Trehalose-6-phosphate phosphatase expression and enzymatic properties of Fusariumgraminearum

• Published in: Protein expression and purification 2025-02, Vol.226, p.106619, Article 106619
• Main Authors: Zhang, Xuebiao, Chen, Le, Ni, Zhong, Xu, Chao, Wu, Qinyan, Zhuang, Yiqing
• Format: Article
• Language: English
• Subjects: Fusarium graminearum; Heterologous expression; Trehalose-6-phosphate; Trehalose-6-phosphate phosphatase
• ISSN: 1046-5928; 1096-0279; 1096-0279
• DOI: 10.1016/j.pep.2024.106619

This study presents an exhaustive characterization of the enzymatic attributes and structural properties of trehalose-6-phosphate phosphatase (TPP) derived from Fusarium graminearum. Enzyme activity was evaluated through a meticulously designed enzymatic assay. The findings indicate that the molecular weight of the enzyme is approximately 99.8 kDa, with an optimal reaction temperature and pH of 40 °C and 6.5, respectively. Magnesium ions (Mg2+) markedly enhance the enzymatic activity, resulting in a specific activity of 1.795 U/μg. Kinetic analysis revealed a Km value of 0.96 μmol/L and a Vmax of 15.79 μmol/L/min. Subsequent computational analysis elucidated the three-dimensional architecture of the enzyme and identified the binding site for the substrate trehalose-6-phosphate (T6P). T6P was found to form hydrogen bonds with TPP at residues Lys754, Arg720, His665, Glu758, and Asn756. Additionally, hydrophobic interactions were observed between T6P and residues Phe802, Ile610, Asp801, Pro752, and Gly753. The binding energy calculated for the T6P-TPP complex stood at −5.7 kcal/mol. •The enzymatic characteristics and structure of TPP in Fusarium graminearum were first studied.•The activity of the TPP was studied by enzyme-linked reaction.•It is conducive to the subsequent development of TPP-related inhibitors.