Characterization of 16-kDa major allergen with α-amylase inhibitor domain in tartary buckwheat seeds. (February 2018)

• Record Type: Journal Article
• Authors: Zheng, Bei; Zhang, Haina; Wang, Lei; Guo, Yanfei; Chen, Peng
• Is Part Of: Molecular immunology. Volume 94(2018:Feb.)
• Page Start: 121
• Page End: 130
• Publication Date: 2018-02
• Subjects: 16-kDa allergen -- Tartary buckwheat seeds -- IgE binding activity -- Pepsin resistance -- Thermal stability; Immunochemistry -- Periodicals; Molecular biology -- Periodicals; Immunochemistry -- Periodicals; Allergy and Immunology -- Periodicals; Molecular Biology -- Periodicals; Immunochimie -- Périodiques; Biologie moléculaire -- Périodiques; Immunochemistry; Molecular biology; Periodicals; Electronic journals; 571.96
• DOI: 10.1016/j.molimm.2017.12.024
• Languages: English
• ISSNs: 0161-5890

Abstract

Highlights: Established the separation and identification methods of 16-kDa allergen from tartary buckwheat seeds. The recombinant Fag t 2 was expressed in Pichia pastori and purified by immobilized metal affinity chromatography. Fag t 2 was a major allergen with the activity of IgE binding and pepsin resistance, along with the thermal stability. Fag t 2 contained an AAI domain near the end of C-terminal. Abstract: Tartary buckwheat ( Fagopyrum tataricum, TB) is an important functional food containing proteins with balanced amino acid composition and more flavonoids than common buckwheat ( Fagopyrum esculentum, CB). Buckwheat contains highly potent allergens that trigger an allergic reaction via an IgE mediated response. In this work, the full-length cDNA encoding Fag t 2 from tartary buckwheat seeds was cloned by screening the cDNA library of seed-filling period. The recombinant Fag t 2 (rFag t 2) expressed in Pichia pastoris SMD1168H was purified by purified by immobilized metal affinity chromatography. It demonstrated that Fag t 2 was a major allergen in tartary buckwheat with the activity of IgE binding and pepsin resistance, along with the thermal stability. The identification of natural Fag t 2 (n Fag t 2) confirmed that the Fag t 2 protein belongs to the 2S albumin family, only existing in embryo. Most interesting, we discovered that Fag t 2 had a α-amylase inhibitor domain near the end of C-terminal. The possible activity of α-amylase inhibitor of Fag t 2 will be detected in subsequent studies.