title: Heterologous production of spectinomycin in Streptomyces venezuelae by exploiting the dTDP-D-desosamine pathway.
abstract: Spectinomycin is an aminoglycoside antibiotic composed of actinamine and actinospectose, which are fused together by a putative glycosyltransferase, SpcG, during spectinomycin biosynthesis. Although previous studies have revealed the involvement of SpcA (myo-inositol monophosphatase), SpcB (dehydrogenase), SpcS2 (aminotransferase), and SpcM (methyltransferase) in the biosynthesis of actinamine, heterologous biosynthesis of spectinomycin via actinospectose has not been clearly elucidated. In this study, Streptomyces venezuelae was utilized as a source of dTDP-actinospectose from the pikromycin biosynthetic desosamine sugar pathway, and a recombinant vector, pSM5, carrying spcA, spcB, spcS2, spcM, and spcG was inserted into S. venezuelae. The formation of dTDP-spectinose was suspected through the use of dehydrogenase in the S. venezuelae chromosome. Herewith, the genetically engineered strain, S. venezuelae SM5, effectively produced up to 89.2mg/L in optimized medium. However, pSM5 in S. venezuelae YJ003, a dTDP-actinospectose-deficient strain, did not produce spectinomycin. This result demonstrates the use of a dTDP-actinospectose precursor produced in the desosamine pathway for heterologous production of spectinomycin in S. venezuelae.
authors: Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction (iBR), Sun Moon University, # 100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, Republic of Korea.
authors: Janardan
authors: J
authors: Lamichhane
authors: Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction (iBR), Sun Moon University, # 100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, Republic of Korea.
authors: Amit Kumar
authors: AK
authors: Jha
authors: Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction (iBR), Sun Moon University, # 100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, Republic of Korea.
authors: Bijay
authors: B
authors: Singh
authors: Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction (iBR), Sun Moon University, # 100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, Republic of Korea.
authors: Ramesh Prasad
authors: RP
authors: Pandey
authors: Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction (iBR), Sun Moon University, # 100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, Republic of Korea. Electronic address: sohng@sunmoon.ac.kr.
authors: Jae Kyung
authors: JK
authors: Sohng
mesh_terms: D000606:Amino Sugars / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry; D019149:Bioreactors / Q000382:microbiology*; D005798:Genes, Bacterial; D005818:Genetic Engineering; D008956:Models, Chemical; D005810:Multigene Family; D000198:Spectinomycin / Q000096:biosynthesis* / Q000737:chemistry; D013302:Streptomyces / Q000145:classification / Q000235:genetics / Q000378:metabolism*
chemical_list: D000606:Amino Sugars; D000900:Anti-Bacterial Agents; C110125:desosamine; D000198:Spectinomycin
keywords: Actinamine; Actinospectose; Desosamine pathway; Streptomyces venezuelae