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Mol. Cells 2009; 27(5): 601-608
Published online May 31, 2009
https://doi.org/10.1007/s10059-009-0080-5
© The Korean Society for Molecular and Cellular Biology
Heterologous Production of Paromamine in Streptomyces lividans TK24 Using Kanamycin Biosynthetic Genes from Streptomyces kanamyceticus ATCC12853
The 2-deoxystreptamine and paromamine are two key intermediates in kanamycin biosynthesis. In the present study, pSK-2 and pSK-7 recombinant plasmids were constructed with two combinations of genes: kanABK, and kanABKF and kacA respectively from kanamycin producer Streptomyces kanamyceticus ATCC12853. These plasmids were heterologously expressed into Streptomyces lividans TK24 independently and generated two recombinant strains named S. lividans SK-2/SL and S. lividans SK-7/SL, respectively. ESI/ MS and ESI-LC/MS analysis of the metabolite from S. lividans SK-2/SL showed that the compound had a molecular mass of 163 [M + H]+, which corresponds to that of 2-deoxystreptamine. ESI/MS and MS/MS analysis of me-tabolites from S. lividans SK-7/SL demonstrated the production of paromamine with a molecular mass of 324 [M + H]+. In this study, we report the production of paromamine in a heterologous host for the first time. This study will evoke to explore complete biosynthetic pathways of kanamycin and related aminoglycoside antibiotics.
Keywords aminoglycosides, heterologous expression, kanamycin, paromamine, Streptomyces kanamyceticus
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Research Article
Mol. Cells 2009; 27(5): 601-608
Published online May 31, 2009 https://doi.org/10.1007/s10059-009-0080-5
Copyright © The Korean Society for Molecular and Cellular Biology.
Heterologous Production of Paromamine in Streptomyces lividans TK24 Using Kanamycin Biosynthetic Genes from Streptomyces kanamyceticus ATCC12853
Keshav Kumar Nepal, Tae-Jin Oh, and Jae Kyung Sohng
Abstract
The 2-deoxystreptamine and paromamine are two key intermediates in kanamycin biosynthesis. In the present study, pSK-2 and pSK-7 recombinant plasmids were constructed with two combinations of genes: kanABK, and kanABKF and kacA respectively from kanamycin producer Streptomyces kanamyceticus ATCC12853. These plasmids were heterologously expressed into Streptomyces lividans TK24 independently and generated two recombinant strains named S. lividans SK-2/SL and S. lividans SK-7/SL, respectively. ESI/ MS and ESI-LC/MS analysis of the metabolite from S. lividans SK-2/SL showed that the compound had a molecular mass of 163 [M + H]+, which corresponds to that of 2-deoxystreptamine. ESI/MS and MS/MS analysis of me-tabolites from S. lividans SK-7/SL demonstrated the production of paromamine with a molecular mass of 324 [M + H]+. In this study, we report the production of paromamine in a heterologous host for the first time. This study will evoke to explore complete biosynthetic pathways of kanamycin and related aminoglycoside antibiotics.
Keywords: aminoglycosides, heterologous expression, kanamycin, paromamine, Streptomyces kanamyceticus
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