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Fusion of Oligopeptide to the C Terminus of α-Glucuronidase from Thermotoga maritima Improves the Catalytic Efficiency for Hemicellulose Biotransformation

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Abstract

Fusion protein combined the oligopeptide (HQAFFHA) with the C terminus of α-glucuronidase from Thermotoga maritima was produced in E. coli and purified for characterization and applications of glucuronic and glucaric acid production. The fusion protein with oligopeptide exhibited a 2.97-fold higher specific activity than individual protein. Their catalytic efficiency kcat/Km and kcat increased from 469.3 ± 2.6 s−1 (g mL−1)−1 and 62.4 ± 0.9 s−1 to 2209.5 ± 26.3 s−1 (g mL−1)−1 and 293.9 ± 4.9 s−1, respectively. Fusion protein had similar temperature and pH profiles to those without oligopeptide, but the thermal stability decreases and the pH stability shifts to alkaline. Using beech xylan hydrolysate as a substrate, the glucuronic acid yield of fusion enzyme increased by 9.94% compared with its parent at 65 °C pH 8.5 for 10 h, and can hydrolyze corn cob xylan with xylanase to obtain glucuronic acid, and can be combined with uronate dehydrogenase to obtain high-added value glucaric acid. Homologous modeling analysis revealed the factors contributing to the high catalytic efficiency of fusion enzyme. These results show that the peptide fusion strategy described here may be useful for improving the catalytic efficiency and stability of other industrial enzymes, and has great potential for producing high value-added products from agricultural waste.

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Funding

This work was supported by grants from “National Key Research and Development Project” of China (Grant No. 2019YFA0706900).

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  1. Department of Food Science and Engineering, College of Food and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, 210023, People’s Republic of China

    Hongyang Zhao, Yemin Xue, Mengke Xue, Fang Xie, Yaxian Li & Zixuan Ding

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Correspondence to Yemin Xue.

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Zhao, H., Xue, Y., Xue, M. et al. Fusion of Oligopeptide to the C Terminus of α-Glucuronidase from Thermotoga maritima Improves the Catalytic Efficiency for Hemicellulose Biotransformation. Mol Biotechnol 65, 741–751 (2023). https://doi.org/10.1007/s12033-022-00569-y

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