Enhanced production of GDP-l-fucose by overexpression of NADPH regenerator in recombinant Escherichia coli

Biosynthesis of guanosine 5′-diphosphate- l -fucose (GDP- l -fucose) requires NADPH as a reducing cofactor. In this study, endogenous NADPH regenerating enzymes such as glucose-6-phosphate dehydrogenase (G6PDH), isocitrate dehydrogenase (Icd), and NADP + -dependent malate dehydrogenase (MaeB) were o...

Full description

Saved in:
Bibliographic Details
Published in:Applied microbiology and biotechnology 2011-08, Vol.91 (4), p.967-976
Main Authors: Lee, Won-Heong, Chin, Young-Wook, Han, Nam Soo, Kim, Myoung-Dong, Seo, Jin-Ho
Format: Article
Language:English
Subjects:
Acids
Agricultural biotechnology
Analysis
Bacteria
Batch culture
Biological and medical sciences
Biosynthesis
Biotechnological Products and Process Engineering
Biotechnology
Dehydrogenases
Dextrose
E coli
Enzymes
Escherichia coli
Escherichia coli - enzymology
Escherichia coli - genetics
Escherichia coli - metabolism
Ethylenediaminetetraacetic acid
Fermentation
Fundamental and applied biological sciences. Psychology
Gene Expression
Genes
Genetic Engineering
Genetic recombination
Glucose
Glucose - metabolism
Glucosephosphate Dehydrogenase - genetics
Glucosephosphate Dehydrogenase - metabolism
Guanosine Diphosphate Fucose - metabolism
Isocitrate Dehydrogenase - genetics
Isocitrate Dehydrogenase - metabolism
Life Sciences
Malate Dehydrogenase (NADP+) - genetics
Malate Dehydrogenase (NADP+) - metabolism
Metabolic Networks and Pathways - genetics
Metabolism
Methods. Procedures. Technologies
Microbial engineering. Fermentation and microbial culture technology
Microbial Genetics and Genomics
Microbiology
NADP - genetics
NADP - metabolism
Phosphates
Plasmids
Studies
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Biosynthesis of guanosine 5′-diphosphate- l -fucose (GDP- l -fucose) requires NADPH as a reducing cofactor. In this study, endogenous NADPH regenerating enzymes such as glucose-6-phosphate dehydrogenase (G6PDH), isocitrate dehydrogenase (Icd), and NADP + -dependent malate dehydrogenase (MaeB) were overexpressed to increase GDP- l -fucose production in recombinant Escherichia coli . The effects of overexpression of each NADPH regenerating enzyme on GDP- l -fucose production were investigated in a series of batch and fed-batch fermentations. Batch fermentations showed that overexpression of G6PDH was the most effective for GDP- l -fucose production. However, GDP- l -fucose production was not enhanced by overexpression of G6PDH in the glucose-limited fed-batch fermentation. Hence, a glucose feeding strategy was optimized to enhance GDP- l -fucose production. Fed-batch fermentation with a pH-stat feeding mode for sufficient supply of glucose significantly enhanced GDP- l -fucose production compared with glucose-limited fed-batch fermentation. A maximum GDP- l -fucose concentration of 235.2 ± 3.3 mg l −1 , corresponding to a 21% enhancement in the GDP- l -fucose production compared with the control strain overexpressing GDP- l -fucose biosynthetic enzymes only, was achieved in the pH-stat fed-batch fermentation of the recombinant E. coli overexpressing G6PDH. It was concluded that sufficient glucose supply and efficient NADPH regeneration are crucial for NADPH-dependent GDP- l -fucose production in recombinant E. coli .
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-011-3271-x