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Autotransporter mediated esterase display on Zymomonas mobilis and Zymobacter palmae

Paper ID Volume ID Publish Year Pages File Format Full-Text
23019 43409 2014 8 PDF Available
Title
Autotransporter mediated esterase display on Zymomonas mobilis and Zymobacter palmae
Abstract

•The E. coli autodisplay system was examined in Z. mobilis and Z. palmae.•B. gladioli EstA was displayed as an autotransporter passenger on both cell surfaces.•In both applications, EstA was proven to be enzymatically active.

Autodisplay, i.e. surface expression of recombinant proteins by virtue of the autotransporter secretion pathway, has been used predominantly with Escherichia coli as host organism, which often limits the applicability of this technique to laboratory purposes and scales. The aim of this study was to investigate if the fermentative bacteria Zymomonas mobilis and Zymobacter palmae, representing attractive candidates for industrial applications, can serve as host organisms for autodisplay. We therefore used the carboxylesterase EstA from Burkholderia gladioli as an autotransporter passenger to display it on the surfaces of Z. palmae and Z. mobilis. Expression and outer membrane localization of the EstA-autotransporter fusion protein were verified by SDS-PAGE, and surface display of the enzyme was demonstrated by ELISA and flow cytometer analysis. Whole-cell activity assays revealed that EstA retained its activity on the cell surface. Recombinant Z. palmae cells exhibited significant higher esterase activity (294 mU/mL/OD 1) in comparison to Z. mobilis (88 mU/mL/OD 1) and the control E. coli (113 mU/mL/OD 1). This appears even more noteworthy, as about 30% of EstA was released from the cell surface of Z. palmae. Nevertheless, our results indicate that both species are suitable autodisplay hosts, in particular Z. palmae for displaying esterase, opening up new horizons for biocatalytic applications.

Keywords
Zymomonas mobilis; Zymobacter palmae; Autodisplay; Esterase; Biocatalysis
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Autotransporter mediated esterase display on Zymomonas mobilis and Zymobacter palmae
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 191, 10 December 2014, Pages 228–235
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us