Effect of co-culture of Clostridium strains and Novisyntrophococcus fermenticellae, isolated from pit mud, on growth and short-chain fatty acid metabolism

  • SUN Hong ,
  • CHAI Lijuan ,
  • FANG Guanyu ,
  • LU Zhenming ,
  • ZHANG Xiaojuan ,
  • WANG Songtao ,
  • SHEN Caihong ,
  • SHI Jinsong ,
  • XU Zhenghong
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  • 1(School of Biotechnology, Jiangnan University, Wuxi 214122, China)
    2(National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China)
    3(School of Life Science and Health Engineering, Jiangnan University, Wuxi 214122, China)
    4(Jiangsu Provincial Engineering Research Center for Bioactive Product Processing, Jiangnan University, Wuxi 214122, China)
    5(National Engineering Research Center of Solid-State Brewing, Luzhou 646000, China)

Received date: 2022-05-11

  Revised date: 2022-05-19

  Online published: 2022-09-02

Abstract

To study the interaction between Clostridium, the important functional microorganisms in pit mud of Chinese strong-flavor Baijiu, and other microorganisms in pit mud, we analyzed the differences of genomic functions and short chain fatty acid (SCFA) metabolic pathways of Clostridium species based on comparative genomics.Besides, we selected Novisyntrophococcus fermenticellae JN500902 (N.902) and seven strains of Clostridium isolated from pit mud as the research object, constructed a two-strain coculture system of N.902 and Clostridium strain, and compared the changes of growth and SCFA metabolism in mono- and co-culture.Comparative genomics analysis showed that Clostridium’s SCFA metabolic pathway was interspecific differences.Co-culture experiment results further showed that N.902 had a significant positive effect on the growth and metabolism of Clostridium fermenticellae JN500901 (C.901) and C.luticellarii Clu07, the consumption of glucose and growth increased in coculture.Besides, the accumulation of butyric acid and caproic acid in C.901 increased by about 0.44 folds and 0.63 folds, respectively, and the yield of butyric acid and valeric acid in Clu07 increased by 0.77 times and 2.63 times, respectively.N.902 co-cultured with two valeric acid producing Clostridium strains had no significant effect on the growth, while it promoted the production of valeric acid in C.scatologenes Cls01 and inhibited the accumulation of valeric acid in C.aciditolerans Claci01.Co-culture of N.902 and three butyric acid-producing Clostridium strains, C.beijerinckii Clb01, C.guangxiense Clgx01 and C.tyrobutyricum Clt01 showed higher growth rate and biomass than that of mono-culture in the early stage (before 24 h), but the accumulation of butyric acid decreased in varying degrees at the end of co-culture fermentation.The research on the interaction between Clostridium and N.902 could broaden our understanding of pit mud microbial interaction for aroma production.

Cite this article

SUN Hong , CHAI Lijuan , FANG Guanyu , LU Zhenming , ZHANG Xiaojuan , WANG Songtao , SHEN Caihong , SHI Jinsong , XU Zhenghong . Effect of co-culture of Clostridium strains and Novisyntrophococcus fermenticellae, isolated from pit mud, on growth and short-chain fatty acid metabolism[J]. Food and Fermentation Industries, 2022 , 48(15) : 24 -32 . DOI: 10.13995/j.cnki.11-1802/ts.032308

References

[1] CHAI L J, QIAN W, ZHONG X Z, et al.Mining the factors driving the evolution of the pit mud microbiome under the impact of long-term production of strong-flavor Baijiu[J].Applied and Environmental Microbiology, 2021, 87(17):e0088521.
[2] FU J X, CHEN L, YANG S Z, et al.Metagenome and analysis of metabolic potential of the microbial community in pit mud used for Chinese strong-flavor liquor production[J].Food Research International, 2021, 143:110294.
[3] LIU M K, TANG Y M, GUO X J, et al.Deep sequencing reveals high bacterial diversity and phylogenetic novelty in pit mud from Luzhou Laojiao cellars for Chinese strong-flavor Baijiu[J].Food Research International, 2017, 102:68-76.
[4] HU X L, DU H, REN C, et al.Illuminating anaerobic microbial community and cooccurrence patterns across a quality gradient in Chinese liquor fermentation pit muds[J].Applied and Environmental Microbiology, 2016, 82(8):2 506-2 515.
[5] LIU M K, TANG Y M, ZHAO K, et al.Contrasting bacterial community structure in artificial pit mud-starter cultures of different qualities:A complex biological mixture for Chinese strong-flavor Baijiu production[J].3 Biotech, 2019, 9(3):89.
[6] TAN G X, ZHOU R, ZHANG W Q, et al.Detection of viable and total bacterial community in the pit mud of Chinese strong-flavor liquor using propidium monoazide combined with quantitative PCR and 16S rRNA gene sequencing[J].Frontiers in Microbiology, 2020, 11:896.
[7] ZHENG X W, HAN B Z.Baijiu, Chinese liquor:History, classification and manufacture[J].Journal of Ethnic Foods, 2016, 3(1):19-25.
[8] CHAI L J, XU P X, QIAN W, et al.Profiling the Clostridia with butyrate-producing potential in the mud of Chinese liquor fermentation cellar[J].International Journal of Food Microbiology, 2019, 297:41-50.
[9] 钱玮, 陆震鸣, 柴丽娟, 等.泸型酒酒醅与窖泥中梭菌群落结构、演替和功能差异[J].生物工程学报, 2020, 36(6):1 190-1 197.
QIAN W, LU Z M, CHAI L J, et al.Differences of the structure, succession and function of Clostridial communities between Jiupei and pit mud during Luzhou-flavour Baijiu fermentation[J].Chinese Journal of Biotechnology, 2020, 36(6):1 190-1 197.
[10] ZOU W, YE G B, ZHANG K Z.Diversity, function, and application of Clostridium in Chinese strong flavor Baijiu ecosystem:A review[J].Journal of Food Science, 2018, 83(5):1 193-1 199.
[11] ZHENG Y, HU X L, JIA Z J, et al.Co-occurrence patterns among prokaryotes across an age gradient in pit mud of Chinese strong-flavor liquor[J].Canadian Journal of Microbiology, 2020, 66(9):495-504.
[12] ZHANG Y C, KASTMAN E K, GUASTO J S, et al.Fungal networks shape dynamics of bacterial dispersal and community assembly in cheese rind microbiomes[J].Nature Communications, 2018, 9:336.
[13] BLASCHE S, KIM Y, MARS R A T, et al.Metabolic cooperation and spatiotemporal niche partitioning in a kefir microbial community[J].Nature Microbiology, 2021, 6(2):196-208.
[14] WANG S J, TANG H Z, PENG F, et al.Metabolite-based mutualism enhances hydrogen production in a two-species microbial consortium[J].Communications Biology, 2019, 2:82.
[15] 柴丽娟, 钱玮, 钟小忠, 等.浓香型白酒发酵过程中窖内古菌群落分布特征[J].生物工程学报, 2020, 36(12):2 635-2 643.
CHAI L J, QIAN W, ZHONG X Z, et al.Distribution of archaeal community in the mud pit during strong-flavor Baijiu fermentation[J].Chinese Journal of Biotechnology, 2020, 36(12):2 635-2 643.
[16] DU H, LIU B, WANG X S, et al.Exploring the microbial origins of p-cresol and its co-occurrence pattern in the Chinese liquor-making process[J].International Journal of Food Microbiology, 2017, 260:27-35.
[17] RÖTTJERS L, FAUST K.From hairballs to hypotheses-biological insights from microbial networks[J].FEMS Microbiology Reviews, 2018, 42(6):761-780.
[18] HU X L, DU H, XU Y.Identification and quantification of the caproic acid-producing bacterium Clostridium kluyveri in the fermentation of pit mud used for Chinese strong-aroma type liquor production[J].International Journal of Food Microbiology, 2015, 214:116-122.
[19] XU J L, SUN L P, XING X, et al.Culturing bacteria from fermentation pit muds of Baijiu with culturomics and amplicon-based metagenomic approaches[J].Frontiers in Microbiology, 2020, 11:1223.
[20] CHAI L J, FANG G Y, XU P X, et al.Novisyntrophococcus fermenticellae gen.nov., sp.nov., isolated from an anaerobic fermentation cellar of Chinese strong-flavour Baijiu[J].International Journal of Systematic and Evolutionary Microbiology, 2021, 71(9):DOI:10.1099/ijsem.0.004991.
[21] KUMAR S, STECHER G, LI M, et al.MEGA X:Molecular evolutionary genetics analysis across computing platforms[J].Molecular Biology and Evolution, 2018, 35(6):1 547-1 549.
[22] FELSENSTEIN J.Confidence limits on phylogenies:An approach using the bootstrap[J].Evolution;International Journal of Organic Evolution, 1985, 39(4):783-791.
[23] SEEMANN T.Prokka:rapid prokaryotic genome annotation[J].Bioinformatics, 2014, 30(14):2 068-2 069.
[24] HUERTA-CEPAS J, FORSLUND K, COELHO L P, et al.Fast genome-wide functional annotation through orthology assignment by eggNOG-mapper[J].Molecular Biology and Evolution, 2017, 34(8):2 115-2 122.
[25] ZHAO Y B, JIA X M, YANG J H, et al.PanGP:A tool for quickly analyzing bacterial pan-genome profile[J].Bioinformatics, 2014, 30(9):1 297-1 299.
[26] CHARUBIN K, PAPOUTSAKIS E T.Direct cell-to-cell exchange of matter in a synthetic Clostridium syntrophy enables CO2 fixation, superior metabolite yields, and an expanded metabolic space[J].Metabolic Engineering, 2019, 52:9-19.
[27] ZOU W, YE G B, ZHANG K Z, et al.Analysis of the core genome and pangenome of Clostridium butyricum[J].Genome, 2021, 64(1):51-61.
[28] ZOU W, YE G B, LIU C J, et al.Comparative genome analysis of Clostridium beijerinckii strains isolated from pit mud of Chinese strong flavor Baijiu ecosystem[J].G3 Genes|Genomes|Genetics, 2021, 11(11):jkab317.
[29] UDAONDO Z, DUQUE E, RAMOS J L.The pangenome of the genus Clostridium[J].Environmental Microbiology, 2017, 19(7):2 588-2 603.
[30] 刘昊鹏, 刘超, 王雯, 等.基于厌氧微生物的碳链延长合成高价值化学品反应机理及研究进展:不同电子供体[J].北京化工大学学报(自然科学版), 2020, 47(5):1-17.
LIU H P, LIU C, WANG W, et al.Advances in understanding the mechanism of chain elongation with anaerobic microbes for the synthesis of high value-added chemicals:The effect of different electron donors[J].Journal of Beijing University of Chemical Technology (Natural Science Edition), 2020, 47(5):1-17.
[31] YAO Y, FU B, HAN D F, et al.Formate-dependent acetogenic utilization of glucose by the fecal acetogen Clostridium bovifaecis[J].Applied and Environmental Microbiology, 2020, 86(23):e01870-e01820.
[32] SAN-VALERO P, ABUBACKAR H N, VEIGA M C, et al.Effect of pH, yeast extract and inorganic carbon on chain elongation for hexanoic acid production[J].Bioresource Technology, 2020, 300:122659.
[33] SAN-VALERO P, FERNÁNDEZ-NAVEIRA Á, VEIGA M C, et al.Influence of electron acceptors on hexanoic acid production by Clostridium kluyveri[J].Journal of Environmental Management, 2019, 242:515-521.
[34] DUBER A, ZAGRODNIK R, CHWIALKOWSKA J, et al.Evaluation of the feed composition for an effective medium chain carboxylic acid production in an open culture fermentation[J].Science of the Total Environment, 2020, 728:138814.
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