Optimization of expression of human sulfite oxidase and its molybdenum domain

  • Temple C
  • Graf T
  • Rajagopalan K
  • 29

    Readers

    Mendeley users who have this article in their library.
  • 93

    Citations

    Citations of this article.

Abstract

The conditions for the heterologous expression of both untagged and His-tagged human sulfite oxidase in Escherichia coli have been optimized. Maximum production of active enzyme requires expression in a mob-cell strain at low levels of the inducer. Using these conditions, 3.9-5.6 mg of untagged and 15 mg of His-tagged sulfite oxidase were isolated per liter of cell culture. These represent significantly higher levels than previously reported for any molybdopterin-containing protein. High levels of enzyme activity and molybdenum incorporation were maintained despite the increase in yield, and no significant differences in kinetic properties were observed between the tagged and untagged sulfite oxidase. Additionally, the molybdenum domain of sulfite oxidase was expressed in a stable, active form as a His-tagged protein. The molybdenum domain was also expressed in the presence of tungstate to enable examination of the molybdopterin-tungsten form of sulfite oxidase. (C) 2000 Academic Press.

Author-supplied keywords

  • Mob
  • Molybdenum
  • Molybdenum domain
  • Molybdopterin
  • Sulfite oxidase
  • Tungsten

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Authors

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free