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Improvement of Cephalosporin C Production by Recombinant DNA Integration in Acremonium chrysogenum

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Abstract

Cephalosporins are widely used as anti-infectious β-lactam antibiotics in clinic. For the purpose of increasing the yield of cephalosporin C (CPC) fermentation, especially in an industrial strain, A. chrysogenum genes cefEF and cefG, which encode the ultimate and penultimate steps in CPC biosynthesis, cefT, which encodes a CPC efflux pump, and vgb, which encodes a bacterial hemoglobin gene were transformed in various combinations into an industrial strain of A. chrysogenum. Both PCR and Southern blotting indicated that the introduced genes were integrated into the chromosome of A. chrysogenum. Carbon monoxide difference spectrum absorbance assay was performed and the result showed that Vitreoscilla hemoglobin was successfully expressed in A. chrysogenum and had biological activity. HPLC analysis of fermentation broth of recombinant A. chrysogenum showed that most transformants had a higher CPC production level than the parental strain. Multiple transformants containing an additional copy of cefG showed a significant increase in CPC production. However, cefT showed little effect on CPC production in this high producer. The highest improvement of CPC titer was observed in the mutant with an extra copy of cefG + cefEF + vgb whose CPC production was increased by 116.3%. This was the first report that three or more genes were introduced simultaneously into A. chrysogenum. Our results also demonstrated that the combination of these genes had a synergy effect in a CPC high producer.

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Acknowledgments

This study was supported by the National High-Tech Research & Development (863) Plan of China (Grant No. 2006AA020302) and by the Science and Technology Commission of Shanghai Municipality (Grant Nos. 07DZ22002, 08DZ2230600, and 08DZ220700).

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Authors and Affiliations

  1. Shanghai Institute of Pharmaceutical Industry, State Key Laboratory of New Drug and Pharmaceutical Process, 1111 Zhongshan Road (North No. 1), 200437, Shanghai, China

    Yan Liu, Guihua Gong, Liping Xie, Ning Yuan, Chunbao Zhu, Baoquan Zhu & Youjia Hu

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  1. Yan Liu
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  2. Guihua Gong
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  3. Liping Xie
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  4. Ning Yuan
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  5. Chunbao Zhu
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  6. Baoquan Zhu
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  7. Youjia Hu
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Correspondence to Youjia Hu.

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Liu, Y., Gong, G., Xie, L. et al. Improvement of Cephalosporin C Production by Recombinant DNA Integration in Acremonium chrysogenum . Mol Biotechnol 44, 101–109 (2010). https://doi.org/10.1007/s12033-009-9214-4

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