Isolation, purification and identification of sarcoplasmic calcium-binding protein from oyster (Crassostrea gigas) allergen
CHENG Qingli, LI Guoming, ZHAO Xiaohan, FENG Xiaowen, LU Zhihao, GU Ruizeng, LU Jun, LIU Wenying*
(Beijing Engineering Research Center of Protein and Functional Peptides, China National Research Institute of Food and Fermentation Industries, Beijing 100015, China)
Abstract: The aim of this study was to identify the single protein extracted from Crassostrea gigas muscle sarcoplasm by mass spectrometry and analyze the molecular weight consistency of the induced recombinant protein and the natural protein identified by western blot(WB). The consistency of the two protein on the two level structure was analyzed by cireular dichroism(CD) spectrum, and the sarcoplasmic calcium-binding protein (SCP) glycoprotein type of oyster was identified. The content of polysaccharide was also determined. The results revealed that 20 kDa protein was purified from oyster crude extract and confirmed as SCP by LC-MS / MS,the bands of natural SCP and recombinant SCP were 20 kDa and 22 kDa respectively,which including 13.5%, 18.9% and 14.5%, 18.2% Helix and Beta-Turn , respectively. SCP protein contained 7.89% O-glycosidic glycopolysaccharide. The molecular weight of recombinant SCP was slightly different from that of natural SCP, but their spatial structures were almost the same. It is a new potential allergen in Crassostrea gigas, which lays an important foundation for the study of allergenicity and sensitization mechanism of recombinant SCP instead of natural SCP in the future.
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2021, Vol. 47 
