Improved stability and half-life of fluorinated phosphotriesterase using Rosetta

Ching Yao Yang, P. Douglas Renfrew, Andrew J. Olsen, Michelle Zhang, Carlo Yuvienco, Richard Bonneau, Jin Kim Montclare

Research output: Contribution to journalArticle

Abstract

Recently we demonstrated that incorporating p-fluorophenylalanine (pFF) into phosphotriesterase dramatically improved folding, thereby leading to enhanced stability and function at elevated temperatures. To further improve the stability of the fluorinated enzyme, Rosetta was used to identify multiple potential stabilizing mutations. One such variant, pFF-F104A, exhibited enhanced activity at elevated temperature and maintained activity over many days in solution at room temperature. Rock solid: To improve stability, Rosetta was used to identify the fluorinated phosphotriesterase variant, pFF-F104A. Surprisingly, pFF-F104A not only exhibited enhanced activity at elevated temperatures but also maintained activity over several days.

Original languageEnglish (US)
Pages (from-to)1761-1764
Number of pages4
JournalChemBioChem
Volume15
Issue number12
DOIs
StatePublished - Aug 18 2014

Fingerprint

Phosphoric Triester Hydrolases
Half-Life
Temperature
p-Fluorophenylalanine
Enzyme Stability
Thermodynamic properties
Rocks
Mutation
Enzymes

Keywords

  • amino acids
  • biosynthesis
  • computational design
  • half-life
  • noncanonical amino acids
  • stability

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology
  • Medicine(all)

Cite this

Improved stability and half-life of fluorinated phosphotriesterase using Rosetta. / Yang, Ching Yao; Renfrew, P. Douglas; Olsen, Andrew J.; Zhang, Michelle; Yuvienco, Carlo; Bonneau, Richard; Montclare, Jin Kim.

In: ChemBioChem, Vol. 15, No. 12, 18.08.2014, p. 1761-1764.

Research output: Contribution to journalArticle

Yang, Ching Yao ; Renfrew, P. Douglas ; Olsen, Andrew J. ; Zhang, Michelle ; Yuvienco, Carlo ; Bonneau, Richard ; Montclare, Jin Kim. / Improved stability and half-life of fluorinated phosphotriesterase using Rosetta. In: ChemBioChem. 2014 ; Vol. 15, No. 12. pp. 1761-1764.
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