Functional Analysis of Spectinomycin Biosynthetic Genes from Streptomyces spectabilis ATCC 27741

• Published in: Journal of microbiology and biotechnology Vol. 13; no. 6; pp. 906 - 911
• Main Authors: Jo, You-Young, Kim, Sun-Hee, Yang, Young-Yell, Kang, Choong-Min, Sohng, Jae-Kyung, Suh, Joo-Won
• Format: Journal Article
• Language: Korean
• Published: 한국미생물생명공학회 30.12.2003; 한국미생물·생명공학회
• Subjects: dTDP-glucose synthase; myo-inositol dehydrogenase; myo-inositol monophosphatase; Spectinomycin biosynthetic gene cluster; 생물학; Spectinomycin biosynthetic gene cluster; myo-inositol dehydrogenase; dTDP-glucose synthase; myo-inositol monophosphatase
• ISSN: 1017-7825; 1738-8872

The function of genes related to spectinomycin biosynthesis (spcD, speA, speB, spcS2) from Streptomyces spectabilis ATCC 27741, a spectinomycin producer, was analyzed. Each gene was subcloned from a spectinomycin biosynthetic gene cluster and overexpressed in E. coli BL21 (DE3) using pET vector. After incubating each purified protein with its possible substrates, the final products were analyzed using high-performance liquid chromatography (HPLC). From these results, spcD, speA, and speB have been identified to be dTDP-glucose synthase, myo-inositol monophosphatase, and myo-inositol dehydrogenase, respectively. In addition, the results suggest that the spcS2 gene product functions downstream of the speB gene product in the biosynthetic pathway of spectinomycin. Taken together, the present study elucidates the early steps of the biosynthetic pathway for 6-deoxyhexose (6-DOH) part (actinospectose) and aminocyclitol part (actinamine) of spectinomycin.