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Journal of Agricultural and Food Chemistry 2014-Dec

Expression, purification, and characterization of almond (Prunus dulcis) allergen Pru du 4.

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Povezava se shrani v odložišče
Yuzhu Zhang
Wen-Xian Du
Cécile Fregevu
Mahendra H Kothary
Leslie Harden
Tara H McHugh

Ključne besede

Povzetek

Biochemical characterizations of food allergens are required for understanding the allergenicity of food allergens. Such studies require a relatively large amount of highly purified allergens. The level of Pru du 4 in almond is low, and its expression in a soluble form in Escherichia coli required an expression tag. An MBP tag was used to enhance its expression and solubility. Sumo was used for the first time as a peptidase recognition site. The expression tag was removed with a sumo protease, and the resulting wild-type Pru du 4 was purified chromatographically. The stability of the allergen was investigated with chemical denaturation. The Gibbs free energy of Pru du 4 folding-unfolding transition was determined to be 5.4 ± 0.7 kcal/mol.

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