Institute of Biomolecule Reconstruction (IBR), Department of Pharmaceutical Engineering, SunMoon University, #100, Kalsan-ri, Tangjeong-myeon, Asan-si, Chungnam 336-708, Republic of Korea.
Microbiol Res. 2010 Sep 20;165(7):557-64. doi: 10.1016/j.micres.2009.11.002. Epub 2009 Dec 16.
KanP, a putative methyltransferase, is located in the kanamycin biosynthetic gene cluster of Streptomyces kanamyceticus ATCC12853. Amino acid sequence analysis of KanP revealed the presence of S-adenosyl-L-methionine binding motifs, which are present in other O-methyltransferases. The kanP gene was expressed in Escherichia coli BL21 (DE3) to generate the E. coli KANP recombinant strain. The conversion of external quercetin to methylated quercetin in the culture extract of E. coli KANP proved the function of kanP as S-adenosyl-L-methionine-dependent methyltransferase. This is the first report concerning the identification of an O-methyltransferase gene from the kanamycin gene cluster. The resistant activity assay and RT-PCR analysis demonstrated the leeway for obtaining methylated kanamycin derivatives from the wild-type strain of kanamycin producer.
坎帕（KanP）是一种假定的甲基转移酶，位于链霉菌（Streptomyces kanamyceticus ATCC12853）的卡那霉素生物合成基因簇中。对 KanP 的氨基酸序列分析表明，它存在 S-腺苷-L-甲硫氨酸结合基序，而这种基序存在于其他 O-甲基转移酶中。将 kanP 基因在大肠杆菌 BL21（DE3）中表达，生成大肠杆菌 KANP 重组菌株。大肠杆菌 KANP 培养物提取物中外源槲皮素向甲基化槲皮素的转化证明了 kanP 作为 S-腺苷-L-甲硫氨酸依赖性甲基转移酶的功能。这是首次从卡那霉素基因簇中鉴定出 O-甲基转移酶基因的报道。抗性活性测定和 RT-PCR 分析表明，从卡那霉素产生菌的野生型菌株中获得甲基化卡那霉素衍生物具有一定的可行性。
Zotero 插件