凡纳滨对虾主要过敏原鉴定及酶法消减技术的研究

以凡纳滨对虾为研究对象，用免疫印迹法检测其主要过敏原蛋白组分；用胰蛋白酶、木瓜蛋白酶和菠萝蛋白酶对凡纳滨对虾水溶性蛋白水解，消减过敏原。并且通过间接酶联免疫吸附实验和致敏豚鼠全身免疫实验对过敏原消减情况进行检测，确定酶法消减过敏原条件。结果表明，凡纳滨对虾的主要过敏原为99、33、19kD 以及14kD 的蛋白质组分。胰蛋白酶最佳水解条件为：pH8.0，酶与底物比1:100(m/m，下同)，45℃，底物质量浓度3g/100mL，水解时间3h；木瓜蛋白酶最佳水解条件为pH6.5，酶与底物比1:100，60℃，底物质量浓度3g/100mL，水解时间3h；菠萝蛋白酶最佳水解条件：pH7.5，酶与底物比1:100，55℃，底物质量浓度5g/100mL，水解时间3h。并且胰蛋白酶水解产物和木瓜蛋白酶水解产物对致敏豚鼠的全身免疫实验安全性很好。

Identification and Enzymatic Elimination of Major Allergens in Litopenaeus vannamei

The major allergenic protein components in Litopenaeus vannamei were determined by Western blotting. Either trypsin, papain or bromelain was used to hydrolyze water-soluble proteins from Litopenaeus vannamei for the elimination of allergens. ELISA and general immunity experiments in allergic guinea pigs were used to evaluate the elimination of allergens. The optimum hydrolysis conditions for achieving a maximum elimination of allergens were also determined. The results showed that the major allergens in Litopenaeus vannamei were found to have molecular weights of 99, 33, 19 kD and 14 kD, respectively. The optimal conditions for trypsin hydrolysis were as follows: pH 8.0, hydrolysis temperature 45 ℃, enzyme-to-substrate ratio 1: 100, protein concentration 3 g/100 mL and hydrolysis time 3 h; those for papain digestion were pH 6.5, hydrolysis temperature 60 ℃, enzyme-to-substrate ratio 1:100, protein concentration 3 g/100 mL and hydrolysis time of 3 h; and pH of 7.5, hydrolysis temperature of 55 ℃, enzyme-substrate ratio of 1:100, protein concentration of 5 g/100 mL and hydrolysis time of 3 h were found optimum for bromelain hydrolysis. The hydrolysates derived from trypsin and papain hydrolysis were safer for allergic guinea pigs.