pH conditions was constructed and expressed in Escherichia coli. However, E. coli was not a suitable host for enzyme production in food industry. Therefore, the present work aimed to achieve the high-level expression of BglTO in Bacillus subtilis WB600 and to test its effect in Congress mashing. The β-glucanase mutant was successfully expressed in B. subtilis WB600 and favorable plasmid segregation and structural stability were observed. The maximal extracellular activity of β-glucanase in recombinant B. subtilis WB600 reached 4840.4 U mL−1 after cultivation condition optimization, which was 1.94-fold higher than that before optimization. The fermentation capacity of recombinant B. subtilis reached 242.02 U mL−1 h−1, which was the highest among all reported β-glucanases. The addition of BglTO in Congress mashing significantly reduced the filtration time and viscosity of mash by 29.7% and 12.3%, respectively, which was superior to two commercial enzymes. These favorable properties indicated that B. subtilis WB600 was a suitable host for production of BglTO, which was promising for application in the brewing industry.

The aim of this study was to discuss the functions of FKS family genes which encode β-1, 3-glucan synthase regarding the viability and autolysis of yeast strain. Loss of FKS1 gene severely influences the viability and anti-autolytic ability of yeast. Mutation of FKS1 and FKS2 genes led to cell reconstruction, resulting in a sharp shrinkage of cell volume and decreased stress resistance, viability, and anti-autolytic ability. Deletion of FKS3 gene did not clearly influence the synthesis of β-1, 3-glucan of yeast but increased the strain's stress resistance, viability, and anti-autolytic ability. It is suggested that FKS3 would be the potential target for improving the stress resistance of yeast. The results revealed the relationship among FKS family genes and demonstrated their functions on yeast cell wall construction and anti-autolytic ability.

This study aimed to elaborate the effect of N-glycosylation and glycation of protein Z4 from barley malt and recombinant Pichia pastoris on beer foam stability. The malt protein Z4 and recombinant protein Z4 showed similar N-glycosylation patterns while recombinant protein Z4 was glycosylated at a higher degree. In the simulated mashing and boiling, malt protein Z4 and deglycosylated malt protein Z4 preferred to glycate with glucose and maltose while recombinant protein Z4 and deglycosylated recombinant protein Z4 showed preference towards fructose. The addition of protein Z4 and protein Z4-saccharide complexes in finished beer showed that the addition of glycosylated protein Z4 only slightly enhanced the beer foam stability while the addition of glycated protein Z4 and protein Z4 with both glycation and glycosylation could significantly increase the beer foam stability. Therefore, glycation instead of N-glycosylation of protein Z4 played important roles in maintaining beer foam stability.

Although the microbial diversity and structure in bean-based fermented foods have been widely studied, systematic studies on functional microbiota and mechanism of community forms in multi-microbial fermentation systems were still lacking. In this work, the metabolic pathway and functional potential of microbial community in broad bean paste (BBP) were investigated by metagenomics approach, and Staphylococcus, Bacillus, Weissella, Aspergillus and Zygosaccharomyces were found to be the potential predominant populations responsible for substrate alteration and flavor biosynthesis. Among them, Staphylococcus was the most abundant and widespread functional microbe, and closely related Staphylococcus species were diverse and ubiquitously distributed, with the opportunistic pathogen S. gallinarum being the most abundant Staphylococcus specie isolated from BBP. To explain the dominance status of S. gallinarum and species distributions of Staphylococcus genus, we tested the effects of abiotic and biotic factors on three Staphylococcus species using a tractable BBP model, demonstrating that adaptation to environmental conditions (environmental parameters and other functional microbes) led to the dominant position and species coexistence of Staphylococcus, and congeneric competition among Staphylococcus species further shaped ecological distributions of closely related Staphylococcus species. In general, this work revealed the metabolic potential of microbial community and distribution mechanism of Staphylococcus species during BBP fermentation, which could help traditional factories to more precisely control the safety and quality of bean-based fermented foods.

‘Bask in daytime and dewed at night’ technique is traditionally used in doubanjiang fermentation, however, the underlying mechanism behind this technique is still unrevealed. This study aimed to elaborate the roles of three key indexes in this technique, that are air, sunlight, and mash turnover, on doubanjiang quality through the design of doubanjiang fermentation. The results showed that the exposure to air or sunlight was beneficial for doubanjiang to yield reddish-brown luster color, less acidity and balanced flavor. On the contrary, blocking of air and sunlight could accumulate higher acidity (4.75 ± 0.01 g/100gdw), amino nitrogen (2.08 ± 0.07 g/100gdw), alcohols (36.9 g/kg) and esters (26.9 g/kg). Regular mash turnover was mainly involved in making different mash layers exposure to air or sunlight equally through homogenizing mash layers. Different conditions also have been suggested to affect the microbiota dynamics by amplicon sequencing and RT-qPCR. Based on RDA analysis, Pediococcus, Meyerozyma, Bacillus and Zygosaccharomyces were predicted to be related to the changes of physicochemical properties or volatile compounds in different conditions, and their function was validated by simulated fermentation. This study would unravel the underlying scientific meaning of ‘bask in daytime and dewed at night’ technique in doubanjiang fermentation.

Aim A previous study reported that LINC00261 is significantly downregulated in human ectopic endometrial tissues. The present study aimed to explore whether LINC00261 is functional in endometriosis cell proliferation, apoptosis and migration. Methods By transfecting human endometriosis cell line CRL-7566 with plasmids containing LINC00261, we successfully established the cell CRL-7566/LINC00261 with a high LINC00261 expression level. Cell-counting kit-8 and colony formation assays were conducted to evaluate the effect of LINC00261 on cell proliferation, and flow cytometry analysis and transwell migration assay were conducted to evaluate its effect on cell apoptosis and cell migration, respectively. Results Cell-counting kit-8 and colony formation assays both indicated that LINC00261 could inhibit cell proliferation in CRL-7566. Flow cytometry analysis confirmed that LINC00261 mediated inhibition of cell proliferation, which might be a consequence of inducting apoptosis. Furthermore, transwell migration assay indicated that LINC00261 could inhibit cell migration in endometriosis. Conclusion LncRNA LINC00261 is capable of inhibiting cell growth and migration in endometriosis.