研究报告

基于宏基因组学技术解析工业发酵蔬菜中亚硝酸盐形成及降解机理

  • 伍亚龙 ,
  • 杨姗 ,
  • 陈功 ,
  • 张其圣 ,
  • 汪冬冬 ,
  • 唐垚 ,
  • 史梅莓 ,
  • 吕鹏军 ,
  • 王勇
展开
  • 1(四川东坡中国泡菜产业技术研究院,四川 眉山,620000)
    2(四川益动源生物科技有限公司,四川 眉山,620000)
    3(四川省食品发酵工业研究设计院有限公司,四川 成都,611130)
第一作者:学士,高级工程师(王勇工程师为通信作者,E-mail:a1272181492@163.com)

收稿日期: 2023-10-13

  修回日期: 2023-12-06

  网络出版日期: 2024-11-28

Metagenomic analysis of formation and degradation mechanism of nitrite in industrially fermented vegetables

  • WU Yalong ,
  • YANG Shan ,
  • CHEN Gong ,
  • ZHANG Qisheng ,
  • WANG Dongdong ,
  • TANG Yao ,
  • SHI Meimei ,
  • LYU Pengjun ,
  • WANG Yong
Expand
  • 1(Sichuan Dongpo Chinese Paocai Industrial Technology Research Institute, Meishan 620000, China)
    2(Sichuan Eden Biology Technology Co.Ltd., Meishan 620000, China)
    3(Sichuan Food and Fermentation Industry Research & Design Institute Co.Ltd., Chengdu 611130, China)

Received date: 2023-10-13

  Revised date: 2023-12-06

  Online published: 2024-11-28

摘要

蔬菜发酵过程中亚硝酸盐形成及降解机制对于控制亚硝酸盐含量,保证发酵蔬菜食用安全性至关重要。该研究利用宏基因组学方法分析了4种工业发酵蔬菜中的微生物群落分布以及功能基因。物种注释结果表明,发酵蔬菜以乳杆菌属(Lactobacillus)、盐单胞菌属(Halomonas)、枝芽孢杆菌属(Virgibacillus)为优势细菌属,以德巴利酵母属(Debaryomyces)、苏吉雅玛酵母属(Sugiyamaella)、毕赤酵母属(Pichia)和舍弗氏酵母属(Scheffersomyces)为优势真菌属。功能基因分析表明,4种工业发酵蔬菜中亚硝酸盐代谢途径包括反硝化作用、异化硝酸盐还原作用和同化硝酸盐还原作用,硝酸盐还原酶、硝基单加氧酶、亚硝酸盐还原酶、一氧化氮还原酶[细胞色素c]、一氧化二氮还原酶、亚硝酸盐还原酶[NADH]、铁氧还蛋白亚硝酸盐还原酶和亚硝酸盐还原酶[NAD(P)H]是亚硝酸盐代谢过程中起主导作用的酶。微生物与功能基因关联分析表明,费斯莫尔德乳杆菌(Lactobacillus versmoldensis)、咸海鲜盐单胞菌(Halomonas jeotgali)、橙色盐单胞菌(Halomonas lutea)、反硝化盐单胞菌(Halomonas halodenitrificans)、盐反硝化枝芽孢杆菌(Virgibacillus halodenitrificans)等细菌以及米曲霉(Aspergillus oryzae)、尖孢镰刀菌(Fusarium oxysporum)等真菌可能参与反硝化作用、异化硝酸盐还原作用,细菌解淀粉嗜盐碱球菌(Natronococcus amylolyticus)和真菌汉逊德巴利酵母(Debaryomyces hansenii)、食腺嘌呤芽生葡萄孢酵母(Blastobotrys adeninivoran)可能参与同化硝酸盐还原作用,从而促进发酵蔬菜中亚硝酸盐的形成与降解。该研究为探究蔬菜自然发酵过程中亚硝酸盐代谢途径、功能基因以及关联微生物信息提供了新的思路,为发酵蔬菜亚硝酸盐调控提供理论依据。

本文引用格式

伍亚龙 , 杨姗 , 陈功 , 张其圣 , 汪冬冬 , 唐垚 , 史梅莓 , 吕鹏军 , 王勇 . 基于宏基因组学技术解析工业发酵蔬菜中亚硝酸盐形成及降解机理[J]. 食品与发酵工业, 2024 , 50(21) : 60 -67 . DOI: 10.13995/j.cnki.11-1802/ts.037648

Abstract

The high nitrite content in fermented vegetables could cause concerns about food safety.Understanding the process of nitrite formation and degradation during vegetable fermentation is crucial for controlling nitrite levels and ensuring the edibility of fermented vegetables.This study evaluated the microbial community structure and functional genes in four industrially fermented vegetables using metagenomics technology.According to the results of non-redundant protein sequences annotation, the predominant bacterial genera in fermented vegetables were Lactobacillus, Halomonas, and Virgibacillus, whereas the major fungal genera were Debaryomyces, Sugiyamaella, Pichia, and Scheffersomyces.The KEGG annotation results showed that the pathways for nitrite metabolism in four industrially fermented vegetables were denitrification, dissimilatory nitrate reduction, and assimilatory nitrate reduction.The enzymes nitrite reductase, nitric oxide reductase [cytochrome c], nitrous-oxide reductase, nitrite reductase [NADH], ferredoxin-nitrite reductase, and nitrite reductase [NAD(P)H] were the main players in nitrite metabolism.Certain bacteria, such as Lactobacillus versmoldensis, Halomonas jeotgali, Halomonas lutea, Halomonas halodenitrificans, Virgibacillus halodenitrificans, and fungi, such as Aspergillus oryzae and Fusarium oxysporum, may be involved in denitrification and dissimilatory nitrate reduction, according to association analysis of microorganisms and functional genes.The assimilatory nitrate reduction may be facilitated by the bacteria Natronococcus amylolyticus and the fungi Debaryomyces hansenii, Blastobotrys adeninivoran.This study provides new insight for investigating the functional genes, core microbial information, and nitrite metabolic pathways in fermented vegetables in the future.It also provides a theoretical framework for controlling nitrite in fermented vegetables.

参考文献

[1] 杨姗, 王卫, 赵楠, 等.发酵蔬菜色泽形成机制及影响因素研究进展[J].食品科学, 2022, 43(23):269-276.
YANG S, WANG W, ZHAO N, et al.Recent advances in understanding mechanism and influential factors of color formation in fermented vegetables[J].Food Science, 2022, 43(23):269-276.
[2] WANG Z X, SHAO Y Y.Effects of microbial diversity on nitrite concentration in Pao cai, a naturally fermented cabbage product from China[J].Food Microbiology, 2018, 72:185-192.
[3] HUANG Y Y, LIANG M H, ZHAO S, et al.Isolation, expression, and biochemical characterization:Nitrite reductase from Bacillus cereus LJ01[J].RSC Advances, 2020, 10(62):37871-37882.
[4] HUANG T T, WU Z Y, ZHANG W X.Effects of garlic addition on bacterial communities and the conversions of nitrate and nitrite in a simulated pickle fermentation system[J].Food Control, 2020, 113:107215.
[5] HUANG Y Y, JIA X Z, YU J J, et al.Effect of different lactic acid bacteria on nitrite degradation, volatile profiles, and sensory quality in Chinese traditional Paocai[J].LWT, 2021, 147:111597.
[6] YANG X Z, HU W Z, JIANG A L, et al.Effect of salt concentration on quality of Chinese northeast sauerkraut fermented by Leuconostoc mesenteroides and Lactobacillus plantarum[J].Food Bioscience, 2019, 30:100421.
[7] 赵天涛, 陈沛沛, 张晟, 等.异养硝化-好氧反硝化菌氮代谢机理的研究进展[J].重庆理工大学学报(自然科学), 2022(1):194-203.
ZHAO T T, CHEN P P, ZHANG S, et al.Research progress on nitrogen metabolism mechanism of heterotrophic nitrification aerobic denitrification bacteria[J].Journal of Chongqing University of Technology (Natural Science), 2022(1):194-203.
[8] 徐柯, 成林林, 袁美, 等.泡豇豆发酵过程中有机酸变化及对亚硝酸盐降解的影响[J].食品与发酵工业, 2019, 45(17):60-65;72.
XU K, CHENG L L, YUAN M, et al.Changes in organic acids and effects on nitrite degradation during pickled cowpea (Vigna sinensis) fermentation[J].Food and Fermentation Industries, 2019, 45(17):60-65;72.
[9] 魏雯丽, 宫尾茂雄, 吴正云, 等.基于宏转录组学技术解析工业豇豆泡菜发酵过程中活性微生物群落结构变化[J].食品与发酵工业, 2020, 46(10):60-65.
WEI W L, WEI W L, WU Z Y, et al.Analysis of active microbial community structure changes in industrial cowpea pickle fermentation based on meta-transcriptomics technology[J].Food and Fermentation Industries, 2020, 46(10):60-65.
[10] LIU D Q, ZHANG C C, ZHANG J M, et al.Metagenomics reveals the formation mechanism of flavor metabolites during the spontaneous fermentation of potherb mustard (Brassica juncea var.multiceps)[J].Food Research International, 2021, 148:110622.
[11] YU Y Y, LI L, XU Y J, et al.Metagenomics reveals the microbial community responsible for producing biogenic amines during mustard[Brassica juncea (L.)] fermentation[J].Frontiers in Microbiology, 2022, 13:824644.
[12] YU Y Y, LI L, XU Y J, et al.Evaluation of the relationship among biogenic amines, nitrite and microbial diversity in fermented mustard[J].Molecules, 2021, 26(20):6173.
[13] ZHOU Q, ZANG S Z, ZHAO Z N, et al.Dynamic changes of bacterial communities and nitrite character during northeastern Chinese sauerkraut fermentation[J].Food Science and Biotechnology, 2017, 27(1):79-85.
[14] XIA Y J, LIU X F, WANG G Q, et al.Characterization and selection of Lactobacillus brevis starter for nitrite degradation of Chinese pickle[J].Food Control, 2017, 78:126-131.
[15] XIANG W L, ZHANG N D, LU Y, et al.Effect of Weissella cibaria co-inoculation on the quality of Sichuan Pickle fermented by Lactobacillus plantarum[J].LWT, 2020, 121:108975.
[16] 陈功, 唐垚, 赵平, 等.泡菜榨菜亚硝酸盐含量及其N2-NH4+转化机理的研究[J].食品与发酵科技, 2021, 57(2):1-13.
CHEN G, TANG Y, ZHAO P, et al.Study on nitrite content and its N2-NH4+ Transforming theory in Paocai and Zhacai[J].Food and Fermentation Sciences & Technology, 2021, 57(2):1-13.
[17] LI W, LI H, LIU Y D, et al.Salinity-aided selection of progressive onset denitrifiers as a means of providing nitrite for anammox[J].Environmental Science & Technology, 2018, 52(18):10665-10672.
[18] HOSSEINI M, AL-RUBAYE M T S, FAKHARI J, et al.Isolation and characterization of denitrifying halophilic bacteria from Bahr Al-Milh Salt Lake, Karbala, Iraq[J].Journal of Applied Biology & Biotechnology, 2018, 6(4):32-36.
[19] 王奇, 王传明, 周雨, 等.泡菜中微生物菌群的研究进展[J].中国调味品, 2021, 46(9):197-200.
WANG Q, WANG C M, ZHOU Y, et al.Research progress of microbial community in pickles[J].China Condiment, 2021, 46(9):197-200.
[20] LABS, KANEHISA.Kyoto Encyclopedia of Genes and Genomes (KEGG):ko90010[EB/OL].(2023-10-01)[2023-10-10].https://www.genome.jp/pathway/ko00910.
[21] 陈翠翠, 梁艳辉, 祝杰.土壤熏蒸剂专利技术分析[J].现代农药, 2022, 21(4):26-30;36.
CHEN C C, LIANG Y H, ZHU J.Analysis on patent technology of soil fumigation[J].Modern Agrochemicals, 2022, 21(4):26-30;36.
[22] SONG Q Z, ZHAO F K, WANG B B, et al.Metagenomic insights into Chinese northeast Suancai:Predominance and diversity of genes associated with nitrogen metabolism in traditional household Suancai fermentation[J].Food Research International, 2021, 139:109924.
[23] 彭永臻, 钱雯婷, 王琦, 等.基于宏基因组的城市污水处理厂生物脱氮污泥菌群结构分析[J].北京工业大学学报, 2019, 45(1):95-102.
PENG Y Z, QIAN W T, WANG Q, et al.Unraveling microbial structure of activated sludge in a full-scale nitrogen removal plant using metagenomic sequencing[J].Journal of Beijing University of Technology, 2019, 45(1):95-102.
[24] 朱婉瑜, 侍浏洋, 赵维, 等.Zn(II)对好氧反硝化菌Acinetobacter sp.JR-142的代谢活性影响[J].微生物学报, 2022, 62(1):275-290.
ZHU W Y, SHI L Y, ZHAO W, et al.Effects of Zn(Ⅱ) on metabolic activity of aerobic denitrifier Acinetobacter sp.JR-142[J].Acta Microbiologica Sinica, 2022, 62(1):275-290.
[25] 黄雪芹, 左勇, 张强, 等.芽菜中高效降解亚硝酸盐菌株的分离鉴定[J].中国调味品, 2020, 45(7):8-11.
HUANG X Q, ZUO Y, ZHANG Q, et al.Isolation and identification of high-efficiency nitrite-degrading bacteria in sprouts[J].China Condiment, 2020, 45(7):8-11.
[26] 安江波, 刘明健, 刘伟, 等.青贮过程中亚硝酸盐转化途径及影响因素研究进展[J].草地学报, 2023, 31(4):943-951.
AN J B, LIU M J, LIU W, et al.Research progress on nitrite transformation pathway and its influencing factors in silage[J].Acta Agrestia Sinica, 2023, 31(4):943-951.
[27] RUAN Y J, KUMAR AWASTHI M, CAI L, et al.Simultaneous aerobic denitrification and antibiotics degradation by strain Marinobacter hydrocarbonoclasticus RAD-2[J].Bioresource Technology, 2020, 313:123609.
[28] VALENZUELA-ENCINAS C, NERIA-GONZÁLEZ I, ALCÁNTARA-HERNÁNDEZ R J, et al.Phylogenetic analysis of the archaeal community in an alkaline-saline soil of the former Lake Texcoco (Mexico)[J].Extremophiles:Life Under Extreme Conditions, 2008, 12(2):247-254.
[29] KIM M S, ROH S W, BAE J W.Halomonas jeotgali sp.nov., a new moderate halophilic bacterium isolated from a traditional fermented seafood[J].Journal of Microbiology, 2010, 48(3):404-410.
[30] SAKURAI N, ASADA A, MANO S, et al.Tandem and single genes of three membrane-bound nitrate transporters in the nar gene cluster of the moderately halophilic denitrifier, Halomonas halodenitrificans[J].DNA Sequence, 2006, 17(5):363-369.
[31] 高于涵, 贾亚婷, 吕岳骏, 等.微生物厌氧代谢耦合多环芳烃降解研究进展[J].环境科学与技术, 2023, 46(5):67-75.
GAO Y H,JIA Y T,LYU Y J,et al.Research advances in biodegradation of PAHs coupled with microbial anaerobic metabolism[J].Environmental Science & Technology, 2023, 46(5):67-75.
[32] VIGLIOTTA G, DI GIACOMO M, CARATA E, et al.Nitrite metabolism in Debaryomyces hansenii TOB-Y7, a yeast strain involved in tobacco fermentation[J].Applied Microbiology and Biotechnology, 2007, 75(3):633-645.
[33] NAKANISHI Y, ZHOU S M, KIM S W, et al.A eukaryotic copper-containing nitrite reductase derived from a NirK homolog gene of Aspergillus oryzae[J].Bioscience, Biotechnology, and Biochemistry, 2010, 74(5):984-991.
[34] ROHE L, OPPERMANN T, WELL R, et al.Nitrite induced transcription of p450nor during denitrification by Fusarium oxysporum correlates with the production of N2 O with a high 15 N site preference[J].Soil Biology and Biochemistry, 2020, 151:108043.
[35] BÖER E, SCHRÖTER A, BODE R, et al.Characterization and expression analysis of a gene cluster for nitrate assimilation from the yeast Arxula adeninivorans[J].Yeast, 2009, 26(2):83-93.
文章导航

/