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Simultaneous utilization of d-cellobiose, d-glucose, and d-xylose by recombinant Corynebacterium glutamicum under oxygen-deprived conditions

  • Applied Genetics and Molecular Biotechnology
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Abstract

Corynebacterium glutamicum R was metabolically engineered to broaden its sugar utilization range to d-xylose and d-cellobiose contained in lignocellulose hydrolysates. The resultant recombinants expressed Escherichia coli xylA and xylB genes, encoding d-xylose isomerase and xylulokinase, respectively, for d-xylose utilization and expressed C. glutamicum R bglF 317A and bglA genes, encoding phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) β-glucoside-specific enzyme IIBCA component and phospho-β-glucosidase, respectively, for d-cellobiose utilization. The genes were fused to the non-essential genomic regions distributed around the C. glutamicum R chromosome and were under the control of their respective constitutive promoter trc and tac that permitted their expression even in the presence of d-glucose. The enzyme activities of resulting recombinants increased with the increase in the number of respective integrated genes. Maximal sugar utilization was realized with strain X5C1 harboring five xylA–xylB clusters and one bglF 317A–bglA cluster. In both d-cellobiose and d-xylose utilization, the sugar consumption rates by genomic DNA-integrated strain were faster than those by plasmid-bearing strain, respectively. In mineral medium containing 40 g l−1d-glucose, 20 g l−1d-xylose, and 10 g l−1d-cellobiose, strain X5C1 simultaneously and completely consumed these sugars within 12 h and produced predominantly lactic and succinic acids under growth-arrested conditions.

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Acknowledgments

We thank Crispinus A. Omumasaba (RITE) for helpful comments on the manuscript. This work was supported by a grant from the New Energy and Industrial Technology Development Organization (NEDO), Japan.

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Authors and Affiliations

  1. Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, Kizugawa-shi, Kyoto, 619-0292, Japan

    Miho Sasaki, Toru Jojima, Masayuki Inui & Hideaki Yukawa

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  1. Miho Sasaki
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  2. Toru Jojima
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  3. Masayuki Inui
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  4. Hideaki Yukawa
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Correspondence to Hideaki Yukawa.

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Sasaki, M., Jojima, T., Inui, M. et al. Simultaneous utilization of d-cellobiose, d-glucose, and d-xylose by recombinant Corynebacterium glutamicum under oxygen-deprived conditions. Appl Microbiol Biotechnol 81, 691–699 (2008). https://doi.org/10.1007/s00253-008-1703-z

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  • DOI: https://doi.org/10.1007/s00253-008-1703-z

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