Identify The Major Allergens And Research On Technology Of Elimination To Major Allergens Of Litopenaeus Vannamei

It was studied that the shrimp allergen protein components,and Litopenaeus vannamei,Fenneropenaeus Chinensis,Metapenaeus ensis and mantis shrimp were used as subjects in the experiment.Frist,to analysis the four kinds of shrimp allergy components by SDS-PAGE,and then identified the major allergens by Western blotting.The result showed that Litopenaeus vannamei major allergens are mainly 99 kDa,33kDa,19 kDa and 14kDa;Fenneropenaeus Chinensis major allergens are mainly 39 kDa,33kDa and 28kDa;Metapenaeus ensis major allergens are mainly 33 kDa and 24kDa;mantis shrimp major allergens are mainly 56 kDa and48kDa.To hydrolyze the water-soluble protein components by using enzyme(Trypsin,Papain and Bromelain)and reduce allergen.To detect the extent of reducing by ELISA,then determine the best conditions of three enzyme(Trypsin,Papain and Bromelain)reduction allergen by the OD value of 492 nm.The optimize conditions of the Litopenaeus vannamei protein hydrolyzed by trypsin as follows: pH value is 8.0,the ratio of enzyme to substrate is 1:100,hydrolysis temperature is 45℃,protein concentration is 3%,hydrolysis time is 3h;the optimize conditions of the Litopenaeus vannamei protein hydrolyzed by papain as follows: pH value is 6.5,the ratio of enzyme to substrate is 1:100,hydrolysis temperature is 60℃,protein concentration is 3%,hydrolysis time is 3h;the optimize conditions of the Litopenaeus vannamei protein hydrolyzed by bromelain as follows: pH value is7.5,the ratio of enzyme to substrate rate is 1:100,hydrolysis temperature is 55℃,protein concentration is 5%,hydrolysis time is 3h.Using egg albumin as the positive control and the PBS solution as the negative control to observe general allergic reaction of shrimp protein and the isolated ileal smooth muscle contraction(Schultz-Dale reaction)of guinea pig,to establish an allergy animal experiment model.Results showed that the general allergic reaction in each guinea pig were positive after stimulated by egg albumin at concentration varying from 0.01 to 2 g/100 m L.The general allergic reaction of protein of Litopenaeus vannamei were similar with the positive control at the similar concentration of 0.01 to 1 g/100 mL,and the Schultz-Dale reaction results were also similar with those of systemic allergic reaction,so an animal model for shrimp protein sensitivity test has been successfully established.Using general immunity experiments in allergic pig and allergic contraction experiments of isolated ileum to evaluate the allergenicity of the three hydrolysates(proteases such as: Trypsin,papain and bromelain).The results showed an obvious attenuation of general allergic reaction of the hydrolysates from Litopenaeus vannamei protein digested by the three proteases.Meanwhile,a reduced allergic contraction reaction of isolated ileum smooth muscle was observed in hydrolyzed Litopenaeus vannamei protein.It was studied that the allergen of Litopenaeus vannamei which was hydrolyzed by Trypsin and Papain.To detect the extent of reducing by ELISA then determine the best conditions of enzymatic reduction allergen by the OD value of 492 nm.The results showed that the optimize conditions of the Litopenaeus vannamei hydrolyzed by trypsin as follows: pH value is 8.0,the ratio of enzyme to substrate is 1:100,hydrolysis temperature is 45℃,protein concentration is 5%,hydrolysis time is 3h,the OD value of hydrolysis was 0.085;the optimize conditions of the Litopenaeus vannamei hydrolyzed by papain as follows: pH value is 6.5,the ratio of enzyme to substrate is 1:100,hydrolysis temperature is 60℃,protein concentration is 5%,hydrolysis time is 3h,the OD value of hydrolysis was 0.049.The best protein of hydrolyzing Litopenaeus vannamei is the Papain.