FOOD SCIENCE ›› 2010, Vol. 31 ›› Issue (13): 169-173.doi: 10.7506/spkx1002-6630-201013039

• Bioengineering • Previous Articles     Next Articles

Immunological Characterization and Mutational Analysis of Allergen TBa in Tartary Buckwheat

REN Xiao-xia1，ZHANG Xin2，CUI Xiao-dong1，LI Yu-ying1，WANG Zhuan-hua1,*   

  1. 1. Key Laboratory of Chemical Biology and Molecular Engineering, Ministry of Education, Institute of Biotechnology,
    Shanxi University, Taiyuan 030006, China；2. Research Center for Environmental Science and Engineering, Shanxi University,
    Taiyuan 030006, China
  • Received:2009-10-23 Revised:2010-04-21 Online:2010-07-01 Published:2010-12-29
  • Contact: WANG Zhuan-hua E-mail:zhwang@sxu.edu.cn

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Abstract:

Buckwheat not only contains abundant nutriments such as flavoniods, but also has allergenic proteins that cause hypersensitive reactions. A natural allergenic protein with molecular weight of 24 kD named TBa (tartary buckwheat allergen A) has been obtained after separation and purification in our previous studies. According to its biological information, 5 mutants including L39R, L42R, L47R, V52R and L54R were constructed through site-directed mutagenesis and expressed in Escherichia coli BL21. The expressed products were purified by Ni2+ affinity chromatography. Purified recombinant proteins were examined using ELISA and dot blot and mutants L42R, L47R and L54R presented a weaker ability to bind tartary buckwheat allergic patients serum IgE than original E1, which suggests that Leu42, Leu47 and Leu54 might play a significant role in TBa allergenity.

Key words: tartary buckwheat, allergen, site-directed mutagenesis, immunological activity

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