Incremental truncation as a strategy in the engineering of novel biocatalysts

Marc Ostermeier, Andrew E. Nixon, Stephen Benkovic

Research output: Contribution to journalArticlepeer-review

56 Scopus citations

Abstract

The application and success of combinatorial approaches to protein engineering problems have increased dramatically. However, current directed evolution strategies lack a combinatorial methodology for creating libraries of hybrid enzymes which lack high homology or for creating libraries of highly homologous genes with fusions at regions of non-identity. To create such hybrid enzyme libraries, we have developed a series of combinatorial approaches that utilize the incremental truncation of genes, gene fragments or gene libraries. For incremental truncation, Exonuclease III is used to create a library of all possible single base-pair deletions of a given piece of DNA. Incremental truncation libraries (ITLs) have applications in protein engineering as well as protein folding, enzyme evolution, and the chemical synthesis of proteins. In addition, we are developing a methodology of DNA shuffling which is independent of DNA sequence homology.

Original languageEnglish (US)
Pages (from-to)2139-2144
Number of pages6
JournalBioorganic and Medicinal Chemistry
Volume7
Issue number10
DOIs
StatePublished - Jan 1 1999

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

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