Fluorinated chloramphenicol acetyltransferase thermostability and activity profile

Improved thermostability by a single-isoleucine mutant

Natalya Voloshchuk, Man Xia Lee, Wan Wen Zhu, Ismet Caglar Tanrikulu, Jin Montclare

Research output: Contribution to journalArticle

Abstract

A lysate-based thermostability and activity profile is described for chloramphenicol acetyltransferase (CAT) expressed in trifluoroleucine, T (CAT T). CAT and 13 single-isoleucine CAT mutants were expressed in medium supplemented with T and assayed for thermostability on cell lysates. Although fluorinated mutants, L82I T and L208I T, showed losses in thermostability, the L158I T fluorinated mutant demonstrated an enhanced thermostability relative to CAT T. Further characterization of L158I T suggested that T at position 158 contributed to a portion of the observed loss in thermostability upon global fluorination.

Original languageEnglish (US)
Pages (from-to)5907-5911
Number of pages5
JournalBioorganic and Medicinal Chemistry Letters
Volume17
Issue number21
DOIs
StatePublished - Nov 1 2007

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Chloramphenicol O-Acetyltransferase
Isoleucine
Fluorination
Halogenation

Keywords

  • Mutagenesis
  • Non-natural amino acid
  • Protein engineering
  • Thermostability
  • Trifluoroleucine

ASJC Scopus subject areas

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

Cite this

Fluorinated chloramphenicol acetyltransferase thermostability and activity profile : Improved thermostability by a single-isoleucine mutant. / Voloshchuk, Natalya; Lee, Man Xia; Zhu, Wan Wen; Tanrikulu, Ismet Caglar; Montclare, Jin.

In: Bioorganic and Medicinal Chemistry Letters, Vol. 17, No. 21, 01.11.2007, p. 5907-5911.

Research output: Contribution to journalArticle

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AU - Montclare, Jin

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