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Secreted production of self-assembling peptides in Pichia pastoris by fusion to an artificial highly hydrophilic protein

Paper ID Volume ID Publish Year Pages File Format Full-Text
24514 43521 2010 8 PDF Available
Title
Secreted production of self-assembling peptides in Pichia pastoris by fusion to an artificial highly hydrophilic protein
Abstract

The undecapeptides CH3CO-Gln-Gln-Arg-Phe-Gln-Trp-Gln-Phe-Glu-Gln-Gln-NH2 (P11-2) and CH3CO-Gln-Gln-Orn-Phe-Orn-Trp-Orn-Phe-Orn-Gln-Gln-NH2 (P11-14) have unique self-assembly characteristics and broad application potential. Originally, these peptides were produced by chemical synthesis, which is costly and difficult to scale up to industrial levels in an economically feasible way. This article describes the efficient secreted production of these peptides (with free termini and ornithines replaced with lysines) in the methylotrophic yeast Pichia pastoris. The peptides were produced as enterokinase-cleavable fusions to the C-terminus of an artificial Solubility-Enhancing Protein (SEP). In vitro, the fused highly hydrophilic SEP proved to prevent self-assembly of the peptides. The SEP domain also facilitates product detection and allows convenient separation of the fusion protein from the broth by simple salt precipitation. After cleavage of the purified fusion protein with enterokinase, the free undecapeptides were obtained and P11-2 spontaneously assembled into a self-supporting gel, as intended. The properties of the SEP carrier could be advantageous for the production of other peptides.

Keywords
Enterokinase; Fusion protein; Pichia pastoris; Protein secretion; Self-assembling peptide; Self-supporting gel
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Secreted production of self-assembling peptides in Pichia pastoris by fusion to an artificial highly hydrophilic protein
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 146, Issues 1–2, March 2010, Pages 66–73
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