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Whole-cell catalytic synthesis of trehalose by Corynebacterium glutamicum displaying trehalose synthase on its cell surface

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Abstract

Trehalose is a disaccharide with many applications in cosmetics, refrigeration, and food. Trehalose synthase is a significant enzyme in trehalose production. Escherichia coli is usually used to express this enzyme heterologously. Since E. coli is a pathogenic strain, we chose Corynebacterium glutamicum ATCC13032 as an engineering strain in this study for food safety reasons. Because of its poor permeability, we constructed two recombinant C. glutamicum strains using two anchor proteins, PorH, and short-length NCgl1337, to anchor trehalose synthase from Streptomyces coelicolor on the cell surface and synthesize trehalose directly from maltose. Studies on enzymatic properties indicated that NCgl1337S–ScTreSK246A had better enzyme activity and thermal stability than the free enzyme. After optimizing the whole-cell transformation, the optimal transformation condition was 35 °C, pH 7.0, and OD600 of 30. Under this condition, the conversion rate of 300 g/L maltose reached 69.5% in a 5 L fermentor. The relative conversion rate was still above 75% after repeated five times.

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Funding

This work was supported by the National Natural Science Foundation of China (No. 32171471), Key Research and Development Project of Shandong Province, China (2019JZZY020605), the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, Top-notch Academic Programs Project of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, Jiangsu Province, People’s Republic of China

    Ming Fang, Qiang Wang, Zhi Wang, Ruiqi Lu, Rufan **ng, **an Zhang & Zhiming Rao

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  1. Ming Fang
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  2. Qiang Wang
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  3. Zhi Wang
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  4. Ruiqi Lu
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  5. Rufan **ng
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  6. **an Zhang
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  7. Zhiming Rao
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Contributions

ZMR and XZ designed the research; MF performed the major experiments and wrote the manuscript draft; and MF, QW, ZW, RQL, RFX analyzed the data and drafted and revised the manuscript. All the authors read and approved the final manuscript.

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Correspondence to **an Zhang or Zhiming Rao.

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Fang, M., Wang, Q., Wang, Z. et al. Whole-cell catalytic synthesis of trehalose by Corynebacterium glutamicum displaying trehalose synthase on its cell surface. Syst Microbiol and Biomanuf 3, 489–497 (2023). https://doi.org/10.1007/s43393-023-00175-2

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