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A novel process using immobilized engineered strain HC01 cells with co-expressed D-hydantoinase and D-N-carbamoylase as biocatalyst for the production of D-Valine

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Abstract

The demand for D-Valine increases because of its wide range of use. A whole-cell biocatalyst for the production of D-Valine from 5′-isopropyl hydantoin by co-expression of the D-hydantoinase (hyd) gene from Pseudomonas putida YZ-26 and D-N-carbamoylase (cab) gene from Sinorhizobium sp. SS-ori in Escherichia coli BL 21 (DE3) was developed. The expression condition of the engineered strain HC01 co-expressing D-hydantoinase (HYD) and D-N-carbamoylase (CAB) was optimized. HYD and CAB reached the highest activities (4.65 and 0.75 U/ml-broth) after inducing for 8–12 h. Subsequently, the cells of HC01 were immobilized in the form of Ca2+-alginate beads, and the optimal conditions for immobilizing were obtained as 2.5% gel concentration and 0.029 g/mL cell concentration in the presence of 3% CaCl2. The thermostability of immobilized cells was 5 ℃ higher than that of free cells in the same condition. And the divalent metal ions such as Mn2+, Mg2+, Cu2+, Co2+, and Ni2+ did not significantly affect the enzymatic activity of HYD and CAB in immobilized cells. Bioconversion rate reached to 91% after a 42-h reaction when the substrate concentration was 50 mmol/L with the initial pH of 9.0 under the nitrogen protection. This method provides D-Valine with optical purity of 97% and an overall yield of 72%. Furthermore, the immobilized cells can be reused for more than 7 cycles and maintain their capacity of over 70%. Hence the immobilized cells of engineered strain HC01 could potentially be used to prepare D-Valine.

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The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

Supported by Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi (STIP) (No. 20110001) and the “1331” project in Shanxi Province.

Funding

The “1331” project in Shanxi Province, scientific and technological innovation programs of higher education institutions in Shanxi (STIP), 20110001.

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Authors and Affiliations

  1. Institute of Biotechnology, Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, 030006, Shanxi, China

    Lixi Niu, Lulu Xie, Jiao Ji & Lihua Lv

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  1. Lixi Niu
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  2. Lulu Xie
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  4. Lihua Lv
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Contributions

LN: Resources, Conceptualization, Writing – review & editing. LX: Formal analysis, Writing – original draft, Visualization. JJ: Formal analysis, Writing – original draft, Investigation. LL: Resources, Conceptualization. All authors read and approved the final manuscript.

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Correspondence to Lixi Niu.

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Niu, L., Xie, L., Ji, J. et al. A novel process using immobilized engineered strain HC01 cells with co-expressed D-hydantoinase and D-N-carbamoylase as biocatalyst for the production of D-Valine. Bioprocess Biosyst Eng 46, 227–236 (2023). https://doi.org/10.1007/s00449-022-02825-6

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