Surface display of recombinant proteins on Escherichia coli by BclA exosporium of Bacillus anthracis.
BACKGROUND
The anchoring motif is one of the most important aspects of cell surface display as well as efficient and stable display of target proteins. Thus, there is currently a need for the identification and isolation of novel anchoring motifs.


RESULTS
A system for the display of recombinant proteins on the surface of Escherichia coli was developed using the Bacillus anthracis exosporal protein (BclA) as a new anchoring motif. For the surface display of recombinant proteins, the BAN display platform was constructed in which a target protein is linked to the C-terminus of N-terminal domain (21 amino acids) of BclA. The potential application of BAN platform for cell surface display was demonstrated with two model proteins of different size, the Bacillus sp. endoxylanase (XynA) and monooxygenase (P450 BM3m2). Through experimental analysis including outer membrane fractionation, confocal microscopy and activity assay, it was clearly confirmed that both model proteins were successfully displayed with high activities on the E. coli cell surface.


CONCLUSIONS
These results of this study suggest that the strategy employing the B. anthracis BclA as an anchoring motif is suitable for the display of heterologous proteins on the surface of E. coli and consequently for various biocatalytic applications as well as protein engineering.
BioProcess Engineering Research Center, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea. kjjeong@kaist.ac.kr.
Tae Jung
TJ
Park
Nam Su
NS
Heo
Sung Sun
SS
Yim
Jong Hyun
JH
Park
Ki Jun
KJ
Jeong
Sang Yup
SY
Lee
D001408:Bacillus anthracis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; C470593:BclA protein, Bacillus anthracis; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins