Evaluating the ability of polyhydroxyalkanoate synthase mutants to produce P(3HB-co-3HA) from soybean oil.
Polyhydroxyalkanoate (PHA) synthase from Pseudomonas sp 61-3 (PhaC1(Ps)) is able to synthesize P(3HB-co-3HA), consisting of a 3HB unit and medium-chain-length 3HA units of 6-12 carbon atoms. Expression vectors encoding 76 PhaC1(Ps) mutants with an amino acid replacement at position 130, 325, 477 or 481 were individually introduced into Ralstonia eutropha. The mutant enzyme genes were evaluated in terms of their abilities to synthesize P(3HB-co-3HA) using soybean oil as a carbon source. 20 mutants showed significantly high accumulation levels of PHA exceeding 30 wt.-% and as high as 57 wt.-%. It was found that hydrophobic amino acids at the positions are more likely to enhance accumulation of PHA in R. eutropha.
Tsuge
Takeharu
T
Department of Innovative and Engineered Materials, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502, Japan. tsuge.t.aa@m.titech.ac.jp
Yamamoto
Tetsuya
T
Yano
Kotaro
K
Abe
Hideki
H
Doi
Yoshiharu
Y
Taguchi
Seiichi
S
D020155:3-Hydroxybutyric Acid / Q000737:chemistry / Q000378:metabolism; D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D020118:Cupriavidus necator / Q000378:metabolism; D005632:Fructose / Q000378:metabolism; D015394:Molecular Structure; D054813:Polyhydroxyalkanoates / Q000737:chemistry / Q000378:metabolism; D011108:Polymers / Q000737:chemistry / Q000378:metabolism; D013024:Soybean Oil / Q000737:chemistry / Q000378:metabolism
D054813:Polyhydroxyalkanoates; D011108:Polymers; D005632:Fructose; D002244:Carbon; D013024:Soybean Oil; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase; D020155:3-Hydroxybutyric Acid