Purification and characterization of aβ-glucosidase (linamarase) from the haemolymph ofZygaena trifolii Esper, 1783 (Insecta, Lepidoptera)
Purification and characterization of aβ-glucosidase (linamarase) from the haemolymph ofZygaena trifolii Esper, 1783 (Insecta, Lepidoptera)
复制标题
从 Zygaena trifolii Esper, 1783（昆虫纲，鳞翅目）血淋巴中纯化和鉴定 α-葡萄糖苷酶（亚麻酶）
DOI：
10.1007/bf01974565
复制
发表时间：
1989
期刊：
影响因子：
--
通讯作者：
A. Nahrstedt
中科院分区：
文献类型：
--
作者：
S. Franzl;I. Ackermann;A. Nahrstedt
从三叶接合藻（Zygaenatrifolii，ESPER，1783）幼虫血淋巴中纯化了一种β-葡萄糖苷酶（linamarase），纯化倍数为52倍，回收率为27%。发现最终的酶制剂在圆盘聚丙烯酰胺凝胶电泳和SDS-聚丙烯酰胺凝胶电泳上几乎是均一的。该酶的分子量被确定为约130 kDa;它由两个约66 kDa的亚基组成。该酶对亚麻苦苷的最适pH为4.5 ~ 5，对对硝基苯基-β-D-葡萄糖苷的最适pH为3.5 ~ 6.5，最适温度为40°C。β-葡萄糖苷酶对内源底物亚麻苦苷和百脉根苷具有高度的专一性。在其他天然和人工底物中，只有Prunasin和对硝基苯基-β-D-葡萄糖苷被该酶水解，而Linustatin、水杨苷、纤维二糖和海藻糖不被该酶水解。β-葡糖基哌啶强烈抑制该酶。
A β-glucosidase (linamarase) was purified 52-fold with a recovery of 27% from the haemolymph of the larvae ofZygaena trifolii, ESPER, 1783 (Lepidoptera, Zygaenidae). The final enzyme preparation was found to be nearly homogeneous on both disc polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis. The molecular weight of the enzyme was determined to be about 130 kDa; it consisted of two subunits of about 66 kDa. The enzyme showed an optimum between pH 4.5 and 5 with linamarin and a broad optimum between pH 3.5 and 6.5 for p-nitrophenyl-β-D-glucoside; the temperature optimum was 40°C. The β-glucosidase showed a high specificity for its endogenous substrates linamarin and lotaustralin. Among the other natural and artificial substrates tested, only prunasin and p-nitrophenyl-β-D-glucoside were hydrolyzed by the enzyme, whereas linustatin, salicin, cellobiose and trehalose were not. The enzyme is strongly inhibited by β-glucosylpiperidine.