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Targeted disruption of homoserine dehydrogenase gene and its effect on cephamycin C production in Streptomyces clavuligerus

  • Original Paper
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Journal of Industrial Microbiology & Biotechnology

Abstract

The aspartate pathway of Streptomyces clavuligerus is an important primary metabolic pathway which provides substrates for β-lactam synthesis. In this study, the hom gene which encodes homoserine dehydrogenase was cloned from the cephamycin C producer S. clavuligerus NRRL 3585 and characterized. The fully sequenced open reading frame encodes 433 amino acids with a deduced M r of 44.9 kDa. The gene was heterologously expressed in the auxotroph mutant Escherichia coli CGSC 5075 and the recombinant protein was purified. The cloned gene was used to construct a plasmid containing a hom disruption cassette which was then transformed into S. clavuligerus. A hom mutant of S. clavuligerus was obtained by insertional inactivation via double crossover, and the effect of hom gene disruption on cephamycin C yield was investigated by comparing antibiotic levels in culture broths of this mutant and in the parental strain. Disruption of hom gene resulted in up to 4.3-fold and twofold increases in intracellular free l-lysine concentration and specific cephamycin C production, respectively, during stationary phase in chemically defined medium.

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Acknowledgments

This study was supported by the Scientific and Technical Research Council of Turkey (SBAG 2753). We thank Gulveren Taskin for her technical assistance in amino acid analysis.

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Authors and Affiliations

  1. Biology Department, Middle East Technical University, Ankara, 06531, Turkey

    Ebru I. Yılmaz, Ayse K. Çaydasi & Gülay Özcengiz

  2. Biology Department, Dicle University, Diyarbakır, 21280, Turkey

    Ebru I. Yılmaz

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  1. Ebru I. Yılmaz
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  2. Ayse K. Çaydasi
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  3. Gülay Özcengiz
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Correspondence to Gülay Özcengiz.

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Yılmaz, E.I., Çaydasi, A.K. & Özcengiz, G. Targeted disruption of homoserine dehydrogenase gene and its effect on cephamycin C production in Streptomyces clavuligerus . J Ind Microbiol Biotechnol 35, 1–7 (2008). https://doi.org/10.1007/s10295-007-0259-8

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