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Site-Directed Mutation of Salicylate Decarboxylase Gene and Mechanism of Ginkgo Acid Decarboxylation

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Abstract

Ginkgo seed is an important Chinese medicine and food resource in China, but the toxicity of ginkgo acid in it limits its application. Previous studies have found that salicylic acid decarboxylase (Sdc) has a decarboxylation degradation effect on ginkgo acid. In order to improve the decarboxylation ability of Sdc to Ginkgo acid, 11 residues of the Sdc around the substrate (salicylic acid) were determined as mutation targets according to the analysis of crystal structure of Sdc (PDB ID:6JQX), from Trichosporon moniliiforme WU-0401, and a total of 30 single point mutant enzymes and one compound mutant enzyme were obtained. With Ginkgo acid C15:1 as the substrate, it was found from activity assay that Sdc-Y64T and Sdc-P191A had higher decarboxylation activity, which increased by 105.18% and 116.74% compared with that of wild type Sdc, respectively. The optimal pH for Sdc Y64T and Sdc-P191A to decarboxylate Ginkgo acid C15:1 was 5.5, which is the same as the wild type Sdc. The optimal temperature of Sdc-P191A was 50 °C, which was consistent with that of the wild type Sdc, but the optimal temperature of the mutant Sdc-Y64T was 40 °C, which was 10 °C lower than that of wild type Sdc.

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Acknowledgements

We would like to appreciate Mrs. Ao He and Mr. Mian Wu, the postgraduate students of our school, for their valuable help in the experiment of this paper.

Funding

This work was supported by Natural Science Foundation of Tianjin (Grant numbers 19JCZDJC34800) and Shandong Provincial Natural Science Foundation (Project No. ZR2019MC066).

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Author notes

  1. Yuanyuan Song and Yanying Hu have contributed equally to this work.

Authors and Affiliations

  1. Tianjin Key Laboratory of Industrial Microbiology, Teda Campus, Tianjin University of Science and Technology, No. 9 of 13th Street, Tianjin Economic and Technological Development Zone TEDA, Tianjin, 300457, People’s Republic of China

    Yuanyuan Song, Jiaxin Li, Lin Wang, Wenjie Jing, Liming Zhang, Yujie Dai, Shiru Jia, Xuan Meng & Huitu Zhang

  2. Jining University, Xingtan Road, New District, Qufu City, Shandong Province, People’s Republic of China

    Yanying Hu

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  1. Yuanyuan Song
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Song, Y., Hu, Y., Li, J. et al. Site-Directed Mutation of Salicylate Decarboxylase Gene and Mechanism of Ginkgo Acid Decarboxylation. Protein J 42, 1–13 (2023). https://doi.org/10.1007/s10930-022-10086-1

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