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Process optimization for recombinant protein production in the psychrophilic bacterium Pseudoalteromonas haloplanktis

Paper ID Volume ID Publish Year Pages File Format Full-Text
34883 45056 2011 7 PDF Available
Title
Process optimization for recombinant protein production in the psychrophilic bacterium Pseudoalteromonas haloplanktis
Abstract

A new regulated expression system has been developed for recombinant protein production in Pseudoalteromonas haloplanktis, strongly induced by the presence of l-malate in culture medium. In optimized induction conditions, high yields of soluble recombinant model proteins were obtained in small scale expression experiments. However, performances of the new expression system are still not satisfactory for large scale recombinant protein production. To improve the psychrophilic expression system, the influence of medium composition and cultivation operational strategies on biomass concentration, growth rate and protein production have been investigated in this study. A new defined medium, containing branched amino acids (L, I, V) as carbon sources, was developed resulting in over sevenfold increase of reporter enzyme production and threefold of biomass yield with respect to the previously optimized conditions. The new synthetic medium was also tested for the production of a recombinant antibody fragment. In batch cultivation, up to 0.9 mg g dry cell weight −1 of soluble 3H6 Fab were recovered in bacterial periplasm. Moreover, the new optimized medium was tested for P. haloplanktis chemostat cultivation. A continuous process for 3H6 Fab production was developed leading to a volumetric productivity of about 0.2 mg L−1 h−1 of soluble and periplasmic recombinant antibody.

Keywords
Psychrophiles; Recombinant protein production; Antibody fragment; 3H6 Fab; Synthetic medium
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Process optimization for recombinant protein production in the psychrophilic bacterium Pseudoalteromonas haloplanktis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 46, Issue 4, April 2011, Pages 953–959
Authors
, , , , , ,
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