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Effect of Overexpression of a Soluble Pyridine Nucleotide Transhydrogenase (UdhA) on the Production of Poly(3‐hydroxybutyrate) in Escherichia coli
Author(s) -
Sánchez Ailen M.,
Andrews Jared,
Hussein Insiya,
Bennett George N.,
San KaYiu
Publication year - 2006
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp050375u
Subject(s) - escherichia coli , operon , nad+ kinase , yield (engineering) , cofactor , nucleotide , plasmid , biochemistry , metabolic engineering , strain (injury) , chemistry , polyhydroxybutyrate , biology , bacteria , enzyme , gene , materials science , genetics , anatomy , metallurgy
A soluble pyridine nucleotide transhydrogenase (UdhA) has been used to increase the productivity and yield of PHB in vivo. By inducing a high level of UdhA, which can transfer reducing equivalents between NAD and NADP, we have increased NADPH availability, resulting in high yield and productivity of PHB in Escherichia coli . Coexpression of the phb operon from Alcaligenes eutrophus H16 and the native udhA from E. coli from high copy plasmids resulted in an increase in PHB yield from 49 to 66% g of PHB per gram of total cell dry weight and an increase in final concentration from 3.52 to 6.42 g/L; the PHB concentration of the udhA carrying strain is almost twice that of the control strain expressing only the phb operon. The results of this study demonstrate the effectiveness of cofactor manipulation and its application as a tool in metabolic engineering.