Document Type

Article

Research Focus Area

Biorenewables

Publication Date

2-11-2009

Journal or Book Title

Journal of Agricultural and Food Chemistry

Volume

57

Issue

3

First Page

880

Last Page

887

DOI

10.1021/jf8026205

Abstract

This paper demonstrates that a fibrous, repetitive amino acid sequence collagen-related protein, a 44-kDa fragment of human collagen I α 1 (CIα1), was expressed in corn grain molecularly equivalent to that produced in recombinant yeast. The recombinant CIα1 was extracted and purified from early generation plants having low levels of recombinant protein accumulation. It was selectively extracted at low pH and purified by ion exchange and gel filtration chromatography, resulting in a 44-kDa CIα1 with >70% purity and 60% recovery. The N-terminal sequence, amino acid composition, and immunoreactivity closely matched those of an analogous 44-kDa CIα1 fragment produced by the yeast Pichia. The corn-derived 44-kDa CIα1 had an intact protein mass of 44088 Da, which is within 0.2% of the mass calculated from the expected sequence. Tandem mass spectrometry confirmed the primary sequence with 78% coverage. The amino acid composition analysis indicated a low level of prolyl hydroxylation. Glycoprotein staining revealed no glycosylation.

Comments

Reprinted with permission from Journal of Agricultural and Food Chemistry 5 (2009): 880–887, doi:10.1021/jf8026205. Copyright 2009 American Chemical Society.

Copyright Owner

American Chemical Society

Language

en

File Format

application/pdf

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