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      Functional Microbiota for Polypeptide Degradation during Hypertonic Moromi-Fermentation of Pixian Broad Bean Paste

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          Abstract

          Traditional fermented bean pastes are indispensable seasonings in many East Asian countries. They are produced via hypertonic solutions by spontaneous fermentation. Functional, unknown microbiota carry great risks for food safety and stable quality. Thus, analysis and subsequent utilization of functional microbiota will be a good strategy to resolve these problems. During bean fermentation, the microbial functions were divided into two stages, including first stage-raw material (polypeptide) degradation and second stage-amino acid catabolism. In this study, we aimed to analyze the functional microbiota of first stage. Omics-studies, including high-throughput sequencing, correlation analysis and extracellular proteome, were used to generate candidate functional microbes for polypeptide degradation in this study. Then, we cultured the candidate functional microbes. After the batch fermentation and enzymatic analysis, we found three strains secreted peptidase and resulted amino acid accumulation, involving Aspergillus niger, Candida zeylanoides and Bacillus licheniformis. Thus, A. niger, C. zeylanoides and B. licheniformis conducted the functional microbiota for polypeptide degrading during hypertonic moromi fermentation. This study supplies a strategy for functional microbiota analysis. In addition, this is the first report that C. zeylanoides can secrete proteome and produce amino acids from polypeptide.

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          Assessment of soil microbial community structure by use of taxon-specific quantitative PCR assays.

          Here we describe a quantitative PCR-based approach to estimating the relative abundances of major taxonomic groups of bacteria and fungi in soil. Primers were thoroughly tested for specificity, and the method was applied to three distinct soils. The technique provides a rapid and robust index of microbial community structure.
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            Survey of the transcriptome of Aspergillus oryzae via massively parallel mRNA sequencing

            Aspergillus oryzae, an important filamentous fungus used in food fermentation and the enzyme industry, has been shown through genome sequencing and various other tools to have prominent features in its genomic composition. However, the functional complexity of the A. oryzae transcriptome has not yet been fully elucidated. Here, we applied direct high-throughput paired-end RNA-sequencing (RNA-Seq) to the transcriptome of A. oryzae under four different culture conditions. With the high resolution and sensitivity afforded by RNA-Seq, we were able to identify a substantial number of novel transcripts, new exons, untranslated regions, alternative upstream initiation codons and upstream open reading frames, which provide remarkable insight into the A. oryzae transcriptome. We were also able to assess the alternative mRNA isoforms in A. oryzae and found a large number of genes undergoing alternative splicing. Many genes and pathways that might be involved in higher levels of protein production in solid-state culture than in liquid culture were identified by comparing gene expression levels between different cultures. Our analysis indicated that the transcriptome of A. oryzae is much more complex than previously anticipated, and these results may provide a blueprint for further study of the A. oryzae transcriptome.
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              Environmental Microbiota Drives Microbial Succession and Metabolic Profiles during Chinese Liquor Fermentation.

              Many microorganisms in environment participate in the fermentation process of Chinese liquor. However, it is unknown what extent of the environmental microbiota influences on fermentation. In this study, high-throughput sequencing combined with multiphasic metabolite target analysis were applied to study the microbial succession and metabolism changes during Chinese liquor fermentation from two environments (old and new workshops). SourceTracker was applied to evaluate the contribution of environmental microbiota to fermentation. Results showed that Daqu contributed 9.10-27.39% of bacterial communities and 61.06-80.00% of fungal communities to fermentation, whereas environments (outdoor ground, indoor ground, tools and other unknown environments) contributed 62.61-90.90% of bacterial communities and 20.00-38.94% of fungal communities to fermentation. In old workshop, six bacteria [Lactobacillus (11.73% average relative abundance), Bacillus (20.78%), Pseudomonas (6.13%), Kroppenstedtia (10.99%), Weissella (16.64%) and Pantoea (3.40%)] and five fungi [Pichia (55.10%), Candida (1.47%), Aspergillus (10.66%), Saccharomycopsis (22.11%) and Wickerhamomyces (3.35%)] were abundant at the beginning of fermentation. However, in new workshop, the change of environmental microbiota decreased the abundances of Bacillus (5.74%), Weissella (6.64%), Pichia (33.91%), Aspergillus (7.08%) and Wickerhamomyces (0.12%), and increased the abundances of Pseudomonas (17.04%), Kroppenstedtia (13.31%), Pantoea (11.41%), Acinetobacter (3.02%), Candida (16.47%) and Kazachstania (1.31%). Meanwhile, in new workshop, the changes of microbial community resulted in the increase of acetic acid, lactic acid, malic acid and ethyl acetate, and the decrease of ethyl lactate during fermentation. This study showed that environmental microbiota was an important source of fermentation microbiota, and could drive both the microbial succession and the metabolic profiles during liquor fermentation.IMPORTANCE Traditional solid-state fermentation of foods and beverages is mainly carried out by complex microbial communities from raw materials, starters and the processing environments. However, it is still unclear how the environmental microbiota influences on the quality of fermented foods and beverages, especially for Chinese liquor. In this study, we utilized high-throughput sequencing, microbial source tracking and multiphasic metabolite target analysis to analyze the origins of microbiota and the metabolic profiles during liquor fermentation. This study contributes to a deeper understanding of the role of environmental microbiota during fermentation.
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                Author and article information

                Journal
                Foods
                Foods
                foods
                Foods
                MDPI
                2304-8158
                14 July 2020
                July 2020
                : 9
                : 7
                : 930
                Affiliations
                [1 ]Key Laboratory of Industrial Biotechnology of Ministry of Education, State Key Laboratory of Food Science and Technology, School of Biotechnology, Jiangnan University, Wuxi 214100, China; zhanglj@ 123456jiangnan.edu.cn (L.Z.); yidabaojnu@ 123456foxmail.com (Y.B.)
                [2 ]Sichuan Pixian Douban Company Limited, Chengdu 611730, China; chenhaifeng1991@ 123456foxmail.com (H.C.); jiaquanhuang2020@ 123456163.com (J.H.)
                Author notes
                [* ]Correspondence: yxu@ 123456jiangnan.edu.cn ; Tel.: +86-510-8591-8201
                Article
                foods-09-00930
                10.3390/foods9070930
                7404569
                32674449
                5f3fa753-7cac-424c-b960-6b61f15baf00
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 14 June 2020
                : 09 July 2020
                Categories
                Article

                polypeptide degradation,functional microbiota,hypertonic condition,candida,aspergillus

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