Expression of dha operon required for 1,3-PD formation in Escherichia coli and Saccharomyces cerevisiae.
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.
Ma
Zheng
Z
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.
Rao
Zhiming
Z
Xu
Liyu
L
Liao
Xiangru
X
Fang
Huiying
H
Zhuge
Bin
B
Zhuge
Jian
J
D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose / Q000378:metabolism; D009876:Operon / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D004271:DNA, Fungal; D005656:Fungal Proteins; D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; C052370:1,3-propanediol dehydrogenase; D005947:Glucose