Activity control of autodisplayed proteins on the same outer membrane layer of E. coli by using Z-domain/streptavidin/and lipase/foldase systems

Abstract

The autodisplay technology has been applied for expression of a desired protein on the outer membrane (OM) of Escherichia coli. In this work, the OM fractions of E. coli with two autodisplayed proteins were separately prepared and mixed to demonstrate the feasibility of control over the ratio of two autodisplayed proteins. As the first model, Z-domain and streptavidin were autodisplayed, and their activities were tested by means of the combined OM layer in a 96-well microplate and a surface plasmon resonance (SPR) biosensor. As the second model, lipase and foldase were autodisplayed which required an interaction between two proteins to obtain the activity of lipase. The OM fractions of E. coli with an autodisplayed lipase and foldase were separately prepared and mixed to demonstrate the feasibility of control over the ratio of two autodisplayed proteins when the interaction of two proteins is required within the same OM layer for the activity of the lipase. (C) 2016 Elsevier Inc. All rights reserved.