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

doi:10.1002/cbic.201402062

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 journal › Article › peer-review

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 language	English (US)
Pages (from-to)	1761-1764
Number of pages	4
Journal	ChemBioChem
Volume	15
Issue number	12
DOIs	https://doi.org/10.1002/cbic.201402062
State	Published - Aug 18 2014

Keywords

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

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Organic Chemistry

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10.1002/cbic.201402062

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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.",

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AU - Yang, Ching Yao

AU - Renfrew, P. Douglas

AU - Olsen, Andrew J.

AU - Zhang, Michelle

AU - Yuvienco, Carlo

AU - Bonneau, Richard

AU - Montclare, Jin Kim

PY - 2014/8/18

Y1 - 2014/8/18

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AB - 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.

KW - amino acids

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KW - computational design

KW - half-life

KW - noncanonical amino acids

KW - stability

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Improved stability and half-life of fluorinated phosphotriesterase using Rosetta

Abstract

Keywords

ASJC Scopus subject areas

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