Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1998-04-20
1999-08-31
Prouty, Rebecca E.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
536 232, C12N 918, C12N 1555
Patent
active
059453257
ABSTRACT:
A method for isolating and identifying modified para-nitrobenzyl esterases which exhibit improved thermal stability relative to naturally occurring 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 naturally occurring 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 retain esterase activity after heat treatment at elevated temperature. Specific modified para-nitrobenzyl esterases are disclosed which have improved thermal stability and/or ester hydrolysis activity in aqueous or aqueous-organic media relative to the thermal stability and/or ester hydrolysis activity of unmodified naturally occurring para-nitrobenzyl esterase.
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Arnold Frances H.
Giver Lorraine J.
California Institute of Technology
Prouty Rebecca E.
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