Open AccessInsertion of a 27 amino acid viral peptide in different zones of Escherichia coli β‐galactosidase: Effects on the enzyme activity
Author(s) -
Benito A.,
Villaverde A.
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb07208.x
Subject(s) - escherichia coli , biology , amino acid , peptide sequence , biochemistry , heterologous , hypervariable region , gene , mutant , lac operon , microbiology and biotechnology
Seven internal, putatively exposed regions of Escherichia coli β‐galactosidase have been explored regarding their tolerance to insertions of large foreign peptides. Small sequence modifications, including amino acid substitutions and small deletions, were introduced into the lacZ gene to generate unique Bam HI restriction sites. By using these mutant genes, a 27 amino acid stretch reproducing the hypervariable loop of foot‐and‐mouth disease virus VP1 protein (site A) was further inserted in predefined regions of the enzyme. Among the 13 resulting engineered proteins only three, carrying sequence modifications within a short region, are active, with only moderate reduction of their specific activities. The identified permissive region, which involves amino acids 275 to 279, seems to be a flexible area that could be appropriate incorporate and study biological properties of heterologous peptides in correctly folded β‐galactosidase chimeric proteins.