Abstract
We have recently developed a new l-lysine-producing mutant of Corynebacterium glutamicum by "genome breeding" consisting of characterization and reconstitution of a mutation set essential for high-level production. The strain AHP-3 was examined for l-lysine fermentation on glucose at temperatures above 35°C, at which no examples of efficient l-lysine production have been reported for this organism. We found that the strain had inherited the thermotolerance that the original coryneform bacteria was endowed with, and thereby grew and produced l-lysine efficiently up to 41°C. A final titer of 85 g/l after only 28 h was achieved at temperatures around 40°C, indicating the superior performance of the strain developed by genome breeding. When compared with the traditional 30°C fermentation, the 40°C fermentation allowed an increase in yield of about 20% with a concomitant decrease in final growth level, suggesting a significant transition of carbon flux distribution in glucose metabolism. DNA array analysis of metabolic changes between the 30°C and 40°C fermentations identified several differentially expressed genes in central carbon metabolism although we could not find stringent control-like global induction of amino-acid-biosynthetic genes in the 40°C fermentation. Among these changes, two candidates were picked out as the potential causes of the increased production at 40°C; decreased expression of the citrate synthase gene gltA and increased expression of malE, the product of which involves regeneration of pyruvate and NADPH.
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Acknowledgements
We thank Drs. S. Teshiba, A. Ozaki, and H. Anazawa for encouraging support of our work, T. Abe, S. Hashimoto, S. Koizumi, M. Yagasaki, Y. Yonetani, T. Ota, and N. Shiraishi for their useful discussions, and K. Ishimaru for his excellent technical assistance.
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Ohnishi, J., Hayashi, M., Mitsuhashi, S. et al. Efficient 40°C fermentation of l-lysine by a new Corynebacterium glutamicum mutant developed by genome breeding. Appl Microbiol Biotechnol 62, 69–75 (2003). https://doi.org/10.1007/s00253-003-1254-2
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DOI: https://doi.org/10.1007/s00253-003-1254-2