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A Gram-stain-negative, coccus-shaped, obligately anaerobic, non-motile and non-spore-forming bacterium, designated strain JN500902 T , was isolated from the mud in a fermentation cellar used continuously over 30 years for Chinese strong-flavour baijiu production. Colonies were white, circular, convex and smooth-edged. Growth was observed at 20-40 °C (optimum, 37 °C), at pH 5.0-10 (optimum, pH 7.5), with 0-2 % (w/v) NaCl and with 0-4 % (v/v) ethanol. The Biolog assay demonstrated positive reactions of strain JN500902 T in the metabolism of l-fucose and pyruvate. The predominant cellular fatty acids (>10 %) consisted of C 16 : 0 and C 14 : 0 . The major end metabolites of strain JN500902 T were acetic acid and ethanol when incubated anaerobically in liquid reinforced clostridial medium. Acetate was the major organic acid end product. The complete genome size of strain JN500902 T was 3 420 321 bp with 3327 identified genes. The G+C content was 43.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain JN500902 T with the family Lachnospiraceae , having low sequence similarity (92.8 %) to the nearest type strain, Syntrophococcus sucromutans DSM 3224 T and forming a clearly distinct branch. Core genome phylogenetic analysis of the isolate and 134 strains belonging to the family Lachnospiraceae also revealed that strain JN500902 T was well-separated from other genera of this family as a monophyletic clade. The average nucleotide identity and amino acid identity values between strain JN500902 T and 134 Lachnospiraceae strains were less than 74 and 65 %, respectively. Considering its polyphasic characteristics, strain JN500902 T represents a novel genus and species within the family Lachnospiraceae , for which the name Novisyntrophococcus fermenticellae gen. nov., sp. nov. is proposed. The type strain is JN500902 T (=CICC 24502 T =JCM 33939 T ).

Humans have used high salinity for the production of bean-based fermented foods over thousands of years. Although high salinity can inhibit the growth of harmful microbes and select functional microbiota in an open environment, it also affects fermentation efficiency of bean-based fermented foods and has a negative impact on people's health. Therefore, it is imperative to develop novel defined starter cultures for reduced-salt fermentation in a sterile environment. Here, we explored the microbial assembly and function in the fermentation of traditional Chinese broad bean paste with 12% salinity. The results revealed that the salinity and microbial interactions together drove the dynamic of community and pointed out that five dominant genera ( Staphylococcus , Bacillus , Weissella , Aspergillus , and Zygosaccharomyces ) may play different key roles in different fermentation stages. Then, core species were isolated from broad bean paste, and their salinity tolerance, interactions, and metabolic characteristics were evaluated. The results provided an opportunity to validate in situ predictions through in vitro dissection of microbial assembly and function. Last, we reconstructed the synthetic microbial community with five strains ( Aspergillus oryzae , Bacillus subtilis , Staphylococcus gallinarum , Weissella confusa , and Zygosaccharomyces rouxii ) under different salinities and realized efficient fermentation of broad bean paste for 6 weeks in a sterile environment with 6% salinity. In general, this work provided a bottom-up approach for the development of a simplified microbial community model with desired functions to improve the fermentation efficiency of bean-based fermented foods by deconstructing and reconstructing the microbial structure and function. IMPORTANCE Humans have mastered high-salinity fermentation techniques for bean-based fermented product preparation over thousands of years. High salinity was used to select the functional microbiota and conducted food fermentation production with unique flavor. Although a high-salinity environment is beneficial for suppressing harmful microbes in the open fermentation environment, the fermentation efficiency of functional microbes is partially inhibited. Therefore, application of defined starter cultures for reduced-salt fermentation in a sterile environment is an alternative approach to improve the fermentation efficiency of bean-based fermented foods and guide the transformation of traditional industry. However, the assembly and function of self-organized microbiota in an open fermentation environment are still unclear. This study provides a comprehensive understanding of microbial function and the mechanism of community succession in a high-salinity environment during the fermentation of broad bean paste so as to reconstruct the microbial community and realize efficient fermentation of broad bean paste in a sterile environment., (Copyright © 2020 American Society for Microbiology.)

Keratinases are promising alternatives over ordinary proteases in several industrial applications due to their unique properties compared with their counterparts in the protease categories. However, their large-scale industrial application is limited by the low expression and poor fermentation efficiency of keratinase. Here, we demonstrate that the expression level of keratinase can be improved by constructing a more efficient enzyme expression system hereby enables the highest production titer as regarding recombinant keratinase production to date. Specially, ten promoters were evaluated and the aprE promoter exhibits a significant promotion of keratinase (kerBv) titer from 165 U/mL to 2605 U/mL in Bacillus subtilis. The batch fermentation mode resulted in a maximum keratinase activity of 7176 U/mL at 36 h in a 5-L fermenter. Furthermore, the extracellular keratinase activity attained up to 16,860 U/mL via fed-batch fermentation within 30 h. The combination of keratinase with l-cysteine brings about 66.4 % degree of degradation of feather. Our work provides a new insight into the development of efficient keratinase fermentation processes with B. subtilis cell factory., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Butyrate and its derivates pertain to the key aroma contributors of strong-flavour baijiu, a kind of Chinese liquors, that is produced from grains by solid-state multispecies anaerobic fermentation in a mud cellar. Microbes inhabiting in the fermentation pit mud largely determines baijiu's flavour and quality. In order to shed light on the microbial functional groups driving butyrate production in pit mud, clone library analysis was firstly performed and the results demonstrated that Clostridia (relative abundance: 50%) and Bacilli (37%) were major groups possessing butyrate kinase (buk) pathway and Clostridia (98%) dominated butyryl-CoA:acetate CoA-transferase (but) pathway. According to Clostridial specific-16S rRNA gene sequencing analysis, we found the resilience character of Clostridial community in pit mud. Amongst Clostridial groups, 32.0% of the sequences were grouped into Clostridiales incertae sedis, followed by Heliobacteriaceae (18.3%) and Clostridiaceae 1 (8.4%). Moreover, Hydrogenispora, Sedimentibacter and Clostridium were the top three abundant genera. Relative abundance of Hydrogenispora was higher in the late days of fermentation, while Sedimentibacter exhibited higher proportion in the early days. Different from the previous studies using universal bacterial primer sets, Hydrogenispora was first reported as one dominant genus in pit mud. As for the reported potential butyrate producer Clostridium, nineteen species were obtained and ten of them were first isolated from the pit mud. Amongst them, buk was identified in eleven species by PCR analysis, while but was identified in the other seven, indicating the species-specific butyrate synthesis pathways of Clostridium. This study provides a perspective on targeting and isolating specific functional microbes in baijiu microbiota with the gene sequence-based medium prediction method., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Over thousands of years, humans have mastered the natural vinegar fermentation technique of cultivating functional microbiota on different raw materials. Functional microbial communities that form reproducibly on non-autoclaved raw materials through repeated batch acetic acid fermentation underpin the flavour development of traditional cereal vinegars. However, how to modulate rationally and optimise the metabolic function of these naturally engineered acidic ecosystems remains unclear. Exploring two key minorities in a vinegar ecosystem, including microbial functions (e.g., flavour and aroma synthesis) and microbial strains, is a crucial step for the vinegar industry to modulate the metabolic function of vinegar microbiota, to monitor the fermentation process, and to maintain the flavour quality of final product., (Copyright © 2017. Published by Elsevier Ltd.)

Multispecies solid-state fermentation (MSSF), a natural fermentation process driven by reproducible microbiota, is an important technique to produce traditional fermented foods. Flavours, skeleton of fermented foods, was mostly produced by microbiota in food ecosystem. However, the association between microbiota and flavours and flavour-producing core microbiota are still poorly understood. Here, acetic acid fermentation (AAF) of Zhenjiang aromatic vinegar was taken as a typical case of MSSF. The structural and functional dynamics of microbiota during AAF process was determined by metagenomics and favour analyses. The dominant bacteria and fungi were identified as Acetobacter, Lactobacillus, Aspergillus, and Alternaria, respectively. Total 88 flavours including 2 sugars, 9 organic acids, 18 amino acids, and 59 volatile flavours were detected during AAF process. O2PLS-based correlation analysis between microbiota succession and flavours dynamics showed bacteria made more contribution to flavour formation than fungi. Seven genera including Acetobacter, Lactobacillus, Enhydrobacter, Lactococcus, Gluconacetobacer, Bacillus and Staphylococcus were determined as functional core microbiota for production of flavours in Zhenjiang aromatic vinegar, based on their dominance and functionality in microbial community. This study provides a perspective for bridging the gap between the phenotype and genotype of ecological system, and advances our understanding of MSSF mechanisms in Zhenjiang aromatic vinegar.

Large-scale submerged fermentation (SmF) of Antrodia camphorata (A. camphorata) usually encounters challenges including tedious preparation of mycelial inoculum, long fermentation period (10-14 d), and poor repeatability. Here we developed an asexual reproduction-based repeated batch fermentation (RBF) process for bioactive metabolites production by A. camphorata ATCC 200183. Compared with traditional batch fermentation, production time was shortened to 58 d from 80 d (overall time for eight cycles) using the RBF process established in this study, and accordingly, the productivities of bioactive metabolites (including antrodins) were improved by 40-60%. Kinetic parameters (α is 2.1-18.7 times as β) indicated that the cell growth was the major contribution for bioactive metabolites production. The RBF shows excellent batch-repeatability (Pearson correlation coefficient of 0.998±0.001), together with advantages of energy-efficient, low cost, and labor-saving, RBF process can be implemented to SmF by other filamentous fungi., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In this study, alteration in morphology of submergedly cultured Antrodia camphorata ATCC 200183 including arthroconidia, mycelia, external and internal structures of pellets was investigated. Two optimization models namely response surface methodology (RSM) and artificial neural network (ANN) were built to optimize the inoculum size and medium components for intracellular triterpenoid production from A. camphorata. Root mean squares error, R (2), and standard error of prediction given by ANN model were 0.31%, 0.99%, and 0.63%, respectively, while RSM model gave 1.02%, 0.98%, and 2.08%, which indicated that fitness and prediction accuracy of ANN model was higher when compared to RSM model. Furthermore, using genetic algorithm (GA), the input space of ANN model was optimized, and maximum triterpenoid production of 62.84 mg l(-1) was obtained at the GA-optimized concentrations of arthroconidia (1.78 × 10⁵ ml(-1)) and medium components (glucose, 25.25 g l(-1); peptone, 4.48 g l(-1); and soybean flour, 2.74 g l(-1)). The triterpenoid production experimentally obtained using the ANN-GA designed medium was 64.79 ± 2.32 mg l(-1) which was in agreement with the predicted value. The same optimization process may be used to optimize many environmental and genetic factors such as temperature and agitation that can also affect the triterpenoid production from A. camphorata and to improve the production of bioactive metabolites from potent medicinal fungi by changing the fermentation parameters.

Collagen is a kind of high macromolecular protein with unique tissue distribution and distinctive functions in the body. At present, most collagen products are extracted from the tissues and organs of mammals or marine fish. However, this method exhibits several disadvantages, including low efficiency and serious waste generation, which makes it difficult to meet the current market demand. With the rapid development of synthetic biology and the deepening of high-density fermentation technology, the collagen preparation by biosynthesis strategy emerges as the times require. Co-expression with the proline hydroxylase gene can solve the problem of non-hydroxylated collagen, but the yield may be affected. Therefore, improving the expression through molecular modification and dynamic regulation of synthesis is an entry point for future research. Due to the defects in certain properties of the natural collagen, modification of properties would be benefit for meeting the requirements of practical application. In this paper, in-depth investigations on recombinant expression, fermentation, and modification studies of collagen are conducted. Also, it summarizes the research progress of collagen in food, medicine, and beauty industry in recent years. Furthermore, the future development trend and application prospect of collagen are discussed, which would provide guidance for its preparation and application. [ABSTRACT FROM AUTHOR]

The fermentation of sauce-flavor Baijiu requires high-temperature Daqu as the starter, which is classified into three types: white, yellow, and black. Recent studies highlighted the variability of the microbiota and metabolome profiles of these three types of Daqu, however, what functions they precisely provide in the sauce-flavor Baijiu brewing process remains unclear. In this study, white, yellow, and black Daqu in five different proportions were utilized as starters in field-scale Baijiu fermentation, with the control group lacking Daqu inoculation. Fermented grains inoculated with 100% white Daqu (47.7 °C) had the highest temperature during stacking fermentation, while 100% black Daqu and control groups were lower than 45 °C. Lactic acid (12.6–22.0 mg/g) was the most abundant organic acid, followed by acetic acid (0.7–5.0 mg/g). qPCR analysis revealed that bacterial biomass decreased during fermentation, while fungal biomass increased. A comparison with the control revealed that Daqu contributed high-temperature resistant microorganisms to the stacking fermentation process, primarily Kroppenstedtia , Virgibacillus , Byssochlamys , Thermoascus , and Thermomyces. Meanwhile, stacking fermentation promoted the enrichment of Acetobacter , Kazachstania , and Leiothecium. Mantel test and co-occurrence network analysis revealed that utilizing different fermentation starters could vary the biotic and abiotic factors that drive microbial community succession. We investigated the flavor of Baijiu further, and found that fermentation with white Daqu enhanced the accumulation of ethyl acetate, isoamyl alcohol, and isobutanol, whereas black Daqu increased the accumulation of ethyl lactate, lactic acid, and furfural. This study contributes to the rational usage of different types of Daqu in the manufacturing of Baijiu. [Display omitted] [ABSTRACT FROM AUTHOR]

Untargeted and targeted metabolomics based on LC-MS/MS was applied here to investigate the changes in metabolites following Lactiplantibacillus plantarum (L. plantarum) fermentation of goji juice. The changes in physicochemical characteristics, free amino acid and organic acid levels were quantified. Total phenols, total flavonoids, and antioxidant activity were increased after these L. plantarum fermentation. 31 primary metabolites were screened out by comparison between two groups using multivariate statistical analysis. L. plantarum consumed reducing sugar and most varieties of amino acids. Gamma amino butyric acid (GABA) and phenylacetaldehyde were significantly accumulated via precursor of glutamate and phenylalanine, respectively. Indole-3-lactic acid (ILA) was the characteristic secondary metabolite after fermentation. There were 6.25 and 4.19 mg/L ILA accumulated after LP-D and LP-F fermentation, respectively, whereas it was not detected in goji juice. The accumulation of tyrosol was strain-dependent. Part of the bitter-taste amino acids, including l -methinoine, l -phenylalanine and tryptophan, were converted into their corresponding acetyl-derivatives, which represented umami or kokumi taste. Overall, fermentation by L. plantarum strains altered the metabolite profile and increased the levels of several biologically active substances of goji juice. These results deepen the understanding of L. plantarum metabolism, and the bioconversion associated with flavor and bioactive compounds unraveled here can be used as references in other plant-based materials for LAB fermentation. [ABSTRACT FROM AUTHOR]

Dissolved oxygen is one of the most important bioprocess parameters that could affect cell growth and product formation, and it is easy to control by changing agitation speed. In this work, the effects of agitation speed on the performance of riboflavin production by recombinant Bacillus subtilis RF1 was investigated in fed-batch fermentation. The lower agitation speed (600 rpm) was beneficial for cell growth and riboflavin biosynthesis in the initial phase of fermentation process. While, during the later phase, higher agitation speed (900 rpm) was favor for cell growth and riboflavin biosynthesis. Thus, a two-stage agitation speed control strategy was proposed based on kinetic analysis, in which the agitation speed was controlled at 600 rpm in the first 26 h and then switched to 900 rpm to maintain high μ for cell growth and high q for riboflavin production during the entire fermentation process. However, it was observed that a sharp increase of agitation speed resulted in an adverse effect on cell growth and riboflavin synthesis within a short time. To avoid this phenomenon, a multi-stage agitation speed control strategy was set up based on the two-stage control strategy, the maximum concentration of riboflavin reached 9.4 g l in 48 h with the yield of 0.051 g g by applying this strategy, which were 20.5 and 21.4 % over the best results controlled by constant agitation speeds. [ABSTRACT FROM AUTHOR]

Cultivation in glycerol instead of sugars inhibits 2,3-butanediol (2,3-BD) production by Bacillus amyloliquefaciens. In this study, we report that B. amyloliquefaciens readily produces 2,3-BD from biodiesel-derived glycerol in the presence of beet molasses as a co-substrate. Unexpectedly, the molasses stimulated 2,3-BD production and simultaneously reduced the duration of fermentation. Productivity of 2,3-BD was enhanced at the start of fermentation, and yields increased under continuous molasses supply. Subsequently, 2,3-BD production in molasses-supplemented fed-batch culture was observed. Prior to inoculation of fed-batch fermentation culture, 15 g/l of molasses was added to the bioreactor. After 6 h of incubation, the bioreactor was fed with a solution containing 80 % glycerol and 15 % molasses. The 2,3-BD concentration, yield, and productivity significantly improved, reaching 83.3 g/l, 0.42 g/g, and 0.87 g/l·h, respectively. To our knowledge, these results are the highest report for 2,3-BD fermentation from biodiesel-derived glycerol. [ABSTRACT FROM AUTHOR]

Antrodia camphorata is a well-known Chinese medicinal mushroom that protects against diverse health-related conditions. Submerged fermentation of A. camphorata is an alternative choice for the effective production of bioactive metabolites, but the effects of nutrition and environment on mycelial morphology are largely unknown. In this study, we show that A. camphorata American Type Culture Collection 200183 can form arthrospores in the end of liquid fermentation. Different morphologies of A. camphorata in submerged culture were analyzed using scanning electron microscopy. The optimal carbon and nitrogen sources for sporulation were soluble starch and yeast extract. We found that a carbon-to-nitrogen ratio (C/N) of 40:1, MgSO (0.5 g/l), KHPO (3.0 g/l), an initial pH 5.0, and an inoculum size of 1.5 × 10 spores/ml led to maximum production of arthroconidia. Our results will be useful in the regulation and optimization of A. camphorata cultures for efficient production of arthroconidia in submerged culture, which can be used as inocula in subsequent fermentation processes. [ABSTRACT FROM AUTHOR]

The main purpose of this study was to determine the volatile composition of Zhenjiang aromatic vinegar, one of the four famous China-style cereal vinegars, by using headspace solid-phase microextraction (HS-SPME)/gas chromatography-mass spectrum (GC-MS) and chemometrics. For this purpose, the HS-SPME sampling method for the volatile compounds of Zhenjiang aromatic vinegar was optimized by a second-order rotatable central composite experimental design (CCD). A HS extraction of the volatile compounds by incubation on a 65 µm thickness polydimethylsiloxane/divinylbenzene (PDMS/DVB) fibre during 44.2 min at 69.5°C with 1.9 g NaCl add gave the most effective and accurate extraction. By the optimized method, a total of 58 volatile compounds, including 9 alcohols, 13 acids, 16 esters, 5 aldehydes, 4 ketones and 8 heterocycle compounds, were identified from 13 aromatic vinegar samples manufactured in Zhenjiang region. By principal components analysis (PCA), the thirteen vinegar samples were classified into 3 groups, and 10 volatile compounds were chosen as characteristic compounds of Zhenjiang aromatic vinegars. Copyright © 2012 The Institute of Brewing & Distilling [ABSTRACT FROM AUTHOR]

AbstractVitamin K2, also known as methylnaphthoquinone, is a crucial fat-soluble nutrient necessary for the human body. The biological production of Vitamin K2 has received widespread attention due to its environmental friendliness and maneuverability in recent years. This review provides insights into the modular metabolic pathways of Vitamin K2, lays the foundation for microbial metabolic flow balancing, cofactor engineering and dynamic regulation, and realizes the production of Vitamin K2 by synthesizing artificial cells from scratch. With the intensive development of modern fermentation technology, methods for the preparation of Vitamin K2 using the fermentation strategies of co-culturing and biofilm reactors have emerged. In prokaryotes, the introduction of heptenyl pyrophosphate synthase (HepPPS) and mevalonate acid (MVA) pathway solved the problem of insufficient precursors for Vitamin K2 production but still did not meet the market demand. Therefore, enhancing expression through multi-combinatorial metabolic regulation and innovative membrane reactors is an entry point for future research. Due to the light-induced decomposition and water-insoluble nature of Vitamin K2, the secretion regulation and purification processing also need to be considered in the actual production. Also, it summarizes the research progress of Vitamin K2 in the food and pharmaceutical fields. Additionally, the future development trend and application prospect of Vitamin K2 are also discussed to provide guidance for Vitamin K2 biosynthesis and application. [ABSTRACT FROM AUTHOR]

Hyaluronate lyases (HA lyases) have been proved to distribute widely among microorganisms, with large potential in hyaluronan processing. Here, a highly active HA lyase HylC from Citrobacter freundii strain Cf1 is reported. HylC was expressed in Escherichia coli BL21(DE3) under the regulation of T7 promoter, and purified to electrophoretic homogeneity for enzymatic characterization, which suggested its suitable thermo- and pH stability under 45 °C and pH rang of 4–8, and high halotolerancy in 1.5 M NaCl. The enzyme exhibited the optimal activity under 37 °C and pH 5.5, and was activated by Ca2+, K+, Zn2+, Ni2+ and Li+. Analysis of degradation product proved it cleave HA in endolytic manner, releasing unsaturated disaccharides as final product. Then, through optimization of promoter and construction of dual promoter, expression level of HylC improved from 1.10 × 104 U/mL to 2.64 × 104 U/mL on shake-flask level. Finally, through batch fermentation, a highest activity of 2.65 × 10 5 U/mL was achieved in a 5-L fermenter. Taken together, this work demonstrates the potential of HylC and its recombinant strain in industrial applications. To our knowledge, the HA lyase production reported in this study was the highest level in literatures to date. • A highly active hyaluronate lyase was found, cloned and characterized. • Promoter engineering enhanced recombinant expression of hyaluronate lyase by 2.4-fold. • (3)The highest level of recombinant hyaluronate lyase reached up to 2.65 × 105 U/mL. [ABSTRACT FROM AUTHOR]

[Display omitted] • The richness and diversity of yeast community increased throughout fermentation. • The vast majority of yeasts in the fermentation sourced from the brewing environment. • Wickerhamomyces (29.6 %), Saccharomycopsis (25.0 %), and Torulaspora (14.9 %) were the predominant genera. • These predominant genera showed distinct spatial distributions throughout the fermentation stack. • T. delbrueckii ME22, S. fibuligera ME8, and W. anomalus ME57 drove the formation of floral, fruity, and sweet flavor compounds. Yeasts play a crucial role in determining the quality and yield of sauce-flavor Baijiu, yet the source, succession, and metabolic functions of the yeast community in fermented grains during stacking fermentation remains unclear. In this study, amplicon sequencing combined with solid-state fermentation was used to investigate the structure and function of yeast community during the first-round fermentation of sauce-flavor Baijiu. The richness and diversity of yeast community increased throughout fermentation, with 83.05 % of yeast ASV sourced from the fermentation environment. Fourteen yeast genera were identified, with Wickerhamomyces (29.6 %), Saccharomycopsis (25.0 %), and Torulaspora (14.9 %) being the predominant genera. These genera showed distinct spatial distributions throughout the fermentation stack. Spearman correlation analysis indicated positive correlations between the three genera and multiple volatiles in fermented grains alcohols and esters. After solid-state fermentation in pure culture, T. delbrueckii ME22, S. fibuligera ME8, and W. anomalus ME57 produced distinct floral, fruity, and sweet flavor compounds, such as phenylethyl alcohol, isoamyl alcohol, ethyl acetate, phenethyl acetate, and isoamyl acetate. T. delbrueckii ME22 demonstrated a great capacity for cellulose degradation, whereas S. fibuligera ME8 exhibited enhanced capabilities for protein and starch degradation. This study provides a theoretical reference for the application of yeast in the fermentation of sauce-flavor Baijiu. [ABSTRACT FROM AUTHOR]

Trehalase has attracted widespread attention in medicine, agriculture, food, and ethanol industry due to its ability to specifically degrade trehalose. Efficient expression of trehalase remains a challenge. In this study, a putative trehalase-encoding gene (Tre-zm) from Zunongwangia mangrovi was explored using gene-mining strategy and heterologously expressed in E. coli. Trehalase activity reached 3374 U·mL−1 after fermentation optimization. The scale-up fermentation in a 15 L fermenter was achieved with a trehalase production of 15,068 U·mL−1. The recombinant trehalase TreZM was purified and characterized. It displayed optimal activity at 35 °C and pH 8.5, with Mn2+, Sn2+, Na+, and Fe2+ promoting the activity. Notably, TreZM showed significant inhibition effect on biofilm forming of Staphylococcus epidermidis. The combination of TreZM with a low concentration of antibiotics could inhibit 70 % biofilm formation of Staphylococcus epidermidis and 28 % of Pseudomonas aeruginosa. Hence, this study provides a promising candidate for industrial production of trehalase and highlights its potential application to control harmful biofilms. Gene mining, expression, fermentation, purification, characterization and application of trehalase. [Display omitted] [ABSTRACT FROM AUTHOR]

As the saccharifying and fermentative agent, medium-temperature Daqu (MT-Daqu) plays an irreplaceable role in the production of strong-flavor Baijiu. Numerous studies have focused on the microbial community structure and potential functional microorganisms, however, little is known about the succession of active microbial community and the formation mechanism of community function during MT-Daqu fermentation. In this study, we presented an integrated analysis of metagenomics, metatranscriptomics, and metabonomics covering the whole fermentation process of MT-Daqu to reveal the active microorganisms and their participations in metabolic networks. The results showed that dynamic of metabolites were time-specific, and the metabolites and co-expressed active unigenes were further classified into four clusters according to their accumulation patterns, with members within each cluster displaying a uniform and clear pattern of abundance across fermentation. Based on KEGG enrichment analysis in co-expression clusters and succession of active microbial community, we revealed that Limosilactobacillus , Staphylococcus , Pichia , Rhizopus , and Lichtheimia were metabolically active members at the early stage, and their metabolic activities were conducive to releasing abundant energy to drive multiple basal metabolisms such as carbohydrates and amino acids. Thereafter, during the high temperature period and at the end of fermentation, multiple heat-resistant filamentous fungi were transcriptionally active populations, and they acted as both the saccharifying agents and flavor compound producers, especially aromatic compounds, suggesting their crucial contribution to enzymatic activity and aroma of mature MT-Daqu. Our findings revealed the succession and metabolic functions of the active microbial community, providing a deeper understanding of their contribution to MT-Daqu ecosystem. • The accumulation of metabolites during fermentation was time-specific and staged. • Metabolites and active genes exhibited consistent and clear abundance patterns. • Fungi gradually replaced bacteria as the active species during fermentation. • Lactobacillales, Mucorales, and Pichia initially cooperated in the hydrolyses. • Filamentous fungi contributed to form flavor substances under thermal conditions. [ABSTRACT FROM AUTHOR]

Chinese have learned to regulate salinity for broad bean paste-meju preparation over hundreds of years, yet the mechanisms of salinity in regulating the aroma and safety have not been identified. Here, we explored the effects of salt concentration (6%, 9%, 12%, and 15%, w/w) on the broad bean paste-meju fermentation. Results showed that low salt meju fermented rapidly, flavors such as OAs and FFAs were produced more quickly, and monosaccharides were consumed much more quickly by microorganisms. There was abundant linoleic acid, oleic acid, palmitic acid in fermented meju. Salt concentration resulted in an entirely different flavor of meju. Some acids and phenols compounds were prominent in the low salt group. Whereas the compounds that contributed to sauce aromas such as ethanol, hexadecanoic acid ethyl ester, and 4-ethyl-guaiacol were mainly detected in the high salt group. On the other hand, the low salt condition is beneficial for BAs production, contents of BAs in the 6% salt content broad bean paste-meju was much higher than in other groups (reached 2599.18 ± 725.75 mg/kg). In brief, salt plays an important role in broad bean paste-meju fermentation. Through this study, we would recommend 9%-12%, especially 12% as appropriate salt content for broad bean paste-meju fermentation. • High salt meju is beneficial for the production of sauce aroma. • Monosaccharides accumulated in high salt groups. • Low salt condition is beneficial for the production of BAs. • Tetragenococcus didn't produce a high amount of BAs. [ABSTRACT FROM AUTHOR]

[Display omitted] • The profiles of microbiome and metabolome were explored during the whole fermentation of Zhenjiang aromatic vinegar. • Positive correlations were identified between fungi and lots of flavor metabolites. • Sealed fermentation promoted the accumulation of various flavor compounds. • Jiuqu and seed pei provided microorganisms to initiate starch saccharification and acetic acid fermentation, respectively. • Maiqu was mainly the donor of enzymes in alcohol fermentation. Microbial ecosystems of fermented foods are largely interfered by human activities in myriad ways. The aim of this study was to illuminate the impacts of various starters and environmental variables on the fermentation process of Zhenjiang aromatic vinegar (ZAV), one of the four representative cereal vinegars in China. The effects of environmental variables (e.g., ethanol, total acidity, temperature) and starters (e.g., jiuqu, maiqu, seed pei) on the profiles of microbiome and metabolome (e.g., organic acids, amino acids and volatiles) during fermentation process of ZAV were analyzed. Amongst the four fermentation stages, acetic acid fermentation was the main stage for the accumulation of flavor substances, and subsequently, the contents of acids (mainly acetic, lactic and citric acids) and volatile metabolites (e.g., 2,3-butanedione, acetoin, etc.) continued to enrich in sealed fermentation stage. Principal coordinate analysis (PCoA) and analysis of similarities (ANOSIM) showed that the fungal and bacterial community structures of four fermentation stages were significantly different. As for bacterial community, the dominant OTUs with average relative abundance over 10% in at least one fermentation stage were assigned to the genera Acetilactobacillus , Acetobacter , Acinetobacter , Aeromonas , Lactobacillus , and Pseudomonas. The dominant fungal populations in each fermentation stage were obviously divergent, including Wickerhamomyces , Saccharomyces , Alternaria , Fusarium , etc. SourceTracker analysis demonstrated that jiuqu and seed pei provided microorganisms to initiate starch saccharification and acetic acid fermentation stages, respectively, and maiqu was mainly the donor of enzymes in alcohol fermentation. Spearman correlation coefficients revealed positive relationships between fungal community and various flavor metabolites, indicating the essential role of fungi in the flavor formation of ZAV. This study systematically reveals the effects of fermentation starters and environmental variables on vinegar production and deepens the understanding of the traditional production craft. [ABSTRACT FROM AUTHOR]

Daqu, a brick-shaped product spontaneously fermented under an open environment, has been regarded as the starter of fermentation, raw enzyme preparation and raw materials for baijiu production. However, its contribution in baijiu fermentation has not been fully elaborated yet. Here, the effects of daqu microbiota on baijiu fermentation were investigated under both field-scale and lab-scale conditions. In field-scale baijiu fermentation, the dominant daqu microbes (average relative abundance>10.0%), including unclassified_ Leuconostocaceae , Thermoascus, and Thermomyces , tended to dominate the early stage (0–7 d). However, the rare daqu microbes (average relative abundance <0.1%, e.g. , Kazachstania) tended to dominate the middle and late stages (11–40 d). In addition, some genera showed differences in species diversity between daqu and fermented grains. The average relative abundance of Lactobacillus was over 75% during baijiu fermentation, and most of them were affiliated with Lactobacillus acetotolerans , while Lactobacillus crustorum dominated the Lactobacillus OTUs in daqu. The similar patterns were also observed during lab-scale baijiu fermentation. The results of function prediction showed the enriched metabolic pathways were associated with glycolysis and long-chain fatty acid esters in baijiu fermentation. These results improved the understanding of daqu microbiota function during baijiu fermentation and provided a basic theory to support the regulation of baijiu production. [Display omitted] • Microbiota succession of daqu during alcohol fermentation were studied. • The dominant species of Lactobacillus in fermented grains were different from daqu. • The dominant and rare daqu microbes respectively dominated the early and late stages of alcohol fermentation. [ABSTRACT FROM AUTHOR]