کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2020776 1069209 2011 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Production and characterization of Acidothermus cellulolyticus endoglucanase in Pichia pastoris
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Production and characterization of Acidothermus cellulolyticus endoglucanase in Pichia pastoris
چکیده انگلیسی
The endoglucanase (E1) from Acidothermus cellulolyticus has been used extensively in cellulase research. The goal of this work was to produce high levels of this enzyme in a system that facilitates purification. A codon-optimized synthetic gene for A. cellulolyticus E1 with a C-terminal histidine tag was cloned into the genome of Pichia pastoris. Strain KM71H expressed the most enzyme, with a yield of 550 mg/L culture supernatant. The temperature optimum (80 °C) and pH optimum (5.1) of the purified enzyme agree with previously determined values for the enzyme produced in other systems. Michaelis-Menten kinetic parameters were determined, using a fluorescent substrate (methylumbelliferyl-β-d-cellobioside) at various temperatures. This thermostable enzyme can be used in future cellulosic biofuels-related research.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 77, Issue 2, June 2011, Pages 153-158
نویسندگان
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