Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media
Abstract
A method is disclosed for isolating and identifying modified para-nitrobenzyl esterases. These enzymes 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. 43 figs.
- Inventors:
- Issue Date:
- Research Org.:
- California Institute of Technology (CalTech), Pasadena, CA (United States)
- Sponsoring Org.:
- USDOE, Washington, DC (United States)
- OSTI Identifier:
- 644378
- Patent Number(s):
- 5741691
- Application Number:
- PAN: 8-589,893; CNN: Grant N00014-91-J-1397
- Assignee:
- California Inst. of Tech., Pasadena, CA (United States)
- DOE Contract Number:
- FG02-93CH10578
- Resource Type:
- Patent
- Resource Relation:
- Other Information: PBD: 21 Apr 1998
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 54 ENVIRONMENTAL SCIENCES; BIODEGRADATION; NITRO COMPOUNDS; ESTERASES; ENZYMATIC HYDROLYSIS; ENZYME ACTIVITY; REMEDIAL ACTION
Citation Formats
Arnold, F H, and Moore, J C. Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media. United States: N. p., 1998.
Web.
Arnold, F H, & Moore, J C. Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media. United States.
Arnold, F H, and Moore, J C. Tue .
"Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media". United States.
@article{osti_644378,
title = {Para-nitrobenzyl esterases with enhanced activity in aqueous and nonaqueous media},
author = {Arnold, F H and Moore, J C},
abstractNote = {A method is disclosed for isolating and identifying modified para-nitrobenzyl esterases. These enzymes 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. 43 figs.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Apr 21 00:00:00 EDT 1998},
month = {Tue Apr 21 00:00:00 EDT 1998}
}