Document Type

Article

Research Focus Area

Biorenewables

Publication Date

10-18-2006

Journal or Book Title

Journal of Agricultural and Food Chemistry

Volume

54

Issue

21

First Page

8086

Last Page

8092

DOI

10.1021/jf061921h

Abstract

Several approaches were examined for extracting the relatively hydrophobic protein recombinant dog gastric lipase (rDGL) expressed in the endosperm of transgenic corn seed. The first approach used minimal processing of the seed before extraction (i.e. simple grinding of whole seed) followed by selective extraction to eliminate 72% of contaminant proteins without compromising rDGL recovery from the meal of whole grain. The second approach added defatting of the whole grain meal to reduce the amount of detergent in the subsequent step for extracting rDGL. The third approach incorporated dry-milling of the corn to recover an endosperm rich fraction, followed by extraction of this fraction. The dry milling strategy was most effective, resulting in recovery of 35 U rDGL/g of corn seed (50 U/g of endosperm) with a specific activity of 9 U/mg compared to 22 U and 3 U/mg for the first strategy and 36 U and 3.7 U/mg for the second. The reductions in host protein contamination and lower detergent levels of the endosperm route should simplify downstream purification steps.

Comments

Reprinted with permission from Journal of Agricultural and Food Chemistry 54 (2006): 8086–8092, doi:10.1021/jf061921h. Copyright 2006 American Chemical Society.

Copyright Owner

American Chemical Society

Language

en

File Format

application/pdf

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