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Enantioselective resolution of (R,S)-DMPM to prepare (R)-DMPM by an adsorbed-covalent crosslinked esterase PAE07 from Pseudochrobactrum asaccharolyticum WZZ003

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Abstract

(R)-N-(2,6-dimethylphenyl) aminopropionic acid methyl ester ((R)-DMPM) is an important chiral intermediate of the fungicide N-(2,6-Dimethylphenyl)-N-(methoxyacetyl)-alanine methyl ester ((R)-Metalaxyl). In this study, (1) D3520 (macroporous acrylic anion resin), selected from the ten resins, was used to immobilize the esterase from Pseudochrobactrum asaccharolyticum WZZ003 (PAE07) for resoluting the (R,S)-DMPM to obtain (R)-DMPM. (2) Up to 20 g/L PAE07 could be immobilized onto D3520 with a high enzymatic activity of 32.4 U/g. Moreover, the Km and Vmax values of 19.1 mM and 2.8 mM/min for D3520-immobilized PAE07 indicated its high activity and stereoselectivity. (3) The optimal temperature and pH for the immobilized PAE07 were 40 ℃ and 8.0, and substrate concentration was up to 0.35 M. After 15 h reaction, the conversion rate from (R,S)-DMPM to (R)-DMPM was 48.0% and the e.e.p and E values were 99.5% and 1393.0, respectively. In scale-up resolution, 200 g/L substrate and 12.5 g immobilized esterase PAE07 condition, a conversion rate from substrate to product of 48.1% and a product e.e.p of 98% were obtained within 12 h, with the activity of immobilized PAE07 retained 80.2% after 5 cycles of reactions. These results indicated that the D3520-immobilized esterase PAE07 had great potential for enzymatic resolution of (R,S)-DMPM to prepare (R)-Metalaxyl.

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The raw data used to support the findings of this study are available from the corresponding author upon request.

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Acknowledgements

The authors are thankful for Zhejiang Provincial Natural Science Foundation (LY18B020021) for financial support.

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  1. Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, No.1 Gongda Road, Deqing, Zhejiang, 313299, China

    Yin-Jun Zhang, Li-Tian Wei, Ming-Peng Zhou, Chun Wei & Xin-Jun Yu

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Contributions

Y-JZ: conceptualization, methodology, formal analysis, funding acquisition, investigation, writing-review and editing. L-TW: methodology, validation, formal analysis. M-PZ: methodology, validation, formal analysis, writing-original draft, visualization. CW: methodology, writing-review and editing, project administration. X-JY: conceptualization, writing-review and editing, project administration, supervision.

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Correspondence to Chun Wei or Xin-Jun Yu.

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Zhang, YJ., Wei, LT., Zhou, MP. et al. Enantioselective resolution of (R,S)-DMPM to prepare (R)-DMPM by an adsorbed-covalent crosslinked esterase PAE07 from Pseudochrobactrum asaccharolyticum WZZ003. Bioprocess Biosyst Eng 46, 171–181 (2023). https://doi.org/10.1007/s00449-022-02821-w

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