产γ-氨基丁酸乳杆菌高效筛选方法的建立
Establishment of high throughput screening method for γ-aminobutyric acid-producing lactobacillus
范城飞 1翟征远 1王然 2郝彦玲 2孙婷 3马霞 3刘福东 3赵雯 3洪维錬3
作者信息
- 1. 中国农业大学食品科学与营养工程学院,北京 100083
- 2. 中国农业大学营养与健康系,北京 100193
- 3. 内蒙古乳业技术研究院有限责任公司,呼和浩特 010110;内蒙古伊利实业集团股份有限公司,呼和浩特 010110
- 折叠
摘要
本研究首先发现11株短乳杆菌皆产生大量气体导致培养管盖子弹起,进一步对48 h发酵液pH进行测定,发现有3株菌的发酵液具有较高pH,薄层层析结果表明产生大量气体且发酵液pH较高的菌株能够产生γ-氨基丁酸(GABA),进一步通过质谱实验验证,产量分别是124.14、203.00、211.50 mg/L.通过高效测序发现11株短乳杆菌GABA合成基因簇结构和核酸序列基本相同,因此在基因层面无法实现高效筛选.根据GABA生物合成过程中产生二氧化碳及消耗氢离子的现象建立高效筛选方法.进一步对220株植物乳杆菌进行产气观察结合pH测定判定菌株是否产GABA,并通过薄层色谱进行验证,表明此方法可行.该方法具有低成本高效的特点,能够快速从大量乳杆菌中筛选出产GABA菌株.
Abstract
Firstly,11 strains of Levilactobacillus brevis were found to produce a large amount of gas,which was enough to push the cover of culture tubes open.Among them,three strains showed a higher pH after 48 h fermentation.Then,thin layer chromatography analysis further proved that only the strains with gas production and higher pH of the fermentation broth could produce gamma-aminobutyric acid(GABA).Mass spectrom-etry showed that the yield of GABA of three L.brevis strains was 124.14,203.00 mg/L and 211.50 mg/L,respectively.Interestingly,high-through-put sequencing analysis found that the gene structure and nucleic acid sequence responsible for GABA synthesis of 11 L.brevis strains were almost same,so it may not be achieved for high-throughput screening GABA-producing strains at the gene level.Based on carbon dioxide production and hydrogen ions consumption during fermentation,a high-throughput method for screening GABA-producing lactobacillus strains was estab-lished.Furthermore,the established method was applied to select GABA-producing Lactiplantibacillus plantarum among 220 bacterial strains,which proved its feasibility.In conclusion,this study reported an available method for quickly screening GABA-producing strains from a large number of lactobacillus with low cost and high throughput characteristics.
关键词
γ-氨基丁酸/乳杆菌/高效Key words
GABA/Lactobacillus/high throughput screening引用本文复制引用
出版年
2024
NETL
NSTL
维普
万方数据