원문정보
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영어
Metabolic engineering is a powerful tool for improvement and introduction of new cellular processes in a host strain which is mostly done by genetic engineering. We constructed E. coliΔpgi mutant by disrupting the pgi and inserting neomycin/kanamycin resistant marker. The pgi encoded enzyme glucose phosphate isomerase (Pgi) is responsible for the conversion of
glucose-6-phosphate to fructose-6-phosphate and vice-versa. The mutant is a useful host for expression of a gene or gene cluster in which glucose-6-phosphate serves as a precursor. 2-deoxy-scyllo-inosose (DOI) is a first intermediate in biosynthesis of 2-deoxystreptamine
(DOS)-containing aminoglycoside such as ribostamycin, neomycin and butirosin. Subsequent amination, dehydrogenation and again amination of DOI results DOS, another stable intermediate to be isolated.
N-acetylglucosaminylation to DOS by a glycosyltransferase and deacetylation by a deacetylase gives paromamine, first and stable pseudosaccharide intermediate.
In this study five genes required for the conversion of glucose-6-phosphate to paromamine were taken from butirosin gene cluster and cloned in expression vectors. The recombinants were transformed in E. coliΔpgi successively and together so as to get the desired product. After
fermentation the products were extracted and analyzed.