title: Application of acetyl-CoA acetyltransferase (AtoAD) in Escherichia coli to increase 3-hydroxyvalerate fraction in poly(3-hydroxybutyrate-co-3-hydroxyvalerate).
abstract: Polyhydroxyalkanoate (PHA) is a family of biodegradable polymers, and incorporation of different monomers can alter its physical properties. To produce the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(3HB-co-3HV)) containing a high level of 3-hydroxyvalerate (3HV) by altering acetyl-CoA pool levels, we overexpressed an acetyl-CoA acetyltransferase (atoAD) in an engineered E. coli strain, YH090, carrying PHA synthetic genes bktB, phaB, and phaC. It was found that, with introduction of atoAD and with propionate as a co-substrate, 3HV fraction in PHA was increased up to 7.3-fold higher than a strain without atoAD expressed in trans (67.9 mol%). By the analysis of CoA pool concentrations in vivo and in vitro using HPLC and LC-MS, overexpression of AtoAD was shown to decrease the amount of acetyl-CoA and increase the propionyl-CoA/acetyl-CoA ratio, ultimately resulting in an increased 3HV fraction in PHA. Finally, synthesis of P(3HB-co-3HV) containing 57.9 mol% of 3HV was achieved by fed-batch fermentation of YJ101 with propionate.
authors.affiliation: Department of Bioengineering, College of Engineering, Konkuk University, Seoul, South Korea.
authors.forename: Jong-Min
authors.initials: JM
authors.lastname: Jeon
authors.affiliation: Department of Bioengineering, College of Engineering, Konkuk University, Seoul, South Korea.
authors.forename: Hyun-Joong
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Bioengineering, College of Engineering, Konkuk University, Seoul, South Korea.
authors.forename: Shashi Kant
authors.initials: SK
authors.lastname: Bhatia
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, Kwanak-Gu, 151-742, Seoul, South Korea.
authors.forename: Changmin
authors.initials: C
authors.lastname: Sung
authors.affiliation: Department of Bioengineering, College of Engineering, Konkuk University, Seoul, South Korea.
authors.forename: Hyung-Min
authors.initials: HM
authors.lastname: Seo
authors.affiliation: Department of Bioengineering, College of Engineering, Konkuk University, Seoul, South Korea.
authors.forename: Jung-Ho
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Bioengineering, College of Engineering, Konkuk University, Seoul, South Korea.
authors.forename: Hyung-Yeon
authors.initials: HY
authors.lastname: Park
authors.affiliation: Department of Bioengineering, College of Engineering, Konkuk University, Seoul, South Korea.
authors.forename: Dahye
authors.initials: D
authors.lastname: Lee
authors.affiliation: Department of Bioengineering, University of Massachusetts Dartmouth, 285 Old Westport Road, North Dartmouth, MA, 02747-2300, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Brigham
authors.affiliation: Department of Bioengineering, College of Engineering, Konkuk University, Seoul, South Korea. seokor@konkuk.ac.kr.
authors.forename: Yung-Hun
authors.initials: YH
authors.lastname: Yang
mesh_terms: D000101:Acetyl-CoA C-Acetyltransferase / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics; D010421:Pentanoic Acids / Q000378:metabolism*; D011091:Polyesters / Q000378:metabolism*; D000094122:Polyhydroxybutyrates
chemical_list: D000101:Acetyl-CoA C-Acetyltransferase; D029968:Escherichia coli Proteins; D010421:Pentanoic Acids; D011091:Polyesters; C052620:poly(3-hydroxybutyrate)-co-(3-hydroxyvalerate); D000094122:Polyhydroxybutyrates; C013056:beta-hydroxyvaleric acid
keywords: Acetate-CoA transferase; AtoAD; Bioplastic; P(3HB-co-3HV); Polyhydroxyalkanoate