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Characterization of a Novel Mannose Isomerase from Stenotrophomonas rhizophila and Identification of Its Possible Catalytic Residues

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Abstract

D-Mannose has great value in the treatment of chronic diseases. D-Mannose isomerase can catalyze the bioconversion of D-fructose to D-mannose. Therefore, a novel D-mannose isomerase gene (Strh-MIase) from Stenotrophomonas rhizophila strain IS26 was expressed, purified, and characterized for the industrial production of D-mannose. The specific activities of the Strh-MIase for D-mannose and D-fructose were 437.5 ± 0.8 U/mg and 267.2 ± 0.7 U/mg. Its optimal temperature and pH were 50 °C and 7.0. The enzymatic bioconversion produced 25 g/L D-mannose from concentration D-fructose (100 g/L) in 6 h by recombinant Strh-MIase, resulting in a final yield of 25%. Sodium phosphate inhibition has little influence on D-mannose production when a high concentration of D-fructose is used as substrate. We demonstrate that the metal ions improve the efficiency of D-mannose isomerase because of the enhancement of its thermostability. Moreover, the possible catalytic residues of Strh-MIase were identified by site-directed mutagenesis.

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Acknowledgements

This work was financially supported by the Natural Science Foundation of China (No. 32001636 and 31801583) and the Natural Science Foundation of Jiangsu Province (No. BK20200594 and BK20180607).

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Authors and Affiliations

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

    Yanchang Wu, Zhaolin Huang, Wenli Zhang, Cuie Guang, Qiuming Chen & Wanmeng Mu

  2. International Joint Laboratory On Food Safety, Jiangnan University, Wuxi, 214122, China

    Wanmeng Mu

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  1. Yanchang Wu
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  2. Zhaolin Huang
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  3. Wenli Zhang
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  4. Cuie Guang
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Correspondence to Qiuming Chen.

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Wu, Y., Huang, Z., Zhang, W. et al. Characterization of a Novel Mannose Isomerase from Stenotrophomonas rhizophila and Identification of Its Possible Catalytic Residues. Mol Biotechnol 64, 650–659 (2022). https://doi.org/10.1007/s12033-021-00437-1

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