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Colorimetric endpoi...
Colorimetric endpoint assay for enzyme-catalyzed iodide ion release for high-throughput screening in microtiter plates
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- Kurtovic, Sanela (författare)
- Uppsala universitet,Institutionen för biokemi och organisk kemi
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- Jansson, Ronnie (författare)
- Uppsala universitet,Institutionen för biokemi och organisk kemi
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- Mannervik, Bengt (författare)
- Uppsala universitet,Institutionen för biokemi och organisk kemi
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(creator_code:org_t)
- Elsevier BV, 2007
- 2007
- Engelska.
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Ingår i: Archives of Biochemistry and Biophysics. - : Elsevier BV. - 0003-9861 .- 1096-0384. ; 464:2, s. 284-287
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- 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. © 2007 Elsevier Inc. All rights reserved.
Ämnesord
- NATURVETENSKAP -- Kemi -- Organisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Organic Chemistry (hsv//eng)
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- Dehalogenation
- Glutathione transferase
- Iodide
- Iodoalkane
- Screening
- alkane
- glutathione transferase A1
- haloolefin
- iodine
- mutant protein
- article
- cell lysate
- colorimetry
- cost
- enzyme activity
- enzyme analysis
- enzyme engineering
- enzyme specificity
- enzyme substrate
- high throughput screening
- human
- microtiter plate assay
- nonhuman
- priority journal
- protein expression
- Enzyme Activation
- Microchemistry
- Titrimetry
- Biochemistry
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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