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Kharel, Madan Kumar,Lee, Hei Chan,Sohng, Jae Kyung,Liou, Kwangkyoung 한국공업화학회 2002 Journal of Industrial and Engineering Chemistry Vol.8 No.5
Different medium components were screened to improve the productivity of the novel bioactive compound, cystocin from the Streptomyces sp. GC0001. Plackett and Burman statistical design was employed to screen the effective components. Finally, response surface methodology based on three factors Box-Behnken design was applied to optimize the limiting variables such as soytone, glucose and magnesium sulfate concentration. The antibiotic yield was increased accordingly with the concentration of soytone and glucose. Magnesium sulfate has vital role in productivity besides the other carbon and nitrogen sources. Pharmamedia retained the strongest negative effect for the production of antibiotic and the effect due to sucrose and calcium carbonate was minor. The optimal concentrations of medium components for the cystocin production are determined as; soytone (50 g/L), glucose (40 g/L) and magnesium sulfate (30 g/L).
Kharel, Madan K,Basnet, Devi B,Lee, Hei Chan,Liou, Kwangkyoung,Moon, Young Ho,Kim, Jae-Jong,Woo, Jin Suk,Sohng, Jae Kyung Korean Society for Molecular Biology 2004 Molecules and cells Vol.18 No.1
<P>The organization of the 2-deoxystreptamine (DOS) biosynthetic gene cluster of Micromonospora echinospora has been determined. Sequencing of a 14.04 kb-region revealed twelve open reading frames (ORFs): four putative DOS biosynthetic genes (gtmA, B, C, and D), five amino sugars biosynthetic genes (gtmE, G, H, I, and gacB), two aminoglycoside resistance genes (gtmF and J) as well as a hypothetical ORF (gacA). One of the putative DOS biosynthetic genes, gtmA, was expressed in Escherichia coli, and the purified protein was shown to convert glucose-6-phosphate (G-6-P) to 2-deoxy-scyllo-inosose (DOI), a key step in DOS biosynthesis. In addition gtmJ was expressed in Streptomyces lividans and shown to confer gentamicin resistance. Thus gtmA and gtmJ are implicated in the biosynthesis of gentamicin and in resistance to it, respectively.</P>
Identification of 2-Deoxy-scyllo-inosose Synthase in Aminoglycoside Producer Streptomyces
KHAREL, MADAN KUMAR,SUBBA, BIMALA,LEE, HEI CHAN,LIOU, KWANGKYOUNG,WOO, JIN SUK,KIM, DONG HWAN,MOON, YOUNG-HO,SOHNG, JAE KYUNG 한국미생물 · 생명공학회 2003 Journal of microbiology and biotechnology Vol.13 No.5
Although most of the DOS containing aminoglycosides are produced by Streptomyces, very little information is available about their biosynthesis. In the present paper. we report a method to isolate DO1 synthase, a key enzyme in the biosynthesis of DOS, from aminoglycoside producer Streptomyces. PCR primers based on conserved region of DO1 synthases were specific and reliable for the isolation of the biosynthetic genes of DOS containing aminoglycosides or the screening of the aminoglycoside producers. The use of DO1 synthase as a probe could save both time and cost of cloning aminoglycoside biosynthetic genes,
Kharel, Madan Kumar,Basnet, Devi Bahadur,Lee, Hei Chan,Liou, Kwangkyoung,Woo, Jin Suk,Kim, Byung-Gee,Sohng, Jae Kyung Elsevier 2004 FEMS microbiology letters Vol.230 No.2
<P><B>Abstract</B></P><P>The biosynthetic gene cluster for tobramycin, a 2-deoxystreptamine-containing aminoglycoside antibiotic, was isolated from <I>Streptomyces tenebrarius</I> ATCC 17920. A genomic library of <I>S. tenebrarius</I> was constructed, and a cosmid, pST51, was isolated by the probes based on the core regions of 2-deoxy-<I>scyllo</I>-inosose (DOI) synthase, and <SMALL>L</SMALL>-glutamine:DOI aminotransferase and <SMALL>L</SMALL>-glutamine:<I>scyllo</I>-inosose aminotransferase. Sequencing of 33.9 kb revealed 24 open reading frames (ORFs) including putative tobramycin biosynthetic genes. We demonstrated that one of these ORFs, <I>tbmA</I>, encodes DOI synthase by in vitro enzyme assay of the purified protein. The catalytic residues of TbmA and dehydroquinate synthase were studied by homology modeling. The gene cluster found is likely to be involved in the biosynthesis of tobramycin.</P>
블록체인 애플리케이션 개발을 지원하기 위한 RESTful 패브릭 브로커
Arogya Kharel,Sumin Jeong,Joonseok Park,Keunhyuk Yeom 한국차세대컴퓨팅학회 2022 한국차세대컴퓨팅학회 학술대회 Vol.2022 No.05
Over the past few years, both public and permissioned blockchain technologies have seen a rise in adoption. Blockchainbased applications are being utilized in multiple domains and industries. Among the permissioned blockchain platforms, Hyperledger Fabric has remained popular due to its feature set and active community. However, the complexity and usability issues of Hyperledger Fabric has resulted in a cumbersome blockchain network configuration. Similarly, the amount of pre-requisite knowledge is also substantial. Therefore, we propose RESTful Fabric Broker, an intermediator that facilitates easier access to the blockchain network while also increasing its overall operability and extendibility. In addition, we suggest the conceptual architecture to represent our proposed broker’s constituent. Furthermore, we show our broker’s usability and extendibility by applying it to smart contract management and chaincode installation. We expect RESTful Fabric Broker to be used by blockchain application developers for incorporation of external blockchain-related software with improvement overall usability of the blockchain network.