[Construction of recombinant Escherichia coli strains producing poly (4-hydroxybutyric acid) homopolyester from glucose].
The aim of this study is to construct recombinant Escherichia coli strains capable of producing poly(4-hydroxybutyric acid) homopolyester from glucose as sole carbon source. A glutamate: succinate semialdehyde transaminase gene from Escherichia coli, a glutamate decarboxylase gene from E. coli, and a 4-hydroxybutyrate dehydrogenase gene from Ralstonia eutropha were cloned by PCR and assembled onto the plasmid pKSSE5.3 which haboured the PHA synthase gene from Ralstonia eutropha and 4-hydroxybutyrate: CoA transferase from Clostridium kluyveri. The resulting plasmids were transformed into E. coli and the pathway for biosynthesis of poly(4-hydroxybutyric acid) from glucose via alpha-ketoglutarate, an intermediate in TCA cycle was established in recombinant E. coli strains. Recombinant strains synthesized the homopolyester P(4HB), when cells were cultivated in Luria-Bertani broth with glucose as carbon source. P(4HB) accumulation was enhanced up to 30% of cell dry weight, when cells were cultivated in mineral salts M9 medium plus glucose as sole carbon source with addition of yeast extract, tryptone, casein hydrolate into medium respectively.
Biology Institute of Hebei Province, Shijiazhuang 050051, China. shuishansong@hotmail.com
Shui-shan
SS
Song
Hong
H
Ma
Zhen-xian
ZX
Gao
Zhen-hua
ZH
Jia
Xia
X
Zhang
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D010957:Plasmids; D011091:Polyesters / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic
D011091:Polyesters; C107955:poly(4-hydroxybutanoate); D005947:Glucose