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Construction of the Mutants of L.plantarum P-8 Linoleic Acid Isomerase Using PCR Mediated Site-directed Mutagenesis
Author(s): 
Pages: 5-8
Year: Issue:  15
Journal: Farm Products Processing

Keyword:  conjugated linoleic acidlinoleic acid isomeraseessential amino acidssite-directed mutagenesis;
Abstract: In this study linoleic acid isomerase of Lactobacillus plantarum subsp. P-8 as the research object,at online tools and software such as http://swissmodel.expasy.org/, http://smart.embl-heidelberg.de/ and vecter NTI to analysis the bioinformatics of linoleic acid isomerase and predict the site of the enzyme activity related. Predicted results confirme the three necessary amino acid residues of linoleic acid isomerase:the 68 th Glycine,the 107 th Arginine and the 172 th Histidine. And designed a pair of primer containing mutations,recombinant plasmid p QE30-LAI as a template,using PCR mediated site directed mutagenesis constructed mutants determined by sequencing alignment,the 68 th Glycine mutated into Alanine,the107 th Arginine mutated into Leucine,the 172 th Histidine mutated into Proline. Successfully are constructed mutant G68 A,R107L and H172 P that lay a foundation for further study of the LAI’s structure and function.
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