Screening and identification of strains for producing γ- aminobutyric acid
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摘要: 为获得产γ-氨基丁酸安全菌株,本实验从泡菜汁及酸奶液中进行定向筛选,利用薄层层析法（TLC）对其发酵物进行了初步分析,随后对其进行了16S r DNA基因测序,最后利用高效液相色谱法（HPLC）进行产物定量分析评估其生产能力。结果表明得到六株产谷氨酸脱羧酶催化L-谷氨酸脱羧生成γ-氨基丁酸的菌株,在Gen Bank中进行同源性比对后,鉴定六菌株均为植物乳酸短杆菌。定量分析六株菌发酵液中γ-氨基丁酸含量发现其产量均在200500μg/m L之间,其中最高产量为493.3μg/m L。实验结果显示,硅胶薄层层析可以快速定性检测培养物中是否含有γ-氨基丁酸,而优化后的高效液相色谱法能准确定量检测培养物中的γ-氨基丁酸含量。筛选获得的乳酸杆菌生物安全性高,有一定应用推广前景。Abstract: In order to obtain the safe strains for producing γ- aminobutyric acid,the directed screening for those strains were selected from pickled cabbage juice and yogurt. The preliminary analysis of the fermentation liquor was processed by thin layer chromatography( TLC),and then the 16 S r DNA gene sequencing and Gen Bank comparing were carried for the taxonomy identification. Finally,the high performance liquid chromatography( HPLC) was used for quantitative analysis of γ-aminobutyric acid productivity.Six γ-aminobutyric acid producing strains were screened from the nature samples.The 16 S r DNA gene sequencing and Gen Bank comparing results showed the six strains belonged to short lactic acid bacteria. The productive yield of strains were among 200 ~500 μg / m L,while the maximum yield was 493.3 μg / m L. TLC method can rapidly qualitative detect whether theγ-aminobutyric acid were contained in cultures,and the optimized HPLC method can accurately and quantitatively detect the contents of γ- aminobutyric acid in cultures. Since the biological safety of those lactic acid bacteria,those strains have potential application.
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Keywords:
- glutamate decarboxylase /
- γ-aminobutyric acid /
- screening /
- identification
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