Identification and utilization of a 1,3-propanediol oxidoreductase isoenzyme for production of 1,3-propanediol from glycerol in Klebsiella pneumoniae.
In a previous study, we showed that 1,3-propanediol (1,3-PD) was still produced from glycerol by the Klebsiella pneumoniae mutant strain defective in 1,3-PD oxidoreductase (DhaT), although the production level was lower compared to the parent strain. As a potential candidate for another putative 1,3-PD oxidoreductase, we identified and characterized a homolog of Escherichia coli yqhD (88% homology in amino acid sequence), which encodes an alcohol dehydrogenase and is well known to replace the function of DhaT in E. coli. Introduction of multiple copies of the yqhD homolog restored 1,3-PD production in the mutant K. pneumoniae strain defective in DhaT. In addition, by-product formation was still eliminated in the recombinant strain due to the elimination of the glycerol oxidative pathway. An increase in NADP-dependent 1,3-PD oxidoreductase activity was observed in the recombinant strain harboring multiple copies of the yqhD homolog. The level of 1,3-PD production during batch fermentation in the recombinant strain was comparable to that of the parent strain; further engineering can generate an industrial strain producing 1,3-propanediol.
Seo
Jeong-Woo
JW
Molecular Bioprocess Research Center, Jeonbuk Branch Institute, KRIBB, Jeongeup, Jeonbuk, South Korea.
Seo
Mi-Young
MY
Oh
Baek-Rock
BR
Heo
Sun-Yeon
SY
Baek
Jin-Oh
JO
Rairakhwada
Dina
D
Luo
Lian Hua
LH
Hong
Won-Kyung
WK
Kim
Chul Ho
CH
D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D053898:Biosynthetic Pathways / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D017353:Gene Deletion; D018628:Gene Dosage; D015870:Gene Expression; D005816:Genetic Complementation Test; D005990:Glycerol / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D007711:Klebsiella pneumoniae / Q000235:genetics* / Q000378:metabolism*; D011409:Propylene Glycols / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D007527:Isoenzymes; D011409:Propylene Glycols; C041787:1,3-propanediol; D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; C052370:1,3-propanediol dehydrogenase; D005990:Glycerol