Affordable Access

deepdyve-link deepdyve-link
Publisher Website

In vivo enzyme immobilization by inclusion body display.

Authors
  • Steinmann, Björn
  • Christmann, Andreas
  • Heiseler, Tim
  • Fritz, Janine
  • Kolmar, Harald
Type
Published Article
Journal
Applied and Environmental Microbiology
Publisher
American Society for Microbiology
Publication Date
Aug 01, 2010
Volume
76
Issue
16
Pages
5563–5569
Identifiers
DOI: 10.1128/AEM.00612-10
PMID: 20581198
Source
Medline
License
Unknown

Abstract

A novel strategy for in vivo immobilization of enzymes on the surfaces of inclusion bodies has been established. It relies on expression in Escherichia coli of the polyhydroxybutyrate synthase PhaC from Cupriavidus necator, which carries at its amino terminus an engineered negatively charged alpha-helical coil (Ecoil) and forms inclusion bodies upon high-level expression. Coexpression in the same cell of galactose oxidase (GOase) from Fusarium spp. carrying a carboxy-terminal positively charged coil (lysine-rich coil [Kcoil]) sequence results in heterodimeric coiled-coil formation in vivo and in the capture of the enzyme in active form on the surface of the inclusion body particle. These round-shaped enzyme-decorated microparticles, with sizes of approximately 0.7 mum, can be isolated from lysed cells simply by centrifugation. The cost-effective one-step generation and isolation of enzymes immobilized on inclusion body particles may become useful for various applications in bioprocessing and biotransformation.

Report this publication

Statistics

Seen <100 times