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Updated: Jun 25, 2026

Functional Complementation Analysis (FCA): A Laboratory Exercise Designed and Implemented to Supplement the Teaching of Biochemical Pathways
Published on: June 24, 2016
Functional characterization of kanB by complementing in engineered Streptomyces fradiae Deltaneo6::tsr
Hum Nath Jnawali1, Bimala Subba, Kwangkyoung Liou
1Institute of Biomolecule Reconstruction, Department of Pharmaceutical Engineering, SunMoon University, 100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, Korea.
Abstract:
A putative aminotransferase gene, kanB, lies in the biosynthetic gene cluster of Streptomyces kanamyceticus ATCC 12853 and has 66% identity with neo6 in neomycin biosynthesis. Streptomyces fradiae Deltaneo6::tsr was generated by disrupting neo6 in the neomycin producer Streptomyces fradiae. Neomycin production was completely abolished in the disruptant mutant but was restored through self-complementation of neo6. S. fradiae HN4 was generated through complementation with kanB in Streptomyces fradiae neo6::tsr. Based on metabolite analysis by ESI/MS and LC/MS, neomycin production was restored in Streptomyces fradiae HN4. Thus, like neo6, kanB also functions as a 2-deoxy-scyllo-inosose aminotransferase that has dual functions in the formation of 2-deoxy-scyllo-inosose (DOS).
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