Engineering of human lysozyme as a polyelectrolyte by the alteration of molecular surface charge.
The surface positive charges of human lysozyme were either increased or decreased to alter the electrostatic interaction between enzyme and substrate in the lytic action of human lysozyme using site-directed mutagenesis. The amino acid substitutions accompanying either the addition or the removal of two units of positive charge have shifted the optimal ionic strength (NaCl concentration in 10 mM Mes buffer, pH 6.2) for the lysis of Micrococcus lysodeikticus cell from 0.04 M to 0.1 M and from 0.04 M to 0.02 M respectively. In addition to the change in ionic strength-activity profile, the pH-activity profile and the effect of a polycationic electrolyte, poly-L-Lys-HCl, on the lytic activity were significantly changed. Owing to the shifts in both ionic strength profiles and pH profiles the Arg74/Arg126 mutant has become a better catalyst than wild-type enzyme under the conditions of high ionic strength and high pH, and the Gln41/Ser101 mutant has become a better catalyst under the conditions of low ionic strength and low pH.
Muraki
M
M
National Chemical Laboratory for Industry, Ibaraki, Japan.
Morikawa
M
M
Jigami
Y
Y
Tanaka
H
H
D000595:Amino Acid Sequence; D006801:Humans; D008969:Molecular Sequence Data; D009113:Muramidase / Q000235:genetics*; D009154:Mutation; D011487:Protein Conformation; D015202:Protein Engineering; D012441:Saccharomyces cerevisiae / Q000378:metabolism
D009113:Muramidase