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KMID : 0545120070170091538
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Journal of Microbiology and Biotechnology
2007 Volume.17 No. 9 p.1538 ~ p.1545
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Enhancement of Clavulanic Acid by Replicative and Integrative Expression of ccaR and cas2 in Streptomyces clavuligerus NRRL3585
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Hung Trinh Viet
MALLA SAILESH
Park Byoung-Chul
Liou Kwang-Kyoung
Lee Hei-Chan
Sohng Jae-Kyung
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Abstract
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Clavulanic acid (CA) is an inhibitor of β-lactamase that is produced from Streptomyces clavuligerus NRRL3585 and is used in combination with other antibiotics in clinical treatments. In order to increase the production of CA, the replicative and integrative expressions of ccaR (encoding for a specific regulator of the CA biosynthetic operon) and cas2 (encoding for the rate-limiting enzyme in the CA biosynthetic pathway) were applied. Six recombinant plasmids were designed for this study. The pIBRHL1, pIBRHL3, and pIBRHL13 were constructed for overexpression, whereas pNQ3, pNQ2, and pNQ1 were constructed for chromosomal integration with ccaR, cas2, and ccaR-cas2, respectively. All of these plasmids were transformed into S. clavuligerus NRRL3585. CA production in transformants resulted in a significantly enhanced amount greater than that of the wild type, a 2.25-fold increase with pIBRHL1, a 9.28-fold increase with pNQ3, a 5.06-fold increase with pIBRHL3, a 2.93-fold increase with pNQ2 integration, a 5.79-fold increase with pIBRHL13, and a 23.8- fold increase with pNQ1. The integrative pNQ1 strain has been successfully applied to enhance production.
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KEYWORD
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Streptomyces clavuligerus NRRL 3585, clavulanic acid, overproduction, ccaR, cas2
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