کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8331065 1540242 2015 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Study the effect of His-tag on chondroitinase ABC I based on characterization of enzyme
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Study the effect of His-tag on chondroitinase ABC I based on characterization of enzyme
چکیده انگلیسی
Chondroitinase ABC I (ChSase ABC I) which could degrade chondroitin sulfate (CS) to low molecular weight CS was expressed with His-tag in Escherichia coli (E. coli) BL21(DE3). The effect of His-tag on ChSase ABC I was investigated compared with ChSase ABC I which cut His-tag for the first time. After three steps purification, the specific activity of His-ChSase ABC I was 201.9 ± 5.4 IU/mg which was two times lower than ChSase ABC I. Results of multi angle light scattering (MALS) and analytical ultracentrifugation (AUC) showed that the polymeric state of His-ChSase ABC I was not effected by His-tag and it was monomer, and ChSase ABC I was the same. The optimal temperature and pH of His-ChSase ABC I were 37 °C and 7.5, and were almost same with ChSase ABC I. Vmax and kcat/Km of His-ChSase ABC I were 2.4 ± 0.1 μmol/L s, and 22.2 ± 0.4 L/(μmol s) and catalytic efficiency was lower than ChSase ABC I. Generally, His-tag had no effect on polymeric state, optimal temperature and pH, had little negative impact on specific activity, kcat/Km and secondary-structure of ChSase ABC I. This study might guide the application of ChSase ABC I in industrial production.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Biological Macromolecules - Volume 78, July 2015, Pages 96-101
نویسندگان
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