产γ-氨基丁酸益生菌的筛选及其生物活性研究
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- 英文论文题名：Screening of γ-Aminobutyric acid-producing probiotics and characterization of its biological activity
- 论文作者：徐毓琴 ; 戴茜茜 ; 唐慧琴 ; 谷笑笑 ; 潘康成
- 英文论文作者：XU Yu-qin ; DAI Xi-xi ; TANG Hui-qin ; GU Xiao-xiao ; PAN Kang-cheng ; College of Veterinary Medicine ; Sichuan Agricultural University ; Key Laboratory of Animal Disease and Human Health of Sichuan Province
- 年：2016
- 作者机构：四川农业大学动物医学院;动物疫病与人类健康四川省重点实验室;
- 论文关键词：益生菌 ; γ-氨基丁酸 ; 生物拮抗 ; 耐酸耐胆盐
- 英文论文关键词：probiotics ; γ-aminobutyric acid ; bioantagonism ; acid and bile salt tolerance
- 会议召开时间：2016-11-04
- 会议录名称：中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会论文集
- 语种：中文
- 分类号：S852.6
- 学会代码：ZGXJ
- 会议名称：中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会
- 会议地点：中国山东青岛
- 主办单位：中国畜牧兽医学会动物微生态学分会
- 学会名称：中国畜牧兽医学会
- 页数：1
- 文件大小：81k
- 原文格式：O
- 会议级别：全国
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序列分析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;该菌株菌悬液对鸡白痢沙门氏菌、大肠杆菌和金黄色葡萄球菌有一定的抑制作用,其上清液无抑菌能力;该菌初始活菌数为10~8cfu·mL~(-1)时,在pH 2.0的酸性环境中处理4 h后,活菌数能达到10~6 cfu·mL~(-1)以上;0.3%胆盐处理4 h,活菌数达到10~3 cfu·mL~(-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 with 16 S rDNA 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.GA8 strain had some inhibitory effect on Salmonella pullorum,Escherichia coli and Staphylococcus aureus,and the supernatant of GA8 strain culture had no bacteriostasis ability.When the initial concentration of living bacteria was 10~8 cfu·mL~(-1),the living bacteria could reach more than 10~6 cfu·mL~(-1) after cultured on medium with pH 2.0 for 4 h and10~3 cfu·mL~(-1) after cultured on medium contained bile salt 0.3%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.
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 with 16 S rDNA 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.GA8 strain had some inhibitory effect on Salmonella pullorum,Escherichia coli and Staphylococcus aureus,and the supernatant of GA8 strain culture had no bacteriostasis ability.When the initial concentration of living bacteria was 10~8 cfu·mL~(-1),the living bacteria could reach more than 10~6 cfu·mL~(-1) after cultured on medium with pH 2.0 for 4 h and10~3 cfu·mL~(-1) after cultured on medium contained bile salt 0.3%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.
引文
[1]ROBERRTS E,FRANKEL S.γ-Aminobutyric acid in brain:Its formation from glutamic acid[J].Journal of Biological Chemistry,1950,187(1):55-63.
[2]ALI B H,AL-SALAM S,ZA'ABI M A,et al.Renoprotective effects of gamma-Aminobutyric acid on cisplatin-induced acute renal injury in rats[J].Basic&Clinical Pharmacology&Toxicology,2014,116(1):62-68.
[3]贤乾隆.产γ-氨基丁酸乳酸菌的筛选及其功能性发酵酸奶的研制[D].柳州:广西科技大学,2013
[2]ALI B H,AL-SALAM S,ZA'ABI M A,et al.Renoprotective effects of gamma-Aminobutyric acid on cisplatin-induced acute renal injury in rats[J].Basic&Clinical Pharmacology&Toxicology,2014,116(1):62-68.
[3]贤乾隆.产γ-氨基丁酸乳酸菌的筛选及其功能性发酵酸奶的研制[D].柳州:广西科技大学,2013