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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. coli BL21 (DE3) and its mutant E. coliΔpgi successively and together so as to get the desired product. After fermentation the products were extracted and analyzed by TLC and HPLC-MS.

키워드

저자

• Nagendra Prasad Kurumbang [ Institute of Biomolecules Reconstruction, Sun Moon University ]
• Tae Jin Oh [ Institute of Biomolecules Reconstruction, Sun Moon University ]
• Kwangkyoung Liou [ Institute of Biomolecules Reconstruction, Sun Moon University ]
• Hei Chan Lee [ Institute of Biomolecules Reconstruction, Sun Moon University ]
• Jae Kyung Sohng [ Institute of Biomolecules Reconstruction, Sun Moon University ]

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간행물

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• 간행물명

  한국당과학회 학술대회

• 간기

  연간

• 수록기간

  2006~2022

• 십진분류

  KDC 517 DDC 614