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Citation:

Enzymatic Characteristics of 4-coumarate: coenzyme A Ligase (4CL3) from Populus tomentosa

  • Received Date: 2012-03-19
  • A new 4CL gene from Populus tomentosa Carr. was cloned and named as 4CL3. Bioinformatics method was used to analysis the protein sequence of this 4CL3 gene aligning with two other 4CL proteins. The authors found the conserved sequence Box I and Box II exist in the 4CL gene family. Prokaryotic expression way was used to research the enzyme function of this 4CL3 protein, the result showed that the 4CL protein had different enzymatic activity towards the different substrate. The 4CL3 protein had the biggest enzymatic activity at the temperature of 40 ℃, and it had the biggest enzymatic activity at pH 8.5, this enzyme has the ability of resisting alkaline environment. The 5 substrates of 4CL were mixed and the method of HPLC-MS was used to survey the consumption of the substrate after putting 4CL enzyme to start the reaction. It was found that besides sinapic there was no consumption of caffeic either.
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Enzymatic Characteristics of 4-coumarate: coenzyme A Ligase (4CL3) from Populus tomentosa

  • 1. Key Laboratory of Tree Breeding and Cultivation, State Forestry Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China

Abstract: A new 4CL gene from Populus tomentosa Carr. was cloned and named as 4CL3. Bioinformatics method was used to analysis the protein sequence of this 4CL3 gene aligning with two other 4CL proteins. The authors found the conserved sequence Box I and Box II exist in the 4CL gene family. Prokaryotic expression way was used to research the enzyme function of this 4CL3 protein, the result showed that the 4CL protein had different enzymatic activity towards the different substrate. The 4CL3 protein had the biggest enzymatic activity at the temperature of 40 ℃, and it had the biggest enzymatic activity at pH 8.5, this enzyme has the ability of resisting alkaline environment. The 5 substrates of 4CL were mixed and the method of HPLC-MS was used to survey the consumption of the substrate after putting 4CL enzyme to start the reaction. It was found that besides sinapic there was no consumption of caffeic either.

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