Optimization of expression of human sulfite oxidase and its molybdenum domain.

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Temple, CA; Graf, TN; Rajagopalan, KV

Published Date

  • November 15, 2000

Published In

Volume / Issue

  • 383 / 2

Start / End Page

  • 281 - 287

PubMed ID

  • 11185564

International Standard Serial Number (ISSN)

  • 0003-9861

Digital Object Identifier (DOI)

  • 10.1006/abbi.2000.2089


  • eng

Conference Location

  • United States