通过在工程改造的弗氏链霉菌Deltaneo6::tsr中进行互补来对kanB进行功能表征。

Functional characterization of kanB by complementing in engineered Streptomyces fradiae Deltaneo6::tsr.

作者信息

Jnawali Hum Nath, Subba Bimala, Liou Kwangkyoung, Sohng Jae Kyung

机构信息

Institute of Biomolecule Reconstruction, Department of Pharmaceutical Engineering, SunMoon University, 100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, Korea.

出版信息

Biotechnol Lett. 2009 Jun;31(6):869-75. doi: 10.1007/s10529-009-9937-0. Epub 2009 Feb 15.

DOI:10.1007/s10529-009-9937-0

PMID:19219581

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).

摘要

一个假定的氨基转移酶基因kanB，存在于卡那霉素链霉菌ATCC 12853的生物合成基因簇中，与新霉素生物合成中的neo6具有66%的同一性。通过破坏新霉素产生菌弗氏链霉菌中的neo6，构建了弗氏链霉菌Deltaneo6::tsr。在破坏突变体中，新霉素的产生完全被消除，但通过neo6的自我互补得以恢复。通过用弗氏链霉菌neo6::tsr中的kanB进行互补，构建了弗氏链霉菌HN4。基于电喷雾电离质谱（ESI/MS）和液相色谱/质谱（LC/MS）的代谢物分析，弗氏链霉菌HN4中恢复了新霉素的产生。因此，与neo6一样，kanB也作为2-脱氧-scyllo-肌醇氨基转移酶发挥作用，在2-脱氧-scyllo-肌醇（DOS）的形成中具有双重功能。