کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1985765 1540231 2016 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Epoxide hydrolase of Trichoderma reesei: Biochemical properties and conformational characterization
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Epoxide hydrolase of Trichoderma reesei: Biochemical properties and conformational characterization
چکیده انگلیسی

Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have biotechnological potential in chiral chemistry. We report the cloning, purification, enzymatic activity, and conformational analysis of the TrEH gene from Trichoderma reesei strain QM9414 using circular dichroism spectroscopy. The EH gene has an open reading frame encoding a protein of 343 amino acid residues, resulting in a molecular mass of 38.2 kDa. The enzyme presents an optimum pH of 7.2, and it is highly active at temperatures ranging from 23 to 50 °C and thermally inactivated at 70 °C (t1/2 = 7.4 min). The Michaelis constants (Km) were 4.6 mM for racemic substrate, 21.7 mM for (R)-(+)-styrene oxide and 3.0 mM for (S)-(−)-styrene oxide. The kcat/Km analysis indicated that TrEH is enantioselective and preferentially hydrolyzes (S)-(−)-styrene oxide. The conformational stability studies suggested that, despite the extreme conditions (high temperatures and extremely acid and basic pHs), TrEH is able to maintain a considerable part of its regular structures, including the preservation of the native cores in some cases. The recombinant protein showed enantioselectivity that was distinct from other fungus EHs, making this protein a potential biotechnological tool.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Biological Macromolecules - Volume 89, August 2016, Pages 569–574
نویسندگان
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