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Efficient production of extracellular proteins with Escherichia coli by means of optimized coexpression of bacteriocin release proteins

Paper ID Volume ID Publish Year Pages File Format Full-Text
24376 43510 2010 9 PDF Available
Title
Efficient production of extracellular proteins with Escherichia coli by means of optimized coexpression of bacteriocin release proteins
Abstract

Aiming to facilitate the accessibility of recombinant proteins produced with Escherichia coli, extracellular expression may be achieved by means of bacteriocin release protein (BRP) coexpression. Different types of BRPs were tested in order to optimize protein secretion into the culture medium. Those included the well-studied BRPs of the Colicin E1 and Cloacin DF13 bacteriocins and variants thereof. BRP expression was stringently controlled by means of the arabinose inducible PBAD promoter, which accounts for a broad-range adjustment of expression strength. Using appropriate arabinose concentrations, a concentration range was determined, that allowed efficient secretion of the model proteins alkaline phosphatase and β-lactamase, with 90–95% of the proteins released into the culture medium. Kinetic investigations into BRP expression and protein secretion revealed a rapid increase of extracellular protein concentration within 5–10 min past induction. Alternatively to fine-tuned BRP expression during cultivation, protein production and secretion could be decoupled by establishment of appropriate induction strategies and up to 90% of alkaline phosphatase was released into the culture medium within 3 h after reaching maximum biomasss concentrations. Both, fine-tuned and growth decoupled BRP expression accounted for extracellular alkaline phosphatase concentrations of roughly 500 mg l−1 of culture broth and selectivities of 50 mg of this enzyme per gram of cell dry mass, respectively.

Keywords
Escherichia coli; Extracellular production; Recombinant protein expression; Secretion; Bacteriocin release protein; Promoter
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Efficient production of extracellular proteins with Escherichia coli by means of optimized coexpression of bacteriocin release proteins
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 145, Issue 4, 15 February 2010, Pages 350–358
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