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Molecular modification of Bacillus stearothermophilus NO2 cyclodextrin glucosyltransferase and preparation of α-cyclodextrin

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Abstract

Cyclodextrin (CD) is produced by the catalysis of starch or starch derivatives by cyclodextrin glucosyltransferase (CGTase), and its yield is mainly limited by the product and reaction specificity of CGTase. In this study, we use CGTase derived from Bacillus stearothermophilus NO2, exhibiting high expression levels and good stability for molecular modification. The N353A mutant effectively decreases the hydrolysis activity, and the ratio of the kcat values (cyclization to hydrolysis activity) is 86.46, which is threefold that of the wild type. The E142P mutant effectively enhances α-CD specificity, which increases the ratio of kcat values (α-CD to β-CD formation) from 2.18 of the wild-type to 2.42. The N353A/E142P mutant weakens the hydrolysis side reaction and enhances α-CD specificity, and the proportion of α-CD products is 53.67%, which is 15.62% higher than that of the wild-type. This research focuses on CGTase reaction and product specificities, which suggest a novel method for the industrial production of α-CD.

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Acknowledgements

We thank the National Natural Science Foundation of China (31730067 and 31972032), Agricultural Independent Innovation Fund of Jiangsu Province (CX(21)3039), and Postgraduate Research and Practice Innovation Program of Jiangsu Province (KYCX21-2025) for financial supports.

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, China

    Fangyuan Zuo, Lingqia Su, Demin Kong, Lei Wang, Jing Wu & Sheng Chen

  2. School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, Wuxi, China

    Fangyuan Zuo, Lingqia Su, Demin Kong, Lei Wang, Jing Wu & Sheng Chen

  3. International Joint Laboratory On Food Safety, Jiangnan University, Wuxi, China

    Fangyuan Zuo, Lingqia Su, Demin Kong, Lei Wang, Jing Wu & Sheng Chen

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  1. Fangyuan Zuo
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  2. Lingqia Su
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Correspondence to Sheng Chen.

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Zuo, F., Su, L., Kong, D. et al. Molecular modification of Bacillus stearothermophilus NO2 cyclodextrin glucosyltransferase and preparation of α-cyclodextrin. Syst Microbiol and Biomanuf 2, 695–704 (2022). https://doi.org/10.1007/s43393-022-00099-3

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