Designing the substrate specificity of d-hydantoinase using a rational approach

Sang Chul Lee, Young Jung Chang, Dong Min Shin, Jieun Han, Moon Hyeong Seo, Hossein Fazelinia, Costas D. Maranas, Hak Sung Kim

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

Enzymes that exhibit superior catalytic activity, stability and substrate specificity are highly desirable for industrial applications. These goals prompted the designed substrate specificity of Bacillus stearothermophilus d-hydantoinase toward the target substrate hydroxyphenylhydantoin (HPH). Positions crucial to substrate specificity were selected using structural and mechanistic information on the structural loops at the active site. The size and hydrophobicity of the involved amino acids were rationally changed, and the substrate specificities of the designed d-Hyd mutants were investigated. As a result, M63I/F159S exhibited about 200-fold higher specificity for HPH than the wild-type enzyme. Systematic mutational analysis and computational modeling also supported the rationale used in the design.

Original languageEnglish (US)
Pages (from-to)170-175
Number of pages6
JournalEnzyme and Microbial Technology
Volume44
Issue number3
DOIs
StatePublished - Mar 5 2009

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

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