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Protein expression and purification. 2025 Feb 4:106687. doi: 10.1016/j.pep.2025.106687 Q41.42024

Expression and biochemical characterization of a novel NAD+-dependent xylitol dehydrogenase from the plant endophytic fungus Trichoderma gamsii

来自植物内生菌木霉菌甘姆氏变种的一个新型NAD+-依赖性的木糖醇脱氢酶的表达及生物化学性质表征 翻译改进

Shuping Fei  1, Wenxiu Hu  1, Jingwen Shu  1, Ruirui Zhao  1, Jiatong Zhao  1, Mengwei Jiang  1, Wenwen Wu  1, Chaoqun Lian  2, Wanggang Tang  3

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作者单位

  • 1 Bengbu Medical University Key Laboratory of Cancer Research and Clinical Laboratory Diagnosis, School of Laboratory Medicine, Bengbu Medical University, Anhui 233030, China.
  • 2 Bengbu Medical University Key Laboratory of Cancer Research and Clinical Laboratory Diagnosis, School of Laboratory Medicine, Bengbu Medical University, Anhui 233030, China; Department of Biochemistry and Molecular Biology, School of Laboratory Medicine, Bengbu Medical University, Bengbu 233030, Anhui, China. Electronic address: lianchaoqun@bbmu.edu.cn.
  • 3 Bengbu Medical University Key Laboratory of Cancer Research and Clinical Laboratory Diagnosis, School of Laboratory Medicine, Bengbu Medical University, Anhui 233030, China; Department of Biochemistry and Molecular Biology, School of Laboratory Medicine, Bengbu Medical University, Bengbu 233030, Anhui, China. Electronic address: tangwanggang@bbmu.edu.cn.

    • DOI: 10.1016/j.pep.2025.106687 PMID: 39914789

    摘要 Ai翻译

    Xylitol dehydrogenase (XDH; EC 1.1.1.9), encoded by the XYL2 gene, is a key enzyme in the fungal xylose metabolic pathway. In this work, a putative XDH from the plant endophytic fungus Trichoderma gamsii (TgXDH) was hetero-expressed in Escherichia coli BL21(DE3), purified to the homogeneity, and biochemically characterized. Sequence analysis revealed that TgXDH is 363 amino acids long and belongs to the zinc-containing medium-chain alcohol dehydrogenase superfamily. The size-exclusion chromatography analysis and SDS-PAGE showed that the purified recombinant TgXDH had a native molecular mass of ∼155 kDa and was composed of four identical subunits of molecular mass of ∼39 kDa. The optimum temperature and pH of this enzyme were 25 °C and pH 9.5, respectively. Kinetic analysis showed that it is an NAD+-dependent enzyme that has a polyol substrate preference (based on kcat/Km) in the order xylitol > ribitol ≈ d-sorbitol. The Km values for NAD+ with these three polyols ranged from 0.23 to 0.70 mM. Moreover, TgXDH showed high substrate affinities as compared to most of its homologs. The Km values for xylitol, ribitol, and d-sorbitol were 5.23 ± 0.68 mM, 8.01 ± 1.22 mM, and 12.34 ± 1.37 mM, respectively. Collectively, the results will contribute to understanding the biochemical properties of a novel XDH from the filamentous fungi and provide a promising XDH for industrial production of ethanol.

    Keywords: Trichoderma gamsii; Xylitol dehydrogenase; enzyme kinetics; substrate specificity.

    Keywords：NAD+-dependent; xylitol dehydrogenase; Trichoderma gamsii

    关键词：NAD+依赖型; 木糖醇脱氢酶; 曲霉甘米氏菌

    Copyright © Protein expression and purification. 中文内容为AI机器翻译，仅供参考！

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    期刊名：Protein expression and purification

    缩写：PROTEIN EXPRES PURIF

    ISSN：1046-5928

    e-ISSN：1096-0279

    IF/分区：1.4/Q4

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    Expression and biochemical characterization of a novel NAD+-dependent xylitol dehydrogenase from the plant endophytic fungus Trichoderma gamsii