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1.4.5.5.1 Methods for Screening and Isolation of Hydantoinases

DOI: 10.1055/sos-SD-214-00283

Slomka, C.; Engel, U.; Syldatk, C.; Rudat, J.Science of Synthesis: Biocatalysis in Organic Synthesis, (20151407.

Due to the increasing importance of hydantoinases for the industrial production of enantiomerically pure amino acids, progress has been made in finding natural sources with high hydantoinase activity. Accordingly, different screening methods have been proven to be useful. The frequently used enrichment-culture technique uses hydantoin or hydantoin derivatives as the sole nitrogen source.[‌11‌,‌157‌,‌163‌,‌164‌] In some cases, the hydantoins are utilized only during nutrient stress or catabolite repression.[‌165‌] Alternative methods to select for hydantoin-hydrolyzing strains are overlay assay on agar plates,[‌12‌] selective agar plates,[‌166‌] DNA probes,[‌167‌] as well as the utilization of polyclonal antibodies.[‌168‌] All these useful screening methods have assisted in the isolation of many organisms with hydantoinase activity. At present, high-throughput screening systems together with the directed evolution or enzyme engineering of investigated enzymes are gaining importance.

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