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Site-mutation of AroG Gene and Co-expression with TrpBA Gene in Escherichia coli(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2013 05
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Site-mutation of AroG Gene and Co-expression with TrpBA Gene in Escherichia coli
HAO Dali ZHUGE Bin FANG Huiying ZHANG Cheng ZONG Hong ZHUGE Jian
(1Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China) (2Research Center of Industrial Microorganisms, School of Biotechnology, Jiangnan University, Wuxi 214122, China)
tryptophan Escherichia coli aroG trpBA site-directed mutation
Q786 : Q936

TrpBA-encoded tryptophan synthetase (TSase) and aroG-encoded 3-deoxy-D-arabino-heptulosonate-7- phosphate synthase (DAHPS) are key enzymes in tryptophan pathway in Escherichia coli. In order to improve bio-production of tryptophan through bioengineering means, a feedback inhibition resistant aroGfbr (C632T) gene was cloned by SOE-PCR and co-expressed with trpBA gene. The recombinant plasmids pEtac-aroGfbr-tac-trpBA and pEtac-aroG-tac-trpBA were constructed successfully in the recombinant strains, with the enzyme activity analysis indicating an increase of specific activities of DAHPS by 4.3 folds and 3.8 folds, respectively. The enzyme activity of DAHPS after mutation was increased by 1.13 folds. The yield of tryptophan biosynthesis in recombinant strain E. coli JM109/pEtac-aroGfbr-tac-trpBA was increased by 1.4 folds compared with that of the host strains E. coli JM109/pEtac-aroG-tac-trpBA. Fig 5, Tab 2, Ref 15


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Last Update: 2013-10-28