军事医学2012，Vol.36Issue(2)：124-127,4.
产L-苯丙氨酸工程菌的构建及不同碳源对其产量的影响
Construction of L-phenylalanine-producing strain and effect of different carbon sources on yield of L-phenylalanine
黄坤央 1芦佳 2赵越 3徐琪寿 3黄英武2
作者信息
- 1. 郑州大学药学院,郑州,450001
- 2. 军事医学科学院放射与辐射医学研究所,北京,100850
- 3. 内蒙古农业大学食品科学与工程学院,呼和浩特,010018
- 折叠
摘要
Abstract
Objective To construct the L-phenylalanine-producing engineering strains and observe the effect of glucose and glycerol, when used as carbon source, on the growth of recombination strains. Methods GlpK gene was amplified by PCR and changes were made in its ribosome binding site( RBS ) sequence, respectively before both were attached to pZE12-AF vectors. Two recombinant plasmids, pZE12-AF~RBS I -glpK and pZE12-AF-RBS II -glpK, were obtained. Then, they were transformed into Esherichia coli MGA separately for the protein expression by SDS-PAGE. The effect on the yield of L-phenylalanine was assayed by HPLC after fermentation. Results The expression level of glpk gene in the engineering strain of E. Coli MGApZE12-AF- RBS I -glpK was higher than that in E. Coli MG ApZE12-AF-RBS I -glpK strain. When glycerol was used as carbon source, L-phenylalanine in the engineering strain E. Coli MG ApZE12-AFRBS II-glpK was 14. 16 times the production of the engineering strain E. Coli MGApZE12-RBS-AF, 3. 18 times that of the engineering strain E. Coli MGApZE12-AF and 63. 31 times that of the native strain E. Coli MGA. The L-phenylalanine productivity of all the strains was higher when glycerol was used as the carbon source than when glucose was used. Conclusion Changes of RBS sequence and the expression of glpK gene could increase the output of L-phenylalanine in the engineering strain, and glycerol is a better carbon source for recombination strains than glucose.关键词
甘油激酶/核糖体结合位点/L-苯丙氨酸/发酵/聚合酶链反应Key words
glycerol kinase/ ribosome binding site/ L-phenylalanine/ fermentation/ polymerase chain reaction分类
生物科学引用本文复制引用
黄坤央,芦佳,赵越,徐琪寿,黄英武..产L-苯丙氨酸工程菌的构建及不同碳源对其产量的影响[J].军事医学,2012,36(2):124-127,4.
