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高产γ-氨基丁酸的棉子糖肠球菌的筛选、鉴定及其摇瓶发酵条件的优化
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国家863计划项目(No. 2012AA021302); 国家973计划项目(No. 2011CB707401, 2013CB734004); 天津市滨海新区自主创新重大项目(No. 2011-BK120014)


Screening, identification and flask fermentation optimization of a high-yield γ-aminobutyric acid Enterococcus raffinosus strain
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    摘要:

    【目的】在中国传统发酵泡菜中分离出高产?γ-氨基丁酸(γ-aminobutyric acid, GABA)的乳酸菌。【方法】对分离自泡菜中的菌株M1进行形态观察、生理生化特性鉴定及其16S rDNA序列分析, 实验采用单因素和正交设计对以MRS培养基为基础的GABA发酵培养基与摇瓶发酵条件进行了优化。【结果】菌株M1的形态培养和生理生化特征均符合肠球菌属(Enterococcus)特征, 其16S rDNA序列与Enterococcus raffinosus SS1278 16S rDNA序列同源性达 99%, 鉴定为棉子糖肠球菌(Enterococcus raffinosus)。优化该菌株产GABA的发酵培养基的实验发现, 最佳摇瓶发酵条件为: 接种量10%, 发酵温度30 °C, 培养初始pH 5.5, 发酵周期60 h, 谷氨酸一钠底物浓度 10%, GABA产量提高了1.22 倍。【结论】棉子糖肠球菌(E. raffinosus) M1菌株具有工业化发酵生产GABA的潜力。

    Abstract:

    [Objective] A high-yield γ-aminobutyric acid (GABA) producing lactic acid bacterium (LAB), strain M1 was isolated from pickled Chinese vegetables by our laboratory. [Methods] Its physiological and biochemical characteristics and 16S rDNA sequence were analyzed. GABA fermentation medium, based on MRS medium, was optimized using single factor test and orthogonal design, as well as the shake flask condition for GABA fermentation. [Results] The results indicated that the morphological, physiological and biochemical characteristics of strain M1 were accorded with Enterococcus family. The identity between the obtained 16S rDNA sequence of strain M1 and Enterococcus raffinosus SS1278 was up to 99%, strain M1 was accordingly identified as Enterococcus raffinosus. Moreover, its GABA fermentation medium was optimized, and the shake flask condition for GABA fermentation was: inoculum of 10%, growing temperature at 30 °C, initial pH at 5.5, fermentation period of 60 h and monosodium glutamate substrate concentration of 10%. Under these conditions, GABA yield by flask fermentation achieved a 1.22-fold increase. [Conclusion] The isolated E. raffinosus M1 strain reveals its potential for industrial fermentation of GABA.

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陈慧敏,高强,苏喆,王梦晓,段强,张云泽,司晓光. 高产γ-氨基丁酸的棉子糖肠球菌的筛选、鉴定及其摇瓶发酵条件的优化[J]. 微生物学通报, 2012, 39(11): 1642-1652

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  • 在线发布日期: 2012-11-21
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