title: Production of 3-hydroxypropionic acid from glycerol by recombinant Pseudomonas denitrificans.
abstract: 3-Hydroxypropionic acid (3-HP) can be produced from glycerol through two sequential enzymatic reactions that are catalyzed by a coenzyme B12 -dependent glycerol dehydratase and an NAD(P)(+) -dependent aldehyde dehydrogenase (ALDH), respectively. Pseudomonas denitrificans synthesizes coenzyme B12 under aerobic conditions, where NAD(P)(+) is regenerated efficiently. Hence, it is considered an ideal host for the production of 3-HP from glycerol under aerobic conditions. In this study, recombinant strains of P. denitrificans were developed and their potential for the production of 3-HP from glycerol was evaluated. When the enzymes, glycerol dehydratase (DhaB) and glycerol dehydratase reactivase (GdrAB), of Klebsiella pneumoniae were expressed heterologously, P. denitrificans could produce 3-HP at 37.7 mmol/L with 62% (mol/mol) yield on glycerol. Glucose was required as the carbon and energy sources for cell growth. The overexpression of heterologous ALDH was not essential; however, the titer and yield of 3-HP were improved to 54.7 mmol/L and 67% (mol/mol), respectively, when an ALDH gene (puuC) from K. pneumoniae was overexpressed. One serious drawback hindering the use of P. denitrificans as a recombinant host for 3-HP production is that it oxidizes 3-HP to malonate and utilizes 3-HP as a carbon source for growth. This is the first report on the development and use of recombinant P. denitrificans for 3-HP production from glycerol.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Pusan National University, San 30, Jangeon-dong, Geumjeong-gu, Busan, 609-735, Republic of Korea.
authors.forename: Shengfang
authors.initials: S
authors.lastname: Zhou
authors.forename: Christy
authors.initials: C
authors.lastname: Catherine
authors.forename: Chelladurai
authors.initials: C
authors.lastname: Rathnasingh
authors.forename: Ashok
authors.initials: A
authors.lastname: Somasundar
authors.forename: Sunghoon
authors.initials: S
authors.lastname: Park
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D004734:Energy Metabolism; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism*; D006836:Hydro-Lyases / Q000235:genetics / Q000378:metabolism; D019344:Lactic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D008314:Malonates / Q000378:metabolism; D060847:Metabolic Engineering; D011549:Pseudomonas / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008314:Malonates; D011994:Recombinant Proteins; D019344:Lactic Acid; D002244:Carbon; C030290:malonic acid; C031601:hydracrylic acid; D000444:Aldehyde Dehydrogenase; D006836:Hydro-Lyases; C120061:glycerol dehydratase reactivating factor, Klebsiella pneumoniae; C020671:glycerol dehydratase; D005947:Glucose; D005990:Glycerol
keywords: 3-hydroxypropionic acid; Pseudomonas denitrificans; coenzyme B12; glycerol