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| Visualization of Mutation to Structure Homologous Modeling, Site-Directed Mutagenesis and Express D-Lactate Dehydrogenase from Aquifex aeolicus |
| TIAN Jin-hong1,2,3, LIU Qi2, ZHAN Li-ping2, ZHOU Ze-yang1,4 |
1. Institute of Sericulture and Systems Biology,Southwest University,Chongqing 400716,China;
2. College of Pharmaceutical Sciences,Southwest University,Chongqing 400716,China;
3. Chongqing Engineering Technology Research Centre of Veterinary Drugs,Chongqing 400716,China;
4. College of Life Science,Chongqing Normal University,Chongqing 400047,China |
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Abstract Objective to construct high yields recombinant E. coli strains producing phenyl lactic acid(PLA). Methods Analyze the X-ray structure of active site amino acid residues of D-LDH from Aquifex aeolicus, and compare it with mutant D-LDH of the homologous model. The optimized 3D structures of the mutant enzyme were elected by analyzing the space conformation of the amino acid residues in substrate binding domain of the mutant D-LDH. The genes of enzyme were site-directed mutated, cloned and expressed. Results F49A or Y297S mutants were finally elected for single-site mutation models, F49A/Y297S mutant was elected for double -site mutations, and three recombinant E. coli strains were proved for PLA production. Conclusion The method of mutation visualization is an effective method for construction of high yield gene engineering bacterium producing PLA.
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Received: 31 January 2012
Published: 25 May 2012
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