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Breeding anl-isoleucine producer by protoplast fusion ofCorynebacterium glutamicum
Authors:Satoru Furukawa  Tomoki Azuma  Toshihide Nakanishi  Masahiro Sugimoto
Institution:(1) Technical Research Laboratories, Hofu Plant, Kyowa Hakko Kogyo Co., Ltd., Kyowa-machi, 747 Hofu-shi, Yamaguchi, Japan;(2) Present address: Kyowa Hakko Kogyo Co., Ltd., Ote-machi, Chiyoda-ku, 1-6-1, 100 Tokyo, Japan
Abstract:Summary Corynebacterium glutamicum R-18 is a strain forl-isoleucine production. Polyethylene glycol (PEG)-induced protoplast fusion was applied to improve the strain of thisl-isoleucine producer. Strain R-18 was fused with anl-lysine producerC. glutamicum S-37, becausel-isoleucine andl-lysine are synthesized from a common intermediate, aspartate-beta-semialdehyde. Two thousand fusants were checked for their phenotypes. Most of the fusants accumulatedl-lysine, and only 0.9% of the fusants accumulatedl-isoleucine. Two strains, F-28 and F-91, were selected and cultivated in production medium. Fusant F-28 accumulated 12.1 g/l ofl-isoleucine and 4.8 g/l ofl-lysine, and fusant F-91 accumulated 4.8 g/l ofl-isoleucine and 13.0 g/l ofl-lysine, while the parental strains R-18 and S-37 accumulated 9.5 g/l ofl-isoleucine and 26.8 g/l ofl-lysine, respectively. Sugar consumption activity was improved by protoplast fusion, and thel-isoleucine production rate of F-28 was 2.4 times higher than that of R-18.
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