产谷氨酸脱羧酶金银花内生菌的鉴定及其转化特性研究

食品科学 ›› 2007, Vol. 28 ›› Issue (2): 196-202.

产谷氨酸脱羧酶金银花内生菌的鉴定及其转化特性研究

杨胜远, 陆兆新, 孙力军, 别小妹, 吕凤霞, 曾晓雄

南京农业大学食品科技学院；南京农业大学食品科技学院江苏南京210095.广西大学生命科学与技术学院；广西南宁530005；江苏南京210095；

出版日期:2007-02-15 发布日期:2011-12-31

Study on Identification of Endophyte with Glutamate Decarboxylase Activity from Honeysuckle and Its Biotransformation Properties

YANG Sheng-Yuan, LU Zhao-Xin, SUN Li-Jun, BIE Xiao-Mei, 吕Feng-Xia , ZENG Xiao-Xiong

1.College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China; 2.College of Life Science and Technology, Guangxi University, Nanning 530005, China

Online:2007-02-15 Published:2011-12-31

摘要/Abstract

摘要： 从金银花植株茎中分离到了1株具有谷氨酸脱羧酶活性的内生细菌,1g菌体(DW)24h可将241.224μmolL-Glu转化生成GABA。通过引物设计,利用PCR扩增出该菌株的16SrDNA序列,大小为1459bp。通过形态特征、生理生化特征和16SrDNA序列分析鉴定EJH-7为巨大芽孢杆菌(Bacillus megaterium)。同时基于16SrDNA构建了系统进化树,并对EJH-7进行了系统发育分析。Bacillus megateriumEJH-7细胞转化谷氨酸生成GABA的最适反应温度和pH分别为30℃和5.5;表面活性剂Tween20和Tween80对转化反应活性有抑制作用,而Triton100影响不显著;Ca2+、Cu2+和Co2+对转化反应活性有促进作用,分别提高了20.36%、46.61%和6.77%,而K+、Fe2+、Zn2+、Mg2+和Mn2+有不同程度的抑制作用。

关键词: 谷氨酸脱羧酶, 金银花, 植物内生菌, 巨大芽孢杆菌, 16S rDNA, 生物转化

Abstract: An endophytic bacterium strain EJH-7 with glutamate decarboxylase activity, transforming 241.224μmol L- glutamate to GABA in 24h by 1g cell (DW), was isolated and screened from the stem of Lonicera japonica Thunb. Through the morphologic, physiological and biochemical characteritics and the 16S rDNA sequence analysis, EJH-7 has been identified as Bacillus megaterium. Also, the phylogenesis was analyzed through the phylogenetic tree base on the 16S rDNA sequences, indicating that strain EJH-7 belonged to Bacteria, Firmicutes, Bacillales, Bacillaceae, Bacullus. The properties of the biotrans- formation (converting L-glutamate to GABA) of Bacillus megaterium EJH-7 were also examined. As a result, the optimum temperature and pH value for biotransformation are 30℃ and 5.5 respectively. The biotransformation activities are inhibited by ..Tween 20 and Tween 80, and hardly affected by Triton 100. The biotransformation activities of Bacillus megaterium EJH-7 are significantly increased by 2.5mmol/L of Ca2+, Cu2+ and Co2+, and raised by 20.36%, 46.61% and 6.77% respectivelyin different degree by K+, Fe2+, Zn2+, Mg2+ and Mn2+.

Key words: glutamate decarboxylase, honeysuckle, endophyte, Bacillus megaterium, 16s rDNA, biotransformation

杨胜远, 陆兆新, 孙力军, 别小妹, 吕凤霞, 曾晓雄. 产谷氨酸脱羧酶金银花内生菌的鉴定及其转化特性研究[J]. 食品科学, 2007, 28(2): 196-202.

YANG Sheng-Yuan, LU Zhao-Xin, SUN Li-Jun, BIE Xiao-Mei, 吕Feng-Xia , ZENG Xiao-Xiong. Study on Identification of Endophyte with Glutamate Decarboxylase Activity from Honeysuckle and Its Biotransformation Properties[J]. FOOD SCIENCE, 2007, 28(2): 196-202.

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