Microbial transformation of benzene to cis-3,5-cyclohexadien-1,2-diols by recombinant bacteria harboring toluene dioxygenase gene tod.
Toluene dioxygenase (TDO) catalyzes asymmetric cis-dihydroxylation of aromatic compounds. To achieve high efficient biotransformation of benzene to benzene cis-diols, Pseudomonas putida KT2442, Pseudomonas stutzeri 1317, and Aeromonas hydrophila 4AK4 were used as hosts to express TDO gene tod. Plasmid pSPM01, a derivative of broad-host plasmid pBBR1MCS-2 harboring tod from plasmid pKST11, was constructed and introduced into the above three strains. Their abilities to catalyze the biotransformation of benzene to benzene cis-diols, namely, cis-3,5-cyclohexadien-1,2-diols abbreviated as DHCD, were examined. In shake-flask cultivation under optimized culture media and growth condition, benzene cis-diols production by recombinant P. putida KT2442 (pSPM01), P. stutzeri 1317 (pSPM01), and A. hydrophila 4AK4 (pSPM01) were 2.68, 2.13, and 1.17 g/l, respectively. In comparison, Escherichia coli JM109 (pSPM01) and E. coli JM109 (pKST11) produced 0.45 and 0.53 g/l of DHCD, respectively. When biotransformation was run in a 6-l fermenter, DHCD production in P. putida KT2442 (pSPM01) was approximately 60 g/l; this is the highest DHCD production yield reported so far.
Ouyang
Shao-Ping
SP
Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, 100084, China.
Sun
Shang-Yu
SY
Liu
Qian
Q
Chen
Jinchun
J
Chen
Guo-Qiang
GQ
D016980:Aeromonas hydrophila / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D001554:Benzene / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D003511:Cyclohexanols / Q000378:metabolism*; D005285:Fermentation; D005818:Genetic Engineering / Q000379:methods; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D044223:Pseudomonas stutzeri / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D003511:Cyclohexanols; D011994:Recombinant Proteins; D010105:Oxygenases; C023172:toluene dioxygenase; D001554:Benzene