胰蛋白酶、SDS对三疣梭子蟹酚氧化酶活力的影响

Effects of Trypsin and SDS on the activity of Phenoloxidase from Portunus trituberculatus

本文研究了胰蛋白酶、SDS对三疣梭子蟹血浆血细胞中酚氧化酶活性的影响,分别测定了血浆、血细胞裂解溶液中酚氧化酶活性;SDS和胰蛋白酶分别刺激血浆和血细胞裂解液后酚氧化酶活力变化,分析了酚氧化酶原系统在三疣梭子蟹血淋巴的存在位置以及其应答机制.结果表明,三疣梭子蟹血细胞裂解液中的酚氧化酶活力(9.93U±1.03U)高于血浆中酚氧化酶活力(0.80U±0.04U);胰蛋白酶对三疣梭子蟹血浆和血细胞裂解液中的酚氧化酶活力激活作用不明显,分别为0.91U±0.07U和9.06U±3.65U;SDS对三疣梭子蟹血浆中酚氧化酶活力有激活作用,为0.96U±0.03U,对血细胞裂解产物中酚氧化酶活力的有显著激活作用,为71.12U±14.25U.三疣梭子蟹的酚氧化酶原系统主要存在于血细胞中,既可以通过多级酶联免疫反应将酚氧化酶原激活成酚氧化酶参与免疫反应,也可以通过与酚氧化酶原结合,改变其蛋白构象,激活酚氧化酶.胰蛋白酶通过前一种方式激活酚氧化酶原系统,而SDS通过后一种方式激活,SDS对酚氧化酶原的激活速率远高于胰蛋白酶.

In this research,the effect of Trypsin and SDS on Portunus trituberculats was analyzed. Phenoloxdase（ PO） of plasma and hemocytely supernatan（ HLS） were measured as well as the Kinetic curve of PO activity of Trypsin and SDS conducted on plasma and HLS; analysis the prophenol- oxidase activated system（ Pro PO- AS） existing position and immune mechanism of Portunus trituberculatus. The results showed that PO activety of HLS（ 9. 93 U ± 1. 03 U） is higher than plams（ 0. 80 U ± 0. 04 U）; the effect of Trypsin conducted on plasma and HLS was not significant were： 0. 91 U ± 0. 07 U and 9. 06 U ± 3. 65 U; the effect of SDS conducted on plasma and HLS was obvious were 0. 96 U ± 0. 03 U and 71. 12 U ±14. 25 U. Pro PO- AS of Portunus trituberculatusexist in hemocytely,it participate the immune response through different paths： Trypsin through the multi- stage enzyme- linked immunosorbent reaction to oxidize Pro PO to PO while SDS active PO by change the structure of Pro PO. The efficiency of SDS active Pro PO- AS is significanly higher than Trypsin.