食品与发酵工业 >

2024 , Vol. 50 >Issue 16: 93 - 100

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.037359

研究报告

产γ-氨基丁酸航天育种乳酸菌的筛选与发酵剂的制备

  • 王清清 ,
  • 张孟雨 ,
  • 江雨静 ,
  • 杨静贤 ,
  • 钟青萍
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  • (广东省食品质量安全重点实验室,华南农业大学 食品学院,广东 广州,510642)
第一作者：硕士研究生(钟青萍教授为通信作者,E-mail:zhongqp@scau.edu.cn)

收稿日期: 2023-09-12

  修回日期: 2023-10-18

  网络出版日期: 2024-09-19

基金资助

广东省自然科学基金项目(2021A1515011083);国家自然科学基金面上项目(31972046);广东省大学生创新训练计划项目(S202310564099);广东省科技计划项目(2020B1212060059)

收起

Screening of space-breeding lactic acid bacteria for γ-aminobutyric acid production and preparation of fermentation starter

  • WANG Qingqing ,
  • ZHANG Mengyu ,
  • JIANG Yujing ,
  • YANG Jingxian ,
  • ZHONG Qingping
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  • (Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, China)

Received date: 2023-09-12

  Revised date: 2023-10-18

  Online published: 2024-09-19

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摘要

乳酸菌(lactic acid bacteria,LAB)作为产γ-氨基丁酸(γ-aminobutyric acid,GABA)的可食用有益菌,在γ-氨基丁酸生产中作为首选菌种。该研究对乳酸菌进行航天诱变,对113株突变株通过纸层析法初筛、高效液相色谱法复筛,获得2株产GABA较稳定的乳酸菌。为了便于直接投入发酵体系生产,利用Box-Benhnken方法,探究两诱变菌株共培养产GABA的适宜配比,并优化两菌株产GABA的共培养体系的生长条件。结果表明,航天育种后遗传性较稳定的植物乳杆菌L1-51产GABA含量较原始菌株提升了59.58%,乳酸片球菌L21-48比出发菌株产GABA含量提高了9.29%。当发酵温度为37 ℃、pH值为6、植物乳杆菌L1-51和乳酸片球菌L21-48接种比例1∶1、接种量6%时,两菌株共同产GABA含量可达(1.675±0.014) mg/mL。研究结果对航天育种菌株产GABA并利用于发酵行业提供一定的理论基础。

关键词： γ -氨基丁酸; 乳酸菌; 航天育种; 共培养; 条件优化

本文引用格式

王清清 , 张孟雨 , 江雨静 , 杨静贤 , 钟青萍 . 产γ-氨基丁酸航天育种乳酸菌的筛选与发酵剂的制备[J]. 食品与发酵工业, 2024 , 50(16) : 93 -100 . DOI: 10.13995/j.cnki.11-1802/ts.037359

Abstract

Lactic acid bacteria (LAB) are often used in the production of γ-aminobutyric acid (GABA) as edible beneficial bacteria.In this study, space mutagenesis was conducted on LAB, two stable GABA-producing LAB strains were obtained from 113 mutant strains through initial screening by paper chromatography and re-screening by high-performance liquid chromatography (HPLC).To facilitate the application in the fermentation system, the Box-Benhnken method was used to optimize the appropriate inoculation ratio of the two mutated strains and the co-culture conditions for GABA production.Results showed that Lactobacillus plantarum L1-51 was genetically stable after aerospace breeding, and the GABA yield was improved by 59.58% compared with that of the original strain.The GABA yield of Pediococcus acidilactici L21-48 was increased by 9.92% compared with that of the starting strain.When the fermentation temperature was 37 ℃, pH was 6, the inoculation ratio of L.plantarum L1-51 and P.acidilactici L21-48 was 1∶1, and the inoculum volume was 6%, the GABA yield was (1.675±0.014) mg/mL in the co-culture system.The results might provide a theoretical basis for the development and utilization of GABA produced by aerospace breeding strains in fermentation industry.

Key words： γ-aminobutyric acid; lactic acid bacteria; aerospace breeding; co-culture; optimization of conditions

参考文献

[1] SCHWEIZER-SCHUBERT S, GORDON J L, EISENLOHR-MOUL T A, et al.Steroid hormone sensitivity in reproductive mood disorders:On the role of the GABA (A) receptor complex and stress during hormonal transitions[J].Frontiers in Medicine, 2021.DOI:10.3389/FMED.2020.479646.
[2] HEPSOMALI P, GROEGER J A, NISHIHIRA J, et al.Effects of oral gamma-aminobutyric acid (GABA) administration on stress and sleep in humans:A systematic review[J].Frontiers in Neuroscience, 2020,14:923.
[3] 符琼,周文化,黄卫文, 等.发芽糙米生物活性物质及生理功能研究进展[J].食品工业科技, 2019, 40(14):325-333.
FU Q, ZHOU W H, HUANG W W, et al.Research progress on bioactive substances and physiological function of germinated brown rice[J].Science and Technology of Food Industry, 2019, 40(14):325-333.
[4] BOONSTRA E, DE KLEIJN R, COLZATO L S, et al.Neurotransmitters as food supplements:The effects of GABA on brain and behavior[J].Frontiers in Psychology, 2015,6:1520.
[5] DURANTI S, LUGLI G A, MANCABELLI L, et al.Prevalence of antibiotic resistance genes among human gut- derived bifidobacteria[J].Applied and Environmental Microbiology, 2017,83(3):e02894-e02816.
[6] LI X L, SONG Y, MA X Y, et al.Lactobacillus plantarum and Lactobacillus fermentum alone or in combination regulate intestinal flora composition and systemic immunity to alleviate obesity syndrome in high-fat diet rat[J].International Journal of Food Science and Technology, 2018, 53(1):137-146.
[7] JIN X F, CHEN W X, CHEN H M, et al.Combination of Lactobacillus plantarum and Saccharomyces cerevisiae DV10 as starter culture to produce mango slurry:microbiological, chemical parameters and antioxidant activity[J].Molecules, 2019,24(23):4349.
[8] HUNG Y P, LEE C C, LEE J C, et al.The potential of probiotics to eradicate gut carriage of pathogenic or antimicrobial-resistant Enterobacterales[J].Antibiotics, 2021, 10(9):1086.
[9] KORT R, WESTERIK N, MARIELA SERRANO L, et al.A novel consortium of Lactobacillus rhamnosus and Streptococcus thermophilus for increased access to functional fermented foods[J].Microbial Cell Factories, 2015, 14(1):195.
[10] 李向来, 申晓林, 王佳, 等.微生物共培养生产化学品的研究进展[J].合成生物学, 2021,2(6):876-885.
LI X L, SHEN X L, WANG J, et al.Recent advances in biosynthesis of chemicals by microbial co-culture[J].Synthetic Biology Journal, 2021, 2(6):876-885.
[11] CHI Y, WANG X J, LI F, et al.Aerospace technology improves fermentation potential of microorganisms[J].Frontiers in Microbiology, 2022,13:896556.
[12] SENATORE G, MASTROLEO F, LEYS N, et al.Effect of microgravity & space radiation on microbes[J].Future Microbiology, 2018, 13:831-847.
[13] 张彬, 杨涛, 管桂臻, 等.酿酒微生物太空育种及在生产中的应用研究(Ⅱ)芝麻香型白酒酿造功能微生物的太空育种[J].酿酒, 2014,41(6):45-50.
ZHANG B, YANG T, GUAN G Z, et al.Mutagting strains in space of brew microorganisms and application of them in production (II) space mutation breeding of functional microorganisms in sesame flavor liquor[J].Liquor Making, 2014, 41(6):45-50.
[14] 陈志梅, 张宗舟, 薛林贵, 等.航天诱变混菌生抽发酵研究[J].甘肃农业大学学报, 2011, 46(3):104-108.
CHEN Z M, ZHANG Z Z, XUE L G, et al.Mixed fermentation of soy sauce with space mutant strains[J].Journal of Gansu Agricultural University, 2011,46(3):104-108.
[15] 马丽, 魏海霞, 汪倩倩, 等.航天诱变毛霉ZY-3在腐乳生产中的应用研究[J].中国酿造, 2015,34(4):62-65.
MA L, WEI H X, WANG Q Q, et al.Application of space mutation strain Mucor ZY-3 in sufu production[J].China Brewing, 2015,34(4):62-65.
[16] 田蕊. 产γ-氨基丁酸乳酸菌的筛选鉴定、紫外诱变及发酵条件优化[D].呼和浩特:内蒙古农业大学, 2018.
TIAN R. Screening, identification, UV mutagenesis and fermentation conditions optimization of γ-aminobutyric acid by Lactobacillus. Hohhot: Inner Mongolia Agricultural University, 2018.
[17] 马牧然. 产γ-氨基丁酸乳酸菌的筛选及培养条件的优化[D].呼和浩特:内蒙古农业大学, 2021.
MA M R.Screening of lactic acid bacteria producing γ- aminobutyric acid and optimization of culture conditions[D].Hohhot:Inner Mongolia Agricultural University, 2021.
[18] LI H X, PEI J F, WEI C, et al.Sodium-ion-free fermentative production of GABA with Levilactobacillus brevis CD0817[J].Metabolites, 2023, 13(5):608.
[19] LIU H, NI Y, YU Q, et al.Evaluation of co-fermentation of L.plantarum and P.kluyveri of a plant-based fermented beverage:Physicochemical, functional, and sensory properties[J].Food Research International, 2023,172:113060.
[20] 刘玺, 宋照军, 王树宁, 等.富硒发酵香肠的工艺研究[J].食品科学, 2009,30(20):471-474.
LIU X, SONG Z J, WANG S N, et al.Investigation of processing techniques for selenium-enriched fermented sausages[J].Food Science, 2009,30(20):471-474.
[21] 高世阳. 乳酸菌应用榨菜腌制工艺研究[D].杭州:浙江大学, 2014.
GAO S Y. Application of lactic acid bacteria in mustard pickling process. Hangzhou: Zhejiang University, 2014.
[22] KIM J A, PARK M S, KANG S A, et al.Production of γ-aminobutyric acid during fermentation of Gastrodia elata Bl.by co-culture of Lactobacillus brevis GABA 100 with Bifidobacterium bifidum BGN4[J].Food Science and Biotechnology, 2014,23(2):459-466.
[23] SHAO D Y, YAO L B, RIAZ M S, et al.Simulated microgravity affects some biological characteristics of Lactobacillus acidophilus[J].Applied Microbiology and Biotechnology, 2017,101(8):3439-3449.
[24] NEJATI F, RIZZELLO C G, DI CAGNO R, et al.Manufacture of a functional fermented milk enriched of angiotensin-I converting enzyme (ACE)-inhibitory peptides and γ-amino butyric acid (GABA)[J].LWT-Food Science and Technology, 2013,51(1):183-189.
[25] 方颖, 陈依彤, 林咏珊, 等.富含γ-氨基丁酸发酵豆酸乳的研制[J].中国乳品工业, 2021,49(8):60-64.
FANG Y, CHEN Y T, LIN Y S, et al.Research on fermented soybean yogurt rich in γ-aminobutyric acid[J].China Dairy Industry, 2021,49(8):60-64.
[26] KIM N, KIM J M, RA C.Evaluation of γ-aminobutyric acid (GABA) production by lactic acid bacteria using 5-L fermentor[J].Microbiology and Biotechnology Letters, 2021,49(4):559-565.
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