TY - JOUR
T1 - Multiple display of catalytic modules on a protein scaffold
T2 - Nano-fabrication of enzyme particles
AU - Heyman, Arnon
AU - Barak, Yoav
AU - Caspi, Jonathan
AU - Wilson, David B.
AU - Altman, Arie
AU - Bayer, Edward A.
AU - Shoseyov, Oded
N1 - Funding Information:
This research was supported in part by a grant from the United States–Israel Binational Science Foundation (BSF), Jerusalem, Israel.
PY - 2007/9/30
Y1 - 2007/9/30
N2 - Self assembly is a prerequisite for fabricating nanoscale structures. Here we present a new fusion protein based on the stress-responsive homo-oligomeric protein, SP1. This ring-shaped protein is a highly stable homododecamer, which can be potentially utilized to self-assemble different modules and enzymes in a predicted and oriented manner. For that purpose, a cohesin module (a component of the bacterial cellulosome) was selected, its gene fused in-frame to SP1, and the fusion protein was expressed in Escherichia coli. The cohesin module, specialized to incorporate different enzymes through specific recognition of a dockerin modular counterpart, is used to display new moieties on the SP1 scaffold. The SP1 scaffold displayed 12 active cohesin modules and specific binding to a dockerin-fused cellulase enzyme from Thermobifida fusca. Moreover, we found a significant increase in specific activity of the scaffold-displayed enzymes.
AB - Self assembly is a prerequisite for fabricating nanoscale structures. Here we present a new fusion protein based on the stress-responsive homo-oligomeric protein, SP1. This ring-shaped protein is a highly stable homododecamer, which can be potentially utilized to self-assemble different modules and enzymes in a predicted and oriented manner. For that purpose, a cohesin module (a component of the bacterial cellulosome) was selected, its gene fused in-frame to SP1, and the fusion protein was expressed in Escherichia coli. The cohesin module, specialized to incorporate different enzymes through specific recognition of a dockerin modular counterpart, is used to display new moieties on the SP1 scaffold. The SP1 scaffold displayed 12 active cohesin modules and specific binding to a dockerin-fused cellulase enzyme from Thermobifida fusca. Moreover, we found a significant increase in specific activity of the scaffold-displayed enzymes.
KW - Cohesin-dockerin recognition
KW - Nano-bioreactor
KW - Nano-fabrication
KW - SP1
KW - Scaffold
KW - Self assembly
UR - http://www.scopus.com/inward/record.url?scp=34848919535&partnerID=8YFLogxK
U2 - 10.1016/j.jbiotec.2007.07.940
DO - 10.1016/j.jbiotec.2007.07.940
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C2 - 17826857
AN - SCOPUS:34848919535
SN - 0168-1656
VL - 131
SP - 433
EP - 439
JO - Journal of Biotechnology
JF - Journal of Biotechnology
IS - 4
ER -
