Food proteins from yeast-based precision fermentation: Simple purification of recombinant β-lactoglobulin using polyphosphate.

Autor: Hoppenreijs LJG; Laboratory of Food Process Engineering, Wageningen University & Research, Bornse Weilanden 9, 6708 WG Wageningen, the Netherlands., Annibal A; Formo Bio GmbH, Weismüllerstraße 50, 60314 Frankfurt am Main, Germany., Vreeke GJC; Laboratory of Food Chemistry, Wageningen University & Research, Bornse Weilanden 9, 6708 WG Wageningen, the Netherlands., Boom RM; Laboratory of Food Process Engineering, Wageningen University & Research, Bornse Weilanden 9, 6708 WG Wageningen, the Netherlands., Keppler JK; Laboratory of Food Process Engineering, Wageningen University & Research, Bornse Weilanden 9, 6708 WG Wageningen, the Netherlands. Electronic address: Julia.keppler@wur.nl.
Jazyk: angličtina
Zdroj: Food research international (Ottawa, Ont.) [Food Res Int] 2024 Jan; Vol. 176, pp. 113801. Date of Electronic Publication: 2023 Dec 04.
DOI: 10.1016/j.foodres.2023.113801
Abstrakt: Proteins produced through precision fermentation are often purified through chromatographic methods. Faster and more cost-effective purification methods are desired for food application. Here, we present a simple method for purification of protein produced from yeast, using β-lactoglobulin secreted from Pichia pastoris as an example. The food-grade salt hexametaphosphate (HMP) was used to precipitate the protein at acidic pH, while the impurities (extracellular polysaccharides; mainly mannan) remained soluble. After re-solubilization of the protein-HMP complex by neutralization, excess HMP was selectively precipitated using calcium chloride. The protein content of the crude sample increased from 26 to 72 wt% (comparable to purification with anion exchange chromatography), containing only residual extracellular polysaccharides (9 wt%) and HMP (1 wt%). The established method had no significant impact on the structural and functional properties (i.e., ability to form emulsions) of the protein. The presented method shows potential for cost-effective purification of recombinant proteins produced through yeast-based expression systems.
Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Andrea Annibal is employed by a company that develops alternative dairy products using precision fermentation.
(Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
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