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Applied and Environmental Microbiology, June 2005, p. 3369-3372, Vol. 71, No. 6
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.6.3369-3372.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Large-Scale Engineering of the Corynebacterium glutamicum Genome

Nobuaki Suzuki,1 Satoshi Okayama,1,2 Hiroshi Nonaka,1 Yota Tsuge,1,2 Masayuki Inui,1 and Hideaki Yukawa1,2*

Microbiology Research Group, Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, Kizu-Cho, Soraku-Gun, Kyoto 619-0292, Japan,1 Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara 630-0101, Japan2

Received 28 October 2004/ Accepted 21 December 2004

The engineering of Corynebacterium glutamicum is important for enhanced production of biochemicals. To construct an improved C. glutamicum genome, we developed a precise genome excision method based on the Cre/loxP recombination system and successfully deleted 11 distinct genomic regions identified by comparative analysis of C. glutamicum genomes. Despite the loss of several predicted open reading frames, the mutant cells exhibited normal growth under standard laboratory conditions. With a total of 250 kb (7.5% of the genome), the 11 genomic regions were loaded with cryptic prophages, transposons, and genes of unknown function which were dispensable for cell growth, indicating recent horizontal acquisitions to the genome. This provides an interesting background for functional genomic studies and can be used in the improvement of cell traits.

* Corresponding author. Mailing address: Microbiology Research Group, Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, Kizu-Cho, Soraku-Gun, Kyoto 619-0292, Japan. Phone: 81-774-75-2308. Fax: 81-774-75-2321. E-mail: mmg-lab{at}rite.or.jp.

Applied and Environmental Microbiology, June 2005, p. 3369-3372, Vol. 71, No. 6
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.6.3369-3372.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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