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Research article

J. Microbiol. Biotechnol. 2015; 25(11): 1835-1841

Published online November 28, 2015 https://doi.org/10.4014/jmb.1507.07030

Copyright © The Korean Society for Microbiology and Biotechnology.

Bacterial Cell Surface Display of a Multifunctional Cellulolytic Enzyme Screened from a Bovine Rumen Metagenomic Resource

Kyong-Cheol Ko 1, Binna Lee 1, Dae-Eun Cheong 1, Yunjon Han 1, Jong Hyun Choi 1* and Jae Jun Song 1

Industrial Microbiology and Bioprocess Research Center, Integrated Biorefinery Research Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 580-185, Republic of Korea

Received: July 7, 2015; Accepted: August 26, 2015

Abstract

A cell surface display system for heterologous expression of the multifunctional cellulase,
CelEx-BR12, in Escherichia coli was developed using truncated E. coli outer membrane protein
C (OmpC) as an anchor motif. Cell surface expression of CelEx-BR12 cellulase in E. coli
harboring OmpC-fused CelEx-BR12, designated MC4100 (pTOCBR12), was confirmed by
fluorescence-activated cell sorting and analysis of outer membrane fractions by western
blotting, which verified the expected molecular mass of OmpC-fused CelEx-BR12 (~72 kDa).
Functional evidence for exocellulase activity was provided by enzymatic assays of whole cells
and outer membrane protein fractions from E. coli MC4100 (pTOCBR12). The stability of E. coli
MC4100 (pTOCBR12) cellulase activity was tested by carrying out repeated reaction cycles,
which demonstrated the reusability of recombinant cells. Finally, we showed that recombinant
E. coli cells displaying the CelEx-BR12 enzyme on the cell surface were capable of growth
using carboxymethyl cellulose as the sole carbon source.

Keywords: Cell surface display, Multifunctional cellulolytic enzyme, E. coli OmpC