کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8292166 1536497 2007 4 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Colorimetric endpoint assay for enzyme-catalyzed iodide ion release for high-throughput screening in microtiter plates
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Colorimetric endpoint assay for enzyme-catalyzed iodide ion release for high-throughput screening in microtiter plates
چکیده انگلیسی
Efforts are being made to engineer enzymes with enhanced activities against haloalkanes, a toxicologically important class of compounds widely used and frequently occurring in the environment. Here we describe a facile, inexpensive, and robust method for the screening of libraries of mutated enzymes with iodoalkane substrates. Iodide formed in the enzymatic reaction is oxidized to iodine, which in the presence of starch gives blue color that can be measured at 610 nm or scored with the human eye. The assay can be performed with enzymes in crude cell lysates in 96-wells microtiter plates. Expression clones of several glutathione transferases showed diverse activities with different iodoalkanes, and a mutant library of human glutathione transferase A1-1 expressed variants with enhanced substrate selectivities.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Archives of Biochemistry and Biophysics - Volume 464, Issue 2, 15 August 2007, Pages 284-287
نویسندگان
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