Efficient production of 2-deoxy-scyllo-inosose from d-glucose by metabolically engineered recombinant Escherichia coli.
2-Deoxy-scyllo-inosose (DOI) is a six-membered carbocycle formed from d-glucose-6-phosphate catalyzed by 2-deoxy-scyllo-inosose synthase (DOIS), a key enzyme in the biosynthesis of 2-deoxystreptamine-containing aminocyclitol antibiotics. DOI is valuable as a starting material for the benzene-free synthesis of catechol and other benzenoids. We constructed a series of metabolically engineered Escherichia coli strains by introducing a DOIS gene (btrC) from Bacillus circulans and disrupting genes for phosphoglucose isomerase, d-glucose-6-phosphate dehydrogenase, and phosphoglucomutase (pgi, zwf and pgm, respectively). It was found that deletion of the pgi gene, pgi and zwf genes, pgi and pgm genes, or all pgi, zwf and pgm genes significantly improved DOI production by recombinant E. coli in 2YTG medium (3% glucose) up to 7.4, 6.1, 11.6, and 8.4 g l(-1), respectively, compared with that achieved by wild-type recombinant E. coli (1.5 g l(-1)). Moreover, E. coli mutants with disrupted pgi, zwf and pgm genes showed strongly enhanced DOI productivity of up to 29.5 g l(-1) (99% yield) in the presence of mannitol as a supplemental carbon source. These results demonstrated that DOI production by metabolically engineered recombinant E. coli may provide a novel, efficient approach to the production of benzenoids from renewable d-glucose.
Kogure
Takahisa
T
Faculty of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Higashijima 265-1, Niitsu, Niigata 956-8603, Japan.
Wakisaka
Naoki
N
Takaku
Hiroaki
H
Takagi
Masamichi
M
D001407:Bacillus / Q000235:genetics; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D005818:Genetic Engineering / Q000379:methods; D005947:Glucose / Q000378:metabolism; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D007294:Inositol / Q000031:analogs & derivatives* / Q000096:biosynthesis; D008190:Lyases / Q000235:genetics; D010733:Phosphoglucomutase / Q000235:genetics
D007294:Inositol; C075548:2-deoxyinosose; D005954:Glucosephosphate Dehydrogenase; D008190:Lyases; C085631:2-deoxy-scyllo-inosose synthase; D005956:Glucose-6-Phosphate Isomerase; D010733:Phosphoglucomutase; D005947:Glucose