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The expression in E. coli of a polymeric gene coding for an esterase mimic catalyzing the hydrolysis of p ‐nitrophenyl esters
Author(s) -
Bülow Leif,
Mosbach Klaus
Publication year - 1987
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(87)81325-x
Subject(s) - hydrolysis , chemistry , esterase , moiety , bifunctional , biochemistry , galactokinase , oligonucleotide , stereochemistry , catalysis , gene , enzyme , escherichia coli
We have prepared a hybrid protein consisting of seven esterase units, Glu‐Ala‐His‐Ala‐Ser‐Phe‐Phe‐Phe, fused to the N‐terminal of galactokinase ( E. coli ). The structural gene for this bifunctional protein was obtained by cloning a polymer made up of three chemically synthesized oligonucleotides to which the galactokinase gene was fused in frame. The hybrid protein was purified to homogeneity with the aid of the galactokinase moiety and showed an M r of 51 000‐53 000. The preparation could catalyze the hydrolysis of p ‐nitrophenyl esters and, due to the inbuilt hydrophobic spacers, Phe‐Phe‐Phe, improved catalysis of more hydrophobic substrates was obtained.