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Selection and screening for enzymes of nitrile metabolism

Paper ID Volume ID Publish Year Pages File Format Full-Text
24807 43538 2008 9 PDF Available
Title
Selection and screening for enzymes of nitrile metabolism
Abstract

This work critically reviews the assays of nitrile-converting and nitrile-forming enzymes (nitrilases, nitrile hydratases, amidases, aldoxime dehydratases). Most of the strains producing such enzymes were obtained by selection on media with nitriles, amides or aldoximes as nitrogen sources. Activity and enantioselectivity of the enzymes was usually assayed by time-consuming chromatographic analysis of substrates and the corresponding reaction products. Attempts at introducing faster assays resulted in several spectrophotometric methods for reaction product (ammonia, hydroxamate, methacrylamide, benzamide, etc.) determination. Recently, new methods for colorimetric and fluorimetric determination of ammonia have been developed, which appear promising for high-throughput assays. Alternatively, methods consisting in determination of NADH consumed in a coupled amination reaction or pH-responsive methods are promising for this purpose. All the above selection and screening methods establish fundamental conditions for the design of hierarchical screening projects. However, the potential of these principles, in particular spectrophotometric and fluorimetric methods, will be probably further exploited and adapted to multiwell plate and robotic systems.

Keywords
Nitrilase; Nitrile hydratase; Amidase; Aldoxime dehydratase; Selection; Screening
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Selection and screening for enzymes of nitrile metabolism
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 133, Issue 3, 1 February 2008, Pages 318–326
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
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us