寒地黑土中产γ-氨基丁酸乳酸菌筛选及发酵培养基优化
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摘要
利用MRS培养基分别从牡丹江、漠河、五常的寒地黑土中筛选产γ-氨基丁酸(GABA)的乳酸菌菌株,对其做形态学观察和生理生化鉴定,并进行了发酵培养基组成的优化实验,分析比较不同碳源、氮源、碳氮比、初始p H和L-谷氨酸浓度对产GABA的影响。结果表明:共筛出五株菌株,分别编号为SbO001、SbO002、SbO003、SbO004、SbO005,可以将五株菌株初步鉴定为链球菌科(Streptococcaceae)。当发酵培养基组分碳源为葡萄糖,氮源为硫酸铵,碳氮比1∶1,初始pH为5.5,底物为1%的L-谷氨酸时,菌株产GABA的能力较好,为1.112 g/L。本实验将为寒地黑土中产GABA乳酸菌的筛选提供一定的理论依据。
The lactic acid bacteria producing γ-aminobutyric acid( GABA) strains were screened from the black soil of Mudanjiang,Mohe and Wuchang by MRS medium,and their morphological observation,physiological and biochemical identification were carried out,and the fermentation medium composition was optimized.Experiments were performed to analyze the effects of different carbon sources,nitrogen sources,carbon to nitrogen ratios,initial p H and L-glutamic acid concentration on GABA production. The results showed that five strains were examined,which were numbered SbO001,SbO002,SbO003,SbO004 and SbO005. Five strains could be initially identified as Streptococcaceae.When the carbon source of fermentation medium was glucose,the nitrogen source was ammonium sulfate,the carbon-nitrogen ratio was 1∶ 1,the initial p H was 5.5,and the substrate was 1% l-glutamic acid,the strain had a good gaba-producing capacity of 1.112 g/L.This experiment will provide some theoretical basis for the screening of lactobacillus GABA in the middle black soil of the cold region.
The lactic acid bacteria producing γ-aminobutyric acid( GABA) strains were screened from the black soil of Mudanjiang,Mohe and Wuchang by MRS medium,and their morphological observation,physiological and biochemical identification were carried out,and the fermentation medium composition was optimized.Experiments were performed to analyze the effects of different carbon sources,nitrogen sources,carbon to nitrogen ratios,initial p H and L-glutamic acid concentration on GABA production. The results showed that five strains were examined,which were numbered SbO001,SbO002,SbO003,SbO004 and SbO005. Five strains could be initially identified as Streptococcaceae.When the carbon source of fermentation medium was glucose,the nitrogen source was ammonium sulfate,the carbon-nitrogen ratio was 1∶ 1,the initial p H was 5.5,and the substrate was 1% l-glutamic acid,the strain had a good gaba-producing capacity of 1.112 g/L.This experiment will provide some theoretical basis for the screening of lactobacillus GABA in the middle black soil of the cold region.
引文
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[23]Tajabadi N,Ebrahimpour A,Baradaran A,et al.Optimization ofγ-aminobutyric acid production by Lactobacillus plantarum Taj-Apis362 from honeybees[J]. Molecules,2015,20(4):6654-6669.
[24]李雯.微生物发酵法生产γ-氨基丁酸(GABA)的研究[D].哈尔滨:哈尔滨商业大学,2014.
[25]Haixiang Li,Ting Qiu,Guidong Huang,et al.Production of gamma-aminobutyric acid by Lactobacillus brevis NCL912 using fed-batch fermentation[J].Microbial Cell Factories,2010,9(1):85-85.
[26]姜宏瑛.理化诱变提高乳酸菌STX2γ-氨基丁酸产量的研究[D].南京:南京农业大学,2013.
[27]李鹏.代谢工程改造大肠杆菌生产γ-氨基丁酸[D].杭州:浙江工商大学,2017.
[2]彭俊华,张峰,齐心亮,等.介绍一种大鼠脑中γ-氨基丁酸受体的检测方法[J].西北国防医学杂志,1999,20(4):266-268.
[3]苏国兴,董必慧,刘友良,等.γ-氨基丁酸在高等植物体内的代谢和功能[J].植物生理学报,2003,39(6):670-676.
[4]李海星,江英英,曹郁生.产γ-氨基丁酸乳酸菌的筛选及初步鉴定[J].天然产物研究与开发,2007,19(3):455-510.
[5]Abdou A M,Higashiguchi S,Horie K,et al. Relaxation and immunity enhancement effects ofγ-Aminobutyric acid(GABA)administration in humans[J].Biofactors,2006,26(3):201.
[6]Sang-Min Han,Sun-Jeong Jeon,Hyang-Burm Lee,et al.Screening ofγ-aminobutyric acid(gaba)-producing wild yeasts and their microbiological characteristics[J].The Korean Journal of Mycology,2016,44(2):87-93.
[7]Guido Capitani,Daniela De Biase,Caterina Aurizi,et al.Crystal structure and functional analysis of Escherichia coli glutamate decarboxylase[J]. The EMBO Journal,2014,22(16):4027-4037.
[8]程传兴,刘晓飞,王薇.γ-氨基丁酸的生理功能及制备方法[J].哈尔滨商业大学学报:自然科学版,2017,33(6):693-697.
[9]Sanchart C,Rattanaporn O,Haltrich D,et al. Technological and safety properties of newly isolated GABA-producing Lactobacillus futsaii strains[J]. Journal of Applied Microbiology,2016,121(3):734-745.
[10]BelicováA,MikuláováM,Duinsky R. Probiotic potential and safety properties of Lactobacillus plantarum from Slovak Bryndza cheese[J]. Bio Med research international,2013,2013(2):760298.
[11]梁金钟,田宇,王风青.从酸菜液中筛选产γ-氨基丁酸的菌株[J].食品科学,2011(23):244-249.
[12]韩雪,梁金钟.产γ-氨基丁酸乳酸菌菌株的分离鉴定[J].食品工业科技,2012,33(20):180-183.
[13]刘佳荣,梁金钟.黄浆水中高产γ-氨基丁酸乳酸菌的筛选及鉴定[J].食品科学,2014,35(23):221-225.
[14]徐冬云,周立平,童振宇,等.产γ-氨基丁酸乳酸菌的分离筛选[J].现代食品科技,2006,22(3):59-61.
[15]凌代文,东秀珠.乳酸细菌分类鉴定及实验方法[M].北京:中国轻工业出版社,1993.
[16]邹鹏.产细菌素乳酸菌的分离鉴定及培养条件研究[D].哈尔滨:黑龙江大学,2006.
[17]杨胜远,陆兆新,吕凤霞,等.谷氨酸脱羧酶活力测定中GABA比色定量方法研究[J].食品科学,2006,27(7):205-206.
[18]布坎南,吉布斯.伯杰细菌鉴定手册.8版[M].北京:科学出版社,1984.
[19]田蕊,萨如拉,段晓霞,等.产γ-氨基丁酸乳酸菌的筛选鉴定及其紫外诱变研究[J].食品工业科技,2018,39(17):128-132.
[20]江英英.产γ-氨基丁酸乳酸菌的筛选及其发酵条件优化研究[D].南昌:南昌大学,2007.
[21]冯培勇,赵彦宏,张丽.响应面法优化黑曲霉产纤维素酶发酵条件[J].食品科学,2009,30(23):335-339.
[22]陆小雪.产朴氨基丁酸乳酸菌分离与发酵条件优化及饮料开发[D].南京:南京农业大学,2008:18-20.
[23]Tajabadi N,Ebrahimpour A,Baradaran A,et al.Optimization ofγ-aminobutyric acid production by Lactobacillus plantarum Taj-Apis362 from honeybees[J]. Molecules,2015,20(4):6654-6669.
[24]李雯.微生物发酵法生产γ-氨基丁酸(GABA)的研究[D].哈尔滨:哈尔滨商业大学,2014.
[25]Haixiang Li,Ting Qiu,Guidong Huang,et al.Production of gamma-aminobutyric acid by Lactobacillus brevis NCL912 using fed-batch fermentation[J].Microbial Cell Factories,2010,9(1):85-85.
[26]姜宏瑛.理化诱变提高乳酸菌STX2γ-氨基丁酸产量的研究[D].南京:南京农业大学,2013.
[27]李鹏.代谢工程改造大肠杆菌生产γ-氨基丁酸[D].杭州:浙江工商大学,2017.
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