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Combinatorial Libra...
Combinatorial Library Based Engineering of Candida antarctica Lipase A for Enantioselective Transacylation of sec-Alcohols in Organic Solvent
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- Wikmark, Ylva (författare)
- Stockholms universitet,Institutionen för organisk kemi
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- Humble, Maria Svedendahl (författare)
- KTH,Stockholms universitet,Institutionen för organisk kemi,Industriell bioteknologi,Stockholm University, Sweden,Albanova University Center
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- Bäckvall, Jan-E. (författare)
- Stockholms universitet,Institutionen för organisk kemi
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(creator_code:org_t)
- 2015-02-09
- 2015
- Engelska.
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Ingår i: Angewandte Chemie International Edition. - : Wiley. - 1433-7851 .- 1521-3773. ; 54:14, s. 4284-4288
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Abstract
Ämnesord
Stäng
- A method for determining lipase enantioselectivity in the transacylation of sec-alcohols in organic solvent was developed. The method was applied to a model library of Candida antarctica lipase A (CalA) variants for improved enantioselectivity (E values) in the kinetic resolution of 1-phenylethanol in isooctane. A focused combinatorial gene library simultaneously targeting seven positions in the enzyme active site was designed. Enzyme variants were immobilized on nickel-coated 96-well microtiter plates through a histidine tag (His6 -tag), screened for transacylation of 1-phenylethanol in isooctane, and analyzed by GC. The highest enantioselectivity was shown by the double mutant Y93L/L367I. This enzyme variant gave an E value of 100 (R), which is a dramatic improvement on the wild-type CalA (E=3). This variant also showed high to excellent enantioselectivity for other secondary alcohols tested.
Ämnesord
- NATURVETENSKAP -- Kemi -- Organisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Organic Chemistry (hsv//eng)
- NATURVETENSKAP -- Kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences (hsv//eng)
Nyckelord
- biocatalysis
- kinetic resolution
- lipase A
- protein engineering
- secondary alcohols
- organisk kemi
- Organic Chemistry
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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