SCREENING STRATEGY FOR HIGH THROUGHPUT SELECTION OF NITRILASE PRODUCING MICROORGANISMS AND MUTANTS FOR THE PRODUCTION OF PHARMACEUTICALLY IMPORTANT DRUGS AND DRUG INTERME
Journal Title: International Journal of Pharmaceutical Science and Technology - Year 2011, Vol 6, Issue 1
Abstract
The prerequisite for fully exploiting the potential of a particular biocatalyst and for the laboratory evolution studies is the requirement of a reliable and convenient high throughput screening method. The major hurdle in the quest of finding novel nitrilase activities from nature as well as via directed evolution is the lack of a robust highthroughput assay for nitrilases. In the present study, we propose a ‘dual catalyst’ approach amenable to easy and rapid screening of nitrilases which is a group of enzyme having a significant role in production of various drugs/ drug intermediates like atorvastatin, ampicillin and mandelic acid. The assay is a modification to Berthelot assay. The present strategy may facilitate the screening of novel nitrilase producing microorganisms because of its ability to detect the ammonia released during the reaction at femtogram level and thus it is highly sensitive. It can even allow the selection of superior nitrilase in 96 well plate system, from a nitrilase variant library generated via directed evolution for the enhanced production of drug/ drug intermediates. Thus the present strategy promises the development of industrially viable process based on evolved nitrilases for production of enantiopure drug molecules
Authors and Affiliations
Shubhangi Kaushik , Sonali Rohamare , Utpal Mohan , U. C. Banerjee*
Keywords
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