褶皱臂尾轮虫（轮虫纲）中的糖苷酶

Glycosidases in Brachionus plicatilis (Rotifera).

Kühle K, Kleinow W

Zoologisches Institut der Universität zu Köln, West Germany.

Comp Biochem Physiol B. 1990;95(2):393-402. doi: 10.1016/0305-0491(90)90093-9.

Abstract

Tests for glycosidases were performed in homogenates of Brachionus plicatilis. 2. Hydrolytic activity was detected with the following substrates: (a) with synthetic substrates (NP = 4-nitrophenyl): NP-alpha- and NP-beta-D-glucopyranoside, NP-alpha- and NP-beta-D-galactopyranoside, NP-N-acetyl-beta-D-glucosaminide, NP-N-acetyl-beta-D-galactosaminide, NP-alpha- and NP-beta-D-mannopyranoside and NP-alpha-L-fucopyranoside; (b) with disaccharides: sucrose, maltose, trehalose, isomaltose, cellobiose, gentiobiose and lactose; (c) with polysaccharides: laminarine, carboxymethyl-cellulose, avicel, Micrococcus luteus (for lysozyme) and 4-nitrophenyl-alpha-D-maltoheptaoside (for amylase). 3. The pH dependence of the glycosidase activities was determined. 4. The distribution of enzyme activities within fractions from the homogenate was studied in order to localize them within the cell. 5. Proteins from Brachionus homogenate were separated by SDS-gel electrophoresis and the positions of the following glycosidase activities were detected by assays performed on the gels (estimated molecular weights in parentheses): alpha-glucosidase (250,000); beta-glucosidase (200,000); beta-galactosidase (70,000); N-acetyl-beta-glucosaminidase (60,000).

摘要

在褶皱臂尾轮虫的匀浆中进行糖苷酶测试。2. 用以下底物检测水解活性：(a) 合成底物（NP = 4-硝基苯基）：NP-α-和NP-β-D-吡喃葡萄糖苷、NP-α-和NP-β-D-吡喃半乳糖苷、NP-N-乙酰-β-D-氨基葡萄糖苷、NP-N-乙酰-β-D-半乳糖胺、NP-α-和NP-β-D-甘露糖苷以及NP-α-L-岩藻糖苷；(b) 二糖：蔗糖、麦芽糖、海藻糖、异麦芽糖、纤维二糖、龙胆二糖和乳糖；(c) 多糖：海带多糖、羧甲基纤维素、微晶纤维素、藤黄微球菌（用于溶菌酶）和4-硝基苯基-α-D-麦芽庚糖苷（用于淀粉酶）。3. 测定糖苷酶活性的pH依赖性。4. 研究匀浆各组分中酶活性的分布，以便将它们定位在细胞内。5. 通过SDS凝胶电泳分离褶皱臂尾轮虫匀浆中的蛋白质，并通过对凝胶进行测定来检测以下糖苷酶活性的位置（括号内为估计的分子量）：α-葡萄糖苷酶（250,000）；β-葡萄糖苷酶（200,000）；β-半乳糖苷酶（70,000）；N-乙酰-β-氨基葡萄糖苷酶（60,000）。