产γ-氨基丁酸乳酸菌CICC 6238的诱变选育及益生特性分析
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摘要
从中国工业微生物菌种资源库优选的38株乳酸菌中,筛选得到1株γ-氨基丁酸(γ-aminobutyric acid,GABA)产量较高的植物乳杆菌CICC 6238。通过紫外诱变育种,CICC 6238的GABA产量提高至(1.196±0.033)g/L,是原菌株的2.33倍。与商业益生菌嗜酸乳杆菌NCFM和鼠李糖乳杆菌HN001相比,CICC 6238对胆盐、人工胃液和人工肠液具有良好的耐受性。CICC 6238耐胆盐存活率为(36.90±1.63)%,显著高于NCFM[(15.16±1.73)%];在人工胃液消化3 h的存活率为(84.05±1.87)%,显著高于HN001[(61.97±2.45)%];在人工肠液消化6 h后的存活率为(61.50±1.94)%,略低于HN001[(77.44±3.08)%]和NCFM[(79.42±2.08)%]。CICC 6238对4种肠道致病菌尤其是福氏志贺氏菌具有明显的抑制作用,抑菌圈可达(28.31±0.27)mm。CICC 6238既能高产GABA,又具有潜在的益生特性,可作为功能益生菌,应用于未来功能性食品及保健食品的研究开发。
Lactobacillus plantarum CICC 6238 was screened out for γ-aminobutyric acid( GABA) production from 38 lactic acid bacteria deposited in the China Center of Industrial Culture Collection( CICC). The high-yield GABA strains were obtained by ultraviolet mutagenesis,and its production of GABA was(1. 196 ± 0. 033) g/L,which was 2. 33 times as much as original strain. CICC 6238 was studied extensively for its high tolerance to simulated gastric,intestine juices and bile salts with commercial probiotic strains( Lactobacillus acidophilus NCFM and Lactobacillus rhamnosus HN001) as positive control. After incubation in bile salt environment,the survival rate of CICC 6238 was(36. 90 ± 1. 63) % significantly higher than NCFM [( 15. 16 ± 1. 73) %]. After digestion in artificial gastric juice for 3 hours,the survival rate of CICC6238 was(84. 05 ± 1. 87) %,which was remarkably greater than that of HN001 [(61. 97 ± 2. 45) %]. After incubation in artificial intestine juices for 6 hours,the survival rate of CICC6238 was(61. 50 ± 1. 94) %,which was slightly lower than that of HN001 [(77. 44 ± 3. 08) %]and NCFM [(79.42 ± 2. 08) %]. And CICC 6238 also could inhibit the growth of four kinds of intestinal tract pathogenic bacteria,especially Shigella flexneri with the inhibition zone reaching to(28. 31 ± 0. 27) mm. The results suggested that CICC6238 has high-yield of GABA and potential probiotic properties,which showed good potential for application in functional foods and health-related products.
Lactobacillus plantarum CICC 6238 was screened out for γ-aminobutyric acid( GABA) production from 38 lactic acid bacteria deposited in the China Center of Industrial Culture Collection( CICC). The high-yield GABA strains were obtained by ultraviolet mutagenesis,and its production of GABA was(1. 196 ± 0. 033) g/L,which was 2. 33 times as much as original strain. CICC 6238 was studied extensively for its high tolerance to simulated gastric,intestine juices and bile salts with commercial probiotic strains( Lactobacillus acidophilus NCFM and Lactobacillus rhamnosus HN001) as positive control. After incubation in bile salt environment,the survival rate of CICC 6238 was(36. 90 ± 1. 63) % significantly higher than NCFM [( 15. 16 ± 1. 73) %]. After digestion in artificial gastric juice for 3 hours,the survival rate of CICC6238 was(84. 05 ± 1. 87) %,which was remarkably greater than that of HN001 [(61. 97 ± 2. 45) %]. After incubation in artificial intestine juices for 6 hours,the survival rate of CICC6238 was(61. 50 ± 1. 94) %,which was slightly lower than that of HN001 [(77. 44 ± 3. 08) %]and NCFM [(79.42 ± 2. 08) %]. And CICC 6238 also could inhibit the growth of four kinds of intestinal tract pathogenic bacteria,especially Shigella flexneri with the inhibition zone reaching to(28. 31 ± 0. 27) mm. The results suggested that CICC6238 has high-yield of GABA and potential probiotic properties,which showed good potential for application in functional foods and health-related products.
引文
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[25]张竺英,刘昭明,黄翠姬,等.豆豉中产γ-氨基丁酸乳酸菌的分离鉴定[J].中国调味品,2014(5):53-58.
[26]PARK S Y,LEE J W,LIM S D.The probiotic characteristics and GABA production of Lactobacillus plantarum,K154 isolated from kimchi[J].Food Science&Biotechnology,2014,23(6):1 951-1 957.
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[28]CHO S Y,MIN J P,KIM K M,et al.Production of highγ-aminobutyric acid(GABA)sour kimchi using lactic acid bacteria isolated from mukeunjee,kimchi[J].Food Science&Biotechnology,2011,20(2):403-408.
[2]HAN Yao,LI Hong,LANG Yi-ran,et al.The effects of acute GABA treatment on the functional connectivity and network topology of cortical cultures.[J].Neurochemical Research,2017,42(5):1 394-1 402.
[3]谢明勇,熊涛,关倩倩.益生菌发酵果蔬关键技术研究进展[J].中国食品学报,2014,14(10):1-9.
[4]GUO Zhuang,WANG Ji-cheng,YAN Li-ya,et al.In vitro comparison of probiotic properties of Lactobacillus casei Zhang,a potential new probiotic,with selected probiotic strains[J].LWT-Food Science and Technology,2009,42(10):1 640-1 646.
[5]龚金炎,谢湉,楼坚,等.黄酒浸米液中产γ-氨基丁酸乳酸菌的筛选和鉴定[J].工业微生物,2015,45(6):26-31.
[6]中国食品发酵工业研究院.彩色纳豆生产用菌种及生产工艺:中国,CN201210067238.7[P].2012-10-3.
[7]裴疆森.曲酸生产菌种的筛选和发酵培养条件的优化[J].食品与发酵工业,1997,33(1):11-14.
[8]KWON S Y,GARCIA C V,SONG Y C,et al.GABA-enriched water dropwort produced by co-fermentation with Leuconostoc mesenteroides,SM and Lactobacillus plantarum,K154[J].LWT-Food Science and Technology,2016,73:233-238.
[9]SHEKH S L,DAVE J M,VYAS B R M.Characterization of Lactobacillus plantarum strains for functionality,safety andγ-amino butyric acid production[J].LWT-Food Science and Technology,2016,74:234-241.
[10]LI Wen,WEI Ming-ming,WU Jun-jun,et al.Novel fermented chickpea milk with enhanced level ofγ-aminobutyric acid and neuroprotective effect on PC12 cells[J].Peerj,2016,4(4):e2292.
[11]刘文丽,刘洪霞,张传香,等.泡菜中1株高产γ-氨基丁酸且具有抗菌性乳酸菌株的筛选和鉴定[J].食品与发酵工业,2015,41(12):120-125.
[12]曾林,刘波,许小艳,等.四川泡菜中产γ-氨基丁酸微生物的系统发育与表达能力评估[J].食品科学,2017,38(2):87-91.
[13]DEL P M,MORELLI L,STROZZI G P,et al.Probiotics:from research to consumer[J].Digestive&Liver Disease Official Journal of the Italian Society of Gastroenterology&the Italian Association for the Study of the Liver,2006,38(Suppl 2):S248.
[14]王记成,郭壮,闫丽雅,等.益生菌Lactobacillus caseiZhang与商业益生菌对胃肠转运耐受性及发酵特性的比较[J].中国食品学报,2009,9(5):14-23.
[15]DAMAYANTI E,HERDIAN H,ANGWAR M,et al.Lactic acid bacterial screening from gastrointestinal digestive tract of native and broiler chicken for probiotic candidate purposes[J].Journal of the Indonesian Tropical Animal Agriculture,2012,37(3):168-175.
[16]熊涛,宋苏华,黄锦卿,等.植物乳杆菌NCU116在模拟人体消化环境中的耐受力[J].食品科学,2011,32(11):114-117.
[17]其木格苏都,王记成,孟和毕力格,等.益生菌Lactobacillus casei Zhang和Bifidobacterium lactis V9接种比例及乳固形物含量对其活菌数的影响[J].中国乳品工业,2012,40(1):11-13.
[18]刘彪,张和平.Bifidobacterium lactis V9在脱脂乳粉中贮藏稳定性的研究[J].中国乳品工业,2012,40(12):4-7.
[19]张和平,陈永福.一种瑞士乳杆菌H9及其在制备发酵乳中的用途,CN102329763A[P].2012.
[20]WANG Yu-zhen,LI Yun3xu,XIE Ji-ming,et al.Protective effects of probiotic Lactobacillus casei Zhang against endotoxin-and d-galactosamine-induced liver injury in rats via anti-oxidative and anti-inflammatory capacities[J].International Immunopharmacology,2013,15(1):30-37.
[21]陈世贤,高鹏飞,王记成,等.益生菌Lactobacillus casei Zhang和Bifidobacterium lactis V9在活性乳酸菌饮料中的应用[J].食品与发酵工业,2015,41(11):101-105.
[22]LI Chuan,NIE Shao-ping,ZHU Ke-xue,et al.Effect of Lactobacillus plantarum NCU116 on loperamide-induced constipation in mice[J].International Journal of Food Sciences&Nutrition,2015,66(5):533-538.
[23]熊涛,马晓娟.植物乳杆菌NCU166发酵胡萝卜浆风味物质的分析[J].食品科学,2013,34(2):152-154.
[24]谢俊华,聂少平,丁翘,等.植物乳杆菌NCU116发酵胡萝卜浆对免疫抑制小鼠肠道黏膜免疫的影响[J].食品科学,2015,36(21):201-206.
[25]张竺英,刘昭明,黄翠姬,等.豆豉中产γ-氨基丁酸乳酸菌的分离鉴定[J].中国调味品,2014(5):53-58.
[26]PARK S Y,LEE J W,LIM S D.The probiotic characteristics and GABA production of Lactobacillus plantarum,K154 isolated from kimchi[J].Food Science&Biotechnology,2014,23(6):1 951-1 957.
[27]曾小群,盛姣铃,潘道东,等.新疆酸马奶中产γ-氨基丁酸乳酸菌的筛选与鉴定[J].中国食品学报,2013,13(10):191-196.
[28]CHO S Y,MIN J P,KIM K M,et al.Production of highγ-aminobutyric acid(GABA)sour kimchi using lactic acid bacteria isolated from mukeunjee,kimchi[J].Food Science&Biotechnology,2011,20(2):403-408.