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Directed evolution of stereoselective enzymes based on genetic selection as opposed to screening systems

Paper ID Volume ID Publish Year Pages File Format Full-Text
22992 43409 2014 8 PDF Available
Title
Directed evolution of stereoselective enzymes based on genetic selection as opposed to screening systems
Abstract

•Directed evolution of stereoselective enzymes offers green chemistry and biotechnology a prolific source of biocatalysts.•Stereoselective enzymes are traditionally screened using low-throughput systems based on gas and liquid chromatography.•Stereoselective selection systems are an elegant yet difficult way to solve the combinatorial issue in directed evolution.•Emergent high-throughput screening systems (e.g. FACS) are promising alternatives for engineering stereoselective enzymes.•Selection systems based on protein and metabolic engineering are yet another option for evolving stereoselective enzymes.

Directed evolution of stereoselective enzymes provides a means to generate useful biocatalysts for asymmetric transformations in organic chemistry and biotechnology. Almost all of the numerous examples reported in the literature utilize high-throughput screening systems based on suitable analytical techniques. Since the screening step is the bottleneck of the overall procedure, researchers have considered the use of genetic selection systems as an alternative to screening. In principle, selection would be the most elegant and efficient approach because it is based on growth advantage of host cells harboring stereoselective mutants, but devising such selection systems is very challenging. They must be designed so that the host organism profits from the presence of an enantioselective variant. Progress in this intriguing research area is summarized in this review, which also includes some examples of display systems designed for enantioselectivity as assayed by fluorescence-activated cell sorting (FACS). Although the combination of display systems and FACS is a powerful approach, we also envision innovative ideas combining metabolic engineering and genetic selection systems with protein directed evolution for the development of highly selective and efficient biocatalysts.

Keywords
Directed evolution; Display systems; FACS; Biocatalysis; Metabolic engineering
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Directed evolution of stereoselective enzymes based on genetic selection as opposed to screening systems
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 191, 10 December 2014, Pages 3–10
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
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Full-text PDF Download
Online Support
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