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Title: Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media

Abstract

A method for isolating and identifying modified para-nitrobenzyl esterases which exhibit improved stability and/or esterase hydrolysis activity toward selected substrates and under selected reaction conditions relative to the unmodified para-nitrobenzyl esterase. The method involves preparing a library of modified para-nitrobenzyl esterase nucleic acid segments (genes) which have nucleotide sequences that differ from the nucleic acid segment which encodes for unmodified para-nitrobenzyl esterase. The library of modified para-nitrobenzyl nucleic acid segments is expressed to provide a plurality of modified enzymes. The clones expressing modified enzymes are then screened to identify which enzymes have improved esterase activity by measuring the ability of the enzymes to hydrolyze the selected substrate under the selected reaction conditions. Specific modified para-nitrobenzyl esterases are disclosed which have improved stability and/or ester hydrolysis activity in aqueous or aqueous-organic media relative to the stability and/or ester hydrolysis activity of unmodified naturally occurring para-nitrobenzyl esterase.

Inventors:
 [1];  [1]
  1. (Pasadena, CA)
Issue Date:
Research Org.:
California Institute of Technology (Pasadena, CA)
OSTI Identifier:
871483
Patent Number(s):
5741691
Assignee:
California Institute of Technology (Pasadena, CA) CHO
DOE Contract Number:  
FG02-93CH10578
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
para-nitrobenzyl; esterases; enhanced; activity; aqueous; nonaqueous; media; method; isolating; identifying; modified; exhibit; improved; stability; esterase; hydrolysis; selected; substrates; reaction; conditions; relative; unmodified; involves; preparing; library; nucleic; acid; segments; genes; nucleotide; sequences; segment; encodes; expressed; provide; plurality; enzymes; clones; expressing; screened; identify; measuring; ability; hydrolyze; substrate; specific; disclosed; ester; aqueous-organic; naturally; occurring; selected reaction; modified enzymes; para-nitrobenzyl esterases; acid segments; hydrolysis activity; involves preparing; exhibit improved; reaction conditions; naturally occurring; nucleotide sequences; nucleotide sequence; method involves; nucleic acid; aqueous media; para-nitrobenzyl esterase; modified para-nitrobenzyl; improved stability; selected substrate; organic media; selected substrates; nonaqueous media; method involve; identifying modified; esterase hydrolysis; enhanced activity; modified enzyme; acid segment; /435/999/

Citation Formats

Arnold, Frances H., and Moore, Jeffrey C. Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media. United States: N. p., 1998. Web.
Arnold, Frances H., & Moore, Jeffrey C. Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media. United States.
Arnold, Frances H., and Moore, Jeffrey C. Thu . "Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media". United States. https://www.osti.gov/servlets/purl/871483.
@article{osti_871483,
title = {Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media},
author = {Arnold, Frances H. and Moore, Jeffrey C.},
abstractNote = {A method for isolating and identifying modified para-nitrobenzyl esterases which exhibit improved stability and/or esterase hydrolysis activity toward selected substrates and under selected reaction conditions relative to the unmodified para-nitrobenzyl esterase. The method involves preparing a library of modified para-nitrobenzyl esterase nucleic acid segments (genes) which have nucleotide sequences that differ from the nucleic acid segment which encodes for unmodified para-nitrobenzyl esterase. The library of modified para-nitrobenzyl nucleic acid segments is expressed to provide a plurality of modified enzymes. The clones expressing modified enzymes are then screened to identify which enzymes have improved esterase activity by measuring the ability of the enzymes to hydrolyze the selected substrate under the selected reaction conditions. Specific modified para-nitrobenzyl esterases are disclosed which have improved stability and/or ester hydrolysis activity in aqueous or aqueous-organic media relative to the stability and/or ester hydrolysis activity of unmodified naturally occurring para-nitrobenzyl esterase.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1998},
month = {1}
}

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