کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
69611 | 48782 | 2013 | 10 صفحه PDF | دانلود رایگان |
• We report the cloning, expression and purification of the glucose isomerase gene.
• This gene belongs to a thermophilic bacterium Anoxybacillus gonensis G2T.
• It was the first study about the glucose isomerase from genus Anoxybacillus.
• This enzyme is a thermostable glucose isomerase.
• It has relatively high catalytic activity toward the substrate d-glucose.
In the continuing search for novel enzymes suitable for the production of high fructose corn syrup (HFCS), a new glucose isomerase (GI) from the thermophile Anoxybacillus gonensis G2T is described. The gene encoding this GI (AgoG2GI) was cloned and then engineered for heterologous expression in Escherichia coli. The recombinant enzyme was purified from the heat treated cell-free extract by anion exchange chromatography followed by hydrophobic interaction chromatography. The purified enzyme showed optimal activity at 85 °C and pH 6.5. The steady state parameters of Km and kcat with d-glucose were found to be 146.08 ± 9.50 mM and 36.47 ± 2.01 (1/s), respectively. l-arabinose, d-ribose and d-mannose also served as substrates for the enzyme with comparable kinetic parameters. AgoG2GI requires the divalent cations of Co2+, Mn2+ and Mg2+ for its maximal activity and thermostability. The results reported here are indicative of a new GI with desirable kinetics and stability parameters for the efficient production of HFCS at industrial scale.
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Journal: Journal of Molecular Catalysis B: Enzymatic - Volume 97, 15 December 2013, Pages 215–224