摘要
对荧光假单胞菌合成2,4-二乙酰基间苯三酚(2,4-DAPG)操纵子的关键基因phlD进行了克隆,并采用E.coliBL21(DE3)/pET28 a(+)对phlD进行了诱导表达,其指导合成的PhlD蛋白融合表达后为42000 Da.生物信息学方法分析PhlD具有PKSⅢ型聚酮合酶的结构及催化聚酮缩合、酰基转移反应的功能,并推测PhlD催化底物丙二酸单酰辅酶A经过聚酮缩合途径合成间苯三酚及其衍生物.对重组菌进行了摇瓶发酵并检测出发酵液中确有产物间苯三酚的合成.
The critical gene phlD for 2, 4-DAPG biosynthesis from Pseudornonas fluorescens was cloned and expressed in E. coli BL21 （DE3）/pET28a（ ＋ ）/phlD recombinant strain. The gene of phlD was amplified using Pseudornonas fluorescens 2P24 genome as template. The PhlD of 42 000 Da fusion protein was induced and expressed in E. coli BL21（DE3）/pET28a（＋） system. The phlD gene sequence was analyzed, the senior structure of the protein coded by the phlD also forecasted. PhlD was one of polyketide synthase of type-Ⅲ, and the function of PhlD was similar to the polyketal synthase and acyltransferase. The fermentation liquid of recombinant strain was detected, and synthesis of the phloroglucinol was implemented.
出处
《北京理工大学学报》
EI
CAS
CSCD
北大核心
2009年第5期465-470,共6页
Transactions of Beijing Institute of Technology
基金
国家部委基金资助项目(62301110604)
