Ha Van Huan, Ho Van Giang, Yong Wang, Siliang Zhang (2011). Overexpression and the enzymatic properties of the recombinant 4 coumarate:coenzyme A ligase, a key enzyme in lignin biosynthesis pathway. J Indian Acad Wood Science (Springer publisher). 8(1):21–25


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Abstract: The investigation for the overexpression and the enzymatic properties of the Pm4CL1 of Pinus mas- soniana Lamb were performed for the first time. The Pm4CL1 cDNA was overexpressed in Escherichia coli Rosetta-gami TM cells. The recombinant Pm4CL1 protein expression was most effective when it was induced with 1 mM isopropyl a-D-thiogalactopyranoside for 2 h at 30C. The 69 His-tagged recombinant Pm4CL1 protein about 60 kDa was purified by agarose coupled with Ni2?-NTA affinity chromatography. The optimal pH and temperature of the reaction were 7.6 and 35C, respectively. The Km and VMax values of the purified Pm4CL1 enzyme for 4-coumaric acid were calculated to be 64.35 lM and 30.92 lM min-1 lg-1, respectively.

Introduction