Relationships between conformational stabilities and surface functional properties of mutant hen egg-white lysozymes constructed by genetic engineering

Abstract

A genetic engineering approach was introduced to elucidate the relationships between conformational stabilities and surface functional properties of hen egg-white lysozyme (HEWL). To obtain the mutants haring different stabilities, amino acid replacements were carried out by site-directed mutagenesis in the cDNA of HEWL to remove the S-S bond between Cys 76 and Cys 94 (C94A) and to delete the salt linkage between Lys13 and Leu129 (K13D). The cDNAs of mutant lysozymes C94A and K13D were inserted into a yeast expression vector (PYG-100) and expressed in Saccharomyces cerevisiae