Abstract
Identification of Stable and Enantioselective Lipases for Biotechnological Applications
Highlights
What Should be Considered During Planning Enzyme Screening Projects?S CREENING methods for identification of enzymes with desired properties are important in biotechnology
The implementation of biocatalytic reactions in biotechnological processes requires enzymes with suitable properties
Even if these methods do not provide information about a desired property, they may be very useful for pre-screening of bacterial clones producing an enzymatically active protein or carrying an expression plasmid with a correctly cloned gene of interest
Summary
S CREENING methods for identification of enzymes with desired properties are important in biotechnology. Opposite to MTP assays, throughput of chromatographic, NMR and mass spectroscopy techniques is smaller; they are considered as medium throughput methods Throughput of these methods could be increased up to several thousand samples per day through parallelization and automation.[7,8] A powerful, highly sensitive and selective ultrahigh throughput method is called fluorescence activated cell sorting (FACS) and can directly (I.e. without the need for pre-screening) be used for screening. An activityindependent screen based on processing of colonies grown on agar plates was developed allowing for the identification of protein variants with improved thermostability.[11] For FACS and MTP assays, a fluorescence or UV/VIS active functional group is attached to the surrogate substrate to allow easy handling of assays, but the results (e.g., enantioselectivity) may not necessarily be transferable to the biocatalytic conversion of “real” substrates
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