Production of 3-hydroxypropionic acid through propionaldehyde dehydrogenase PduP mediated biosynthetic pathway in Klebsiella pneumoniae

• Published in: Bioresource technology 2012-01, Vol.103 (1), p.1-6
• Main Authors: Luo, Lian Hua, Kim, Chul Ho, Heo, Sun-Yeon, Oh, Baek-Rock, Hong, Won-Kyung, Kim, Seonghun, Kim, Dae-Hyuk, Seo, Jeong-Woo
• Format: Article
• Language: English
• Subjects: 3-hydroxypropionic acid; aldehyde dehydrogenase; Aldehydes - metabolism; Amino Acid Sequence; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; biochemical pathways; Biological and medical sciences; biosynthesis; Biosynthetic Pathways - drug effects; Cell death; cell growth; enzyme activity; Escherichia coli; Escherichia coli - drug effects; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Gene Deletion; Genes; Glycerol; Glycerols; Klebsiella; Klebsiella pneumoniae; Klebsiella pneumoniae - drug effects; Klebsiella pneumoniae - enzymology; Klebsiella pneumoniae - growth & development; lactates; Lactic Acid - analogs & derivatives; Lactic Acid - biosynthesis; Metabolism; Microbial Viability - drug effects; Molecular Sequence Data; Oxidoreductases - chemistry; Oxidoreductases - isolation & purification; Oxidoreductases - metabolism; Propanediol utilization protein; Propionaldehyde dehydrogenase; Recombinant; recombinant proteins; Sequence Analysis, Protein; Sirolimus - pharmacology; Strain; substrate specificity; Substrate Specificity - drug effects
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2011.09.099

► In the present work, we identified a pduP-encoded CoA-dependent propionaldehyde dehydrogenase of Klebsiella pneumoniae. ► Deletion of pduP abolished 3-HP synthesis in K. pneumoniae, especially at late stages of growth. ► Purified recombinant PduP showed broad enzymatic activity upon aliphatic aldehydes, including 3-hydroxypropionaldehyde. ► The present report is the first to confirm that the pduP gene is a member of the 3-HP biosynthesis pathway. The pduP gene encodes a propionaldehyde dehydrogenase (PduP) was investigated for the role in 3-hydroxypropionic acid (3-HP) glycerol metabolism in Klebsiella pneumoniae. The enzyme assay showed that cell extracts from a pduP mutant strain lacked measurable dehydrogenase activity. Additionally, the mutant strain accumulated the cytotoxic intermediate metabolite 3-hydroxypropionaldehyde (3-HPA), causing both cell death and a lower final 3-HP titer. Ectopic expression of pduP restored normal cell growth to mutant. The enzymatic property of recombinant protein from Escherichia coli was examined, exhibiting a broad substrate specificity, being active on 3-HPA. The present work is thus the first to demonstrate the role of PduP in glycerol metabolism and biosynthesis of 3-HP.