Effect of Overexpression of a Soluble Pyridine Nucleotide Transhydrogenase (UdhA) on the Production of Poly(3-hydroxybutyrate) in Escherichia coli

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...

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Bibliographic Details
Published in:Biotechnology progress 2006-03, Vol.22 (2), p.420-425
Main Authors: Sánchez, Ailen M., Andrews, Jared, Hussein, Insiya, Bennett, George N., San, Ka-Yiu
Format: Article
Language:English
Subjects:
Alcaligenes eutrophus
Biological and medical sciences
Biotechnology
Escherichia coli
Escherichia coli - enzymology
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Enzymologic
Glucose - metabolism
Hydroxybutyrates - metabolism
NAD - metabolism
NADP Transhydrogenases - genetics
NADP Transhydrogenases - metabolism
Polyesters - metabolism
Solubility
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Summary: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 phboperon from Alcaligenes eutrophusH16 and the native udhAfrom 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.
ISSN:8756-7938
1520-6033
DOI:10.1021/bp050375u