产γ-氨基丁酸益生菌的筛选及其生物活性研究  

Screening of γ-aminobutyric acid-producing probiotics and characterization of its biological activity

作　　者：徐毓琴 戴茜茜[1] 唐慧琴[1] 谷笑笑 潘康成[1,2] 

机构地区：[1]四川农业大学动物医学院,四川成都611130 [2]动物疫病与人类健康四川省重点实验室,四川成都611130

出　　处：《浙江农业学报》2016年第3期502-508,共7页Acta Agriculturae Zhejiangensis

基　　金："教育部长江学者和创新团队发展计划"创新团队(IRT0848);四川农业大学教学改革研究项目(X2011008)

摘　　要：为筛选具有生物拮抗活性和耐酸耐胆盐能力的高产γ-氨基丁酸(GABA)的菌株,首先从待分离样品中筛选出可疑菌株,然后利用纸层析和HPLC-MS进行定性和定量分析,筛选出高产γ-氨基丁酸菌株。通过细菌形态、生理生化特性并结合细菌16S rDNA序列对高产菌株进行鉴定,同时研究高产菌株对3株动物性病原菌的生物拮抗作用及其耐酸耐胆盐试验。结果表明,从泡菜中筛选出的菌株GA8为高产γ-氨基丁酸菌株,在含有1%L-谷氨酸钠的GYP培养基中37℃培养48 h,产GABA量达3.50 g·L^(-1),经鉴定该菌株为植物乳杆菌;该菌株全菌液和上清液对鸡白痢沙门氏菌、鸡源大肠杆菌和金黄色葡萄球菌有一定的抑制作用,其菌体悬液无抑菌能力;该菌初始活菌数为108cfu·m L^(-1)时,在pH 2.0的酸性环境中处理4 h后,活菌数能达到106cfu·m L^(-1)以上;0.3%胆盐处理4 h,活菌数达到10~3cfu·m L^(-1)左右。表明所筛选出的产γ-氨基丁酸菌株GA8具有一定的抑菌性能和耐酸能力,但耐胆盐能力不强。The purpose of the present study was to screen a high γ-aminobutyric acid（ GABA）-producing probiotics,which had the ability of biological antagonism and tolerance of acid and bile salt. The suspicious strains which producing GABA were analyzed qualitatively and quantitively by paper chromatography and high performance liquid chromatography and mass spectrometry（ HPLC-MS）. Combining the morphological,physiochemical properties with16 S r DNA sequencing analysis,a high GABA-producing bacterium was identified. Then the inhibition of this high GABA-producing strain against three pathogen bacteria from animal and the tolerance of acid and bile salt were performed. The results showed that the strain GA8 which isolated from pickle had high yield of GABA; it was cultured in GYP medium containing 1% L-glutamate at 37 ℃ for 48 h,the yield of GABA reached 3. 50 g·L^（–1）; it was identified as Lactobacillus plantarum. The bacteria liquid and supernatant of GA8 strain culture had some inhibitory effect on Salmonella pullorum,Escherichia coli and Staphylococcus aureus,and GA8 strain had no bacteriostasis ability.When the initial concentration of living bacteria was 108cfu·m L^（–1）,the living bacteria could reach more than 106cfu·m L^（–1）after cultured on medium with p H 2. 0 for 4 h and 103cfu·m L^（–1）after cultured on medium contained bile salt0. 3% for 4 h. The results indicated that the strain GA8 had some antibacterial properties and acid resistance,but bile salt resistant ability was not prominent.

关 键 词：益生菌 Γ-氨基丁酸 生物拮抗 耐酸耐胆盐 

分 类 号：S667.4[农业科学—果树学]