Identification and utilization of a 1,3-propanediol oxidoreductase isoenzyme for production of 1,3-propanediol from glycerol in Klebsiella pneumoniae

• Published in: Applied microbiology and biotechnology Vol. 85; no. 3; pp. 659 - 666
• Main Authors: Seo, Jeong-Woo, Seo, Mi-Young, Oh, Baek-Rock, Heo, Sun-Yeon, Baek, Jin-Oh, Rairakhwada, Dina, Luo, Lian Hua, Hong, Won-Kyung, Kim, Chul Ho
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 2010; Springer-Verlag; Springer; Springer Nature B.V
• Subjects: Alcohol Dehydrogenase; Alcohol Oxidoreductases - genetics; Alcohol Oxidoreductases - metabolism; Amino acids; Applied Genetics and Molecular Biotechnology; Bacteria; Biological and medical sciences; Biomedical and Life Sciences; Biosynthetic Pathways - genetics; Biotechnology; By products; Dehydrogenases; E coli; Enzymes; Escherichia coli; Escherichia coli - genetics; Fermentation; Fundamental and applied biological sciences. Psychology; Gene Deletion; Gene Dosage; Gene Expression; Genetic Complementation Test; Glycerol; Glycerol - metabolism; Isoenzymes - genetics; Isoenzymes - metabolism; Kinases; Klebsiella pneumoniae; Klebsiella pneumoniae - genetics; Klebsiella pneumoniae - metabolism; Life Sciences; Metabolism; Microbial Genetics and Genomics; Microbiology; Mutation; Plasmids; Pneumonia; Polymerization; Propylene Glycols - metabolism; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Studies; Textile fibers; 1,3-Propanediol; 1,3-Propanediol oxidoreductase isoenzyme; Glycerol metabolism; Enzyme; Isozyme; Glycerol; Identification; Metabolism; Production; Bacteria; Oxidoreductases; Propanediol; Klebsiella pneumoniae; Enterobacteriaceae
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-009-2123-4

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.