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
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موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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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
Journal: Protein Expression and Purification - Volume 77, Issue 2, June 2011, Pages 153-158
نویسندگان
Benjamin E. Lindenmuth, Karen A. McDonald,