Functional characterization of KanP, a methyltransferase from the kanamycin biosynthetic gene cluster of Streptomyces kanamyceticus.
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.
Nepal
Keshav Kumar
KK
Institute of Biomolecule Reconstruction (IBR), Department of Pharmaceutical Engineering, SunMoon University, #100, Kalsan-ri, Tangjeong-myeon, Asan-si, Chungnam 336-708, Republic of Korea.
Yoo
Jin Cheol
JC
Sohng
Jae Kyung
JK
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D007612:Kanamycin / Q000096:biosynthesis*; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D005810:Multigene Family; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D007612:Kanamycin; D008780:Methyltransferases