کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2021780 1069263 2007 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Purification and characterization of carbonic anhydrase of rice (Oryza sativa L.) expressed in Escherichia coli
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Purification and characterization of carbonic anhydrase of rice (Oryza sativa L.) expressed in Escherichia coli
چکیده انگلیسی

Rice carbonic anhydrase (CA) was successfully expressed as a glutathione-S-transferase (GST) fusion protein in an Escherichia coli expression system. The optimal induction concentration of IPTG and growth temperature was found to be 1.0 mM and 28 °C. To obtain milligram amounts of homogeneous active recombinant proteins, 150 mM NaCl and Mg-ATP solution were used during the purification procedures. After improving the conditions of expression and the purification procedures, final yield of recombinant proteins was 1.3 mg/g wet cell weight after enzymatic cleavage of the GST tag, and the molecular weight was about 29 kDa. The purified protein had CO2 hydration activity, and had no detectable esterase activity in vitro. Addition of zinc improved the CO2 hydration activity of the rice CA produced by E. coli  . The effects of acetazolamide (AZ) and the anions N3-, NO3-, I−, Br−, and Cl− on CO2 hydration activity of CA were studied. AZ and N3- were found to be strong inhibitors of rice CA. The inhibitory activity of AZ and ions was in the order AZ>N3->NO3->I->Br->Cl-I->Br->Cl-.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 52, Issue 2, April 2007, Pages 379–383
نویسندگان
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