Alteration and modulation of protein activity by varying post-translational modification
Embodiments of the invention include methods of altering the enzymatic activity or solubility of an extremophilic enzyme or post-translationally modifying a protein of interest via using isolated or partially purified glycosyltransferases and/or post-translational modification proteins, extracts of cells comprising glycosyltransferases and/or post-translational modification proteins, and/or in cells comprising one or more glycosyltransferases and/or post-translational modification proteins.
1. A method of altering enzymatic activity of an extremophilic enzyme, the method comprising:
introducing a nucleic acid encoding a polypeptide having at least 90% identity to SEQ ID NO: 337 into Alicyclobacillus acidocaldarius or Pichia pastoris;
allowing the Alicyclobacillus acidocaldarius or Pichia pastoris to express the polypeptide;
allowing the Alicyclobacillus acidocaldarius or Pichia pastoris to glycosylate the polypeptide;
wherein the glycosylated polypeptide has altered beta xylosidase enzymatic activity relative to an non-glycosylated form of the polypeptide.
2. The method according to claim 1 , wherein the glycosylated extremophilic enzyme has altered a beta xylosidase activity compared to a non-glycosylated form of the extremophilic enzyme at a pH selected from the group consisting of a pH of less than about 5, about 3.5, and about 2 or at a temperature selected from the group consisting of less than about 90° C., about 90° C., about 80° C., about 70° C., about 60° C., and about 50° C.