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Expression of dha Operon Required for 1,3-PD Formation in Escherichia coli and Saccharomyces cerevisiae
Authors:Zheng Ma  Zhiming Rao  Liyu Xu  Xiangru Liao  Huiying Fang  Bin Zhuge  Jian Zhuge
Affiliation:(1) The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Research Center of Industrial Microbiology, Jiangnan University, Wuxi, Jiangsu Province, 214122, People’s Republic of China;(2) Applied Science Department, Zhejiang Economic & Trade Polytechnic, 310018 Hangzhou, China;
Abstract:The 1,3-propanediol (1,3-PD) synthesis operon (dha operon) was mainly composed of four genes: dhaB, dhaT, gdrA, and gdrB, which encoded glycerol dehydratase, 1,3-PD oxidoreductase and reactivating factor for glycerol dehydratase, respectively. In the present study, dha operon was cloned from 1,3-PD producing strain Klebsiella pneumoniae. Heterologous expression of cloned dha operon was carried out in Escherichia coli and Saccharomyces cerevisiae W303-1A, respectively. The results indicated that recombinant E. coli harboring the dha operon can produce 8–9 g/l 1,3-PD from glycerol while the 1,3-PD yield of recombinant strain W303-1A-dha could not be detected. In order to complete the 1,3-PD production from glucose, further, we also constructed the recombinant S. cerevisiae W303-1A-BT harboring plasmid pZ-BT. The 1,3-PD production and enzymatic activities of DhaB and DhaT were found in the engineered strain W303-1A-BT. Our results demonstrated that the recombinant S. cerevisiae strain W303-1A-BT that can produce 1,3-PD at low cost was constructed. This study might open a novel way to a safe and cost-efficient method for microbial production of 1,3-PD.
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