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›› 2009, Vol. 9 ›› Issue (4): 791-795.

• 生化工程专栏 • Previous Articles     Next Articles

Biosynthesis of Poly-g-glutamate Acid by Escherichia coli

MA Jie, WANG Dan, LI Qiang, XING Jian-min, LIU Hui-zhou,   

  1. State Key Laboratory of Multi-Phase＆ Complex Systems, Institute of Process Engineering, Chinese Academy of Sciences Department of Applied Chemistry, Shenyang Institution of Chemical Technology State Key Laboratory of Multi-Phase＆ Complex Systems, Institute of Process Engineering, Chinese Academy of Sciences Lab. Sep. Sci. & Eng., Institute of Process Engineering, Chinese Academy of Sciences Institute of Process Engineering, Chinese Academy of Sciences
  • Received:2009-03-10 Revised:2009-05-07 Online:2009-08-20 Published:2009-08-20
  • Contact: MA Jie,

基因工程大肠杆菌合成g-聚谷氨酸

马婕 王丹 李强 邢建民 刘会洲   

  1. 中国科学院过程工程研究所生化工程国家重点实验室 沈阳化工学院应用化学学院 中国科学院过程工程研究所 中国科学院过程工程研究所分离科学与工程青年实验室 中国科学院过程工程研究所分离工程与工程青年实验室
  • 通讯作者: 马婕

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Abstract: The genes, ywsC, ywtA and ywtB, involving in biosynthesis of poly-γ-glutamic acid were amplified by PCR from the genome of Bacillus subtills 168. Sequence blast result showed that the sequence of target genes was 100% the same as that of B. subtilis 168. The genes were then inserted to the plasmid pTrcHisA and transformed into E. coli TOP10 and E. coli BL21 (DE3) host cells. The result indicated that both the strains obtained the ability to biosynthesize poly-g-glutamic acid, and the highest yield of g-PGA reached 0.134 g/L.

Key words: g-PGA, Bacillus subtilis 168, E. coli TOP10, E. coli BL21 (DE3)

摘要： 利用PCR方法，从自身不合成g-聚谷氨酸(g-PGA)的Bacillus subtilis 168菌的基因组DNA中扩增出g-PGA合成基因ywsC, ywtA和ywtB，测序并对该基因编码区进行序列分析，比对结果表明，扩增的ywsC, ywtA和ywtB与文献报道的相似度为100%. 将3个基因连接到pTrcHisA载体后转化至E. coli TOP10及E. coli BL21 (DE3)宿主菌表达，结果宿主菌细胞均具备了g-PGA生物合成能力，产量最高达到0.134 g/L.

关键词: g-聚谷氨酸, 枯草芽孢杆菌B. subtilis 168, 大肠杆菌TOP10, E. coli BL21 (DE3)

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