Overexpression and Characterization of 10 kD Allergenic Protein in Tartary Buckwheat

doi:10.7506/spkx1002-6630-201403026

Abstract

Abstract:

The 10 kD allergen gene (TBAP10) of tartary buckwheat was obtained from the full-length cDNA library during
the seed-filling period, and a prokaryotic recombinant expression vector pET47b inserted with TBAP10 gene (pET47b-
TBAP10) was constructed. The target protein TBAP10 was expressed in E. coli BL21 Star (DE3) as an inclusion body
form. After re-folding and purification by cobalt ion chelating affinity chromatography of the isolated inclusion body,
allergenic activity, thermal stability and stability in simulated gastrointestinal environment of the purified TBAP10 protein
were analyzed. Western blotting revealed the immunogenic cross-reactivity between 16 kD buckwheat allergic protein and Fag t2.
Competitive ELISA demonstrated that the recombinant protein had a binding capability to IgE from buckwheat-allergic
patients. TBAP10 had strong thermal stability and could withstand boiling water bath for 15 min. Simulated gastrointestinal
digestion experiments indicated that the recombinant protein had strong tolerance to pepsin, while it could be totally
degraded after trypsin digestion.

Key words: overexpression, allergenic protein, immunological properties, simulated gastrointestinal digestion

CHEN Peng, FENG Xue, WANG Lei, DENG Dan-dan, LI Xue-jun. Overexpression and Characterization of 10 kD Allergenic Protein in Tartary Buckwheat[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201403026.

[1]	LIU Fangwei, ZHANG Shanshan, CHEN Suming, NIE Shaoping, HU Jielun, XIE Mingyong. Gastrointestinal Digestion and Fermentation Characteristics in Vitro of Breads Incorporated with Three Different Polysaccharides [J]. FOOD SCIENCE, 2021, 42(3): 143-149.
[2]	WEI Guangqiang, SHI Yanan, WAN Changjiang, HUANG Aixiang. Characterization of Peptides from Simulated Gastrointestinal Digestion of Ethnic Dairy Products in Yunnan, China [J]. FOOD SCIENCE, 2021, 42(1): 208-214.
[3]	MA Sitong, LIU Jingbo, ZHANG Ting, WANG Ying, SUN Huiyan, WEI Yiheng, WANG Hanying, LEI Honghui, LIU Boqun. Differences in Antioxidant Activity and Sequence Analysis of Egg White Peptides Derived from Simulated Gastrointestinal Digestion and Alkaline Protease Treatment [J]. FOOD SCIENCE, 2020, 41(21): 113-120.
[4]	ZHAO Lan, LIU Shimeng, QIN Hanxiong, WANG Yanan, FAN Zhanqing, MIN Weihong. Metabolic Engineering of Corynebacterium glutamicum for the Production of Methionine [J]. FOOD SCIENCE, 2020, 41(18): 98-104.
[5]	QIAN Jingjing, ZHENG Lin, ZHAO Mouming. Changing Pattern of Dipeptidyl Peptidase IV (DPP-IV) Inhibitory Activity during Casein Digestion in Vitro and Underlying Mechanism [J]. FOOD SCIENCE, 2020, 41(15): 186-193.
[6]	LIU Lei, SHE Xiao, QIAN Yang, LI Yalin, NIE Rong, RAO Yu. Effect of tuf Homogenous Overexpression on Physiological Behaviors of Lactobacillus plantarum LR-1 [J]. FOOD SCIENCE, 2019, 40(8): 48-55.
[7]	ZOU Rui, ZHANG Lingjing, ZHONG Chan, WENG Ling, LIN Liyun, LI Yujin, LIU Guangming, CAO Minjie. Characterization and Simulated Gastrointestinal Digestion of Paramyosin from Perna viridis [J]. FOOD SCIENCE, 2019, 40(6): 1-8.
[8]	CHEN Ximiao, LI Meiying, XU Qiuli, XIONG Chuxin, SUN Yuanming. Changes in Polyphenol Contents and Antioxidant Activity in Hawthorn (Crataegus pinnatifida Bunge) during Simulated Gastrointestinal Digestion [J]. FOOD SCIENCE, 2019, 40(5): 31-37.
[9]	ZHANG Chi, LI Chunyi, WANG Qiming, TANG Yuwan, ZHAO Jichun, LI Fuhua, MING Jian. Interaction between Plant Polyphenols and Peanut Allergenic Proteins and Mechanism for Allergenicity Reduction: A Review [J]. FOOD SCIENCE, 2019, 40(23): 313-318.
[10]	TONG Xiaohong, WANG Huan, LIU Baohua, ZHANG Qiaozhi, LI Hong, TIAN Ran, ZHAO Jinhan, QI Baokun, LI Yang, JIANG Lianzhou. Changes in Antioxidant Activity of Soybean Protein Hydrolysates from Enzyme-Assisted Aqueous Extraction of Oil under Simulated Gastrointestinal Digestion [J]. FOOD SCIENCE, 2019, 40(15): 50-56.
[11]	HE Jilong, LI Yingzi, HAN Shibao, MAN De’en, GUO Maihai, ZHAN Junjie, ZHANG Chenglin. Effect of Constitutively Overexpressed ido Gene on 4-Hydroxyisoleucine Production and Optimization of the Fermentation Medium by Response Surface Methodology [J]. FOOD SCIENCE, 2019, 40(14): 91-98.
[12]	LIN Lin, WANG Changlu, LI Zhenjing, CHEN Mianhua, WU Shufen, REN Zhiyuan. Effect of mok E Overexpression on Monacolin K Production and Morphology of Mycelia and Spores in Monascus [J]. FOOD SCIENCE, 2018, 39(8): 45-49.
[13]	LU Jun, DUN Huiyu, XIANG Xiaozhe, FU Chunya, MO Kaidi, ZENG Xian. Effect of in Vitro Simulated Gastrointestinal Digestion on Bioactive Components and Antioxidant Activities of Six Kinds of Black Foods [J]. FOOD SCIENCE, 2018, 39(5): 47-56.
[14]	QU Xiuwei, LIU Lu, WANG Haixia, LI Xiaodong, ZHANG Xiuxiu, CHEN Ping, SHEN Qi. Antioxidant Capacity of Probiotic Cheese after Simulated Gastrointestinal Digestion [J]. FOOD SCIENCE, 2018, 39(3): 90-96.
[15]	QIAN Hui, LU Zhaoxin, ZHANG Chong, BIE Xiaomei, ZHAO Haizhen, Lü Fengxia. Molecular Dynamics Simulation of Thermal Stability Improvement of Lipoxygenase from Anabaena sp. PCC 7160 [J]. FOOD SCIENCE, 2017, 38(2): 1-6.

Overexpression and Characterization of 10 kD Allergenic Protein in Tartary Buckwheat

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments