Abstract
We recently proposed a metabolic engineering strategy for l-ornithine production based on the hypothesis that an increased intracellular supply of N-acetylglutamate may further enhance l-ornithine production in a well-defined recombinant strain of Corynebacterium glutamicum. In this work, an argJ-deficient arginine auxotrophic mutant of C. glutamicum is suppressed by a different locus of C. glutamicum ATCC13032. Overexpression of the NCgl1469 open reading frame (ORF), exhibiting N-acetylglutamate synthase (NAGS) activity, was able to complement the C. glutamicum arginine-auxotrophic argJ strain and showed increased NAGS activity from 0.03 to 0.17 units mg−1 protein. Additionally, overexpression of the NCgl1469 ORF resulted in a 39% increase in excreted l-ornithine. These results indicate that the intracellular supply of N-acetylglutamate is a rate-limiting step during l-ornithine production in C. glutamicum.
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Acknowledgments
This research was supported by the Advanced R&D Supporting Business between Industry and University funded by the Small and Medium Business Administration, Republic of Korea, and in part by the Sangji University Research Fund 2009. The labGENE Inc. was supported by a grant from the Gangwon Technopark.
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Hwang, GH., Cho, JY. Identification of a suppressor gene for the arginine-auxotrophic argJ mutation in Corynebacterium glutamicum . J Ind Microbiol Biotechnol 37, 1131–1136 (2010). https://doi.org/10.1007/s10295-010-0760-3
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DOI: https://doi.org/10.1007/s10295-010-0760-3