收稿日期: 2011-03-14
修回日期: 2011-04-04
网络出版日期: 2011-08-02
基金资助
国家"863”计划重点项目( 2007AA100402)
Yeast Surface Display of LAI from Lactobacilllus plantarum
Received date: 2011-03-14
Revised date: 2011-04-04
Online published: 2011-08-02
Supported by
国家"863”计划重点项目( 2007AA100402)
刘佩 阮晖 沈生荣 周倩 马鎏鏐 何国庆 . 植物乳杆菌LAI 的酵母表面展示[J]. 华南理工大学学报(自然科学版), 2011 , 39(9) : 140 -146 . DOI: 10.3969/j.issn.1000-565X.2011.09.024
In order to investigate the function of linoleic acid isomerase( LAI) and construct the recombinant yeast strains,first,the lai gene was cloned from Lactobacillus plantarum lp15-2-1,with its coding sequence being also analyzed. Then,lai was fused to the sequences of both the signal peptide of yeast and the anchor region of α-agglutinin,thus constructing the yeast cell-surface display vector. Finally,the vector was transformed into Saccharomyces cerevisiae K601 by means of the electroporation,and the transformants were screened by using the synthetic dropout medium without uracil. Amino acid sequence analysis shows that LAI is of a high homology to the myosin-cross-reactive antigen proteins as well as of a semi-conserved flavin adenine dinucleotide binding motif near the N-terminus. The activity results of the recombinant yeast strains demonstrate that LAI from Lactobacillus plantarum is successfully displayed on the cell-surface of yeast K601,and that the activity of LAI reaches the maximum 40.5U/mL in 48 h. Moreover,GC results indicate that the recombinant yeast strains are capable of producing a single c9,t11- CLA isomer.
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