Expression of manganese lipoxygenase in Pichia pastoris and site-directed mutagenesis of putative metal ligands.

@article{Cristea2005ExpressionOM,
  title={Expression of manganese lipoxygenase in Pichia pastoris and site-directed mutagenesis of putative metal ligands.},
  author={Mirela Cristea and Ke Engstr{\"o}m and Chao Su and Lena H{\"o}rnsten and Ernst H. Oliw},
  journal={Archives of biochemistry and biophysics},
  year={2005},
  volume={434 1},
  pages={201-11}
}
Manganese lipoxygenase is secreted by the fungus Gaeumannomyces graminis. We expressed the enzyme in Pichia pastoris, which secreted approximately 30 mg Mn-lipoxygenase/L culture medium in fermentor. The recombinant lipoxygenase was N- and O-glycosylated (80-100 kDa), contained approximately 1 mol Mn/mol protein, and had similar kinetic properties (K(m) approximately 7.1 microM alpha-linolenic acid and V(max) 18 nmol/min/microg) as the native Mn-lipoxygenase. Mn-lipoxygenase could be… CONTINUE READING