Applications for Bacillus megaterium in synthetic biology and modular protein and peptide secretion

Authors
Marchand, Nicholas
ORCID
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Other Contributors
Collins, Cynthia H.
Wang, Chunyu
Belfort, Georges
Karande, Pankaj
Issue Date
2015-05
Keywords
Chemical engineering
Degree
PhD
Terms of Use
This electronic version is a licensed copy owned by Rensselaer Polytechnic Institute, Troy, NY. Copyright of original work retained by author.
Full Citation
Abstract
The second half of our work describes the development and characterization of a novel system for modular protein/peptide secretion from B. megaterium. Secretion can often lead to increased stability and solubility of recombinant proteins/peptides, as well as a simplified purification scheme. As B. megaterium couples efficient protein secretion with a low-protease extracellular environment it may be an ideal host for the production of peptides, which have been difficult to produce using traditional methods because of their susceptibility to protease degradation. Recombinant proteins/peptides which are not naturally secreted can be secreted from bacterial hosts by fusing a short signal peptide (SP) taken from a naturally secreted protein to their N-terminus. However, recombinant secretion has been an underutilized tool largely due to the development time required to find a match between SP and protein that will provide for efficient secretion. The new system described here consists of a SP, Linker, and Intein (SPLInt) fused to a variable target protein or peptide. The SPLInt system provides efficient passage of a target protein/peptide outside of the cell, followed by intein-mediated cleavage to release the target for extracellular accumulation. We anticipate that these new tools will expand the applications for B. megaterium, and increase its use in both academia and industry.
Description
May 2015
School of Engineering
Department
Dept. of Chemical and Biological Engineering
Publisher
Rensselaer Polytechnic Institute, Troy, NY
Relationships
Rensselaer Theses and Dissertations Online Collection
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