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Enzymatic Production of Enantiopure Amino Acids from Mono-substituted Hydantoin Substrates

  • Gwynneth F. Matcher
  • Rosemary A. Dorrington
  • Stephanie G. BurtonEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 794)

Abstract

Biocatalytic conversion of 5-substituted hydantoin derivatives is an efficient method for the production of unnatural enantiomerically pure amino acids. The enzymes required to carry out this hydrolysis occur in a wide variety of eubacterial species each of which exhibit variations in substrate selectivity, enantiospecificity, and catalytic efficiency. Screening of the natural environment for bacterial strains capable of utilizing hydantoin as a nutrient source (as opposed to rational protein design of known enzymes) is a cost-effective and valuable approach for isolating microbial species with novel hydantoin-hydrolysing enzyme systems. Once candidate microbial isolates have been identified, characterization and optimization of the activity of target enzyme systems can be achieved by subjecting the hydantoin-hydrolysing system to physicochemical manipulations aimed at the enzymes activity within the natural host cells, expressed in a heterologous host, or as purified enzymes. The latter two options require knowledge of the genes encoding for the hydantoin-hydrolysing enzymes. This chapter describes the methods that can be used in conducting such development of hydantoinase-based biocatalytic routes for production of target amino acids.

Key words

Hydantoinase N-carbamoylase Optically pure amino acids Biocatalytic assay Heterologous genomic library Insertional mutant library 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Gwynneth F. Matcher
    • 1
  • Rosemary A. Dorrington
    • 1
  • Stephanie G. Burton
    • 2
    • 3
    Email author
  1. 1.Department of Biochemistry, Microbiology and BiotechnologyRhodes UniversityGrahanstownSouth Africa
  2. 2.Cape Peninsula University of TechnologyBellvilleSouth Africa
  3. 3.Research and Postgraduate StudiesUniversity of PretoriaPretoriaSouth Africa

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