Purification and characterization of extracellular alpha-amylase and glucoamylase from the yeast Candida antarctica CBS 6678.
Purification and characterization of extracellular alpha-amylase and glucoamylase from the yeast Candida antarctica CBS 6678.
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南极假丝酵母 CBS 6678 酵母细胞外 α-淀粉酶和葡糖淀粉酶的纯化和表征。
DOI：
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
H. Verachtert
中科院分区：
文献类型：
--
作者：
R. Mot;H. Verachtert
经硫酸鱼精蛋白处理、硫酸铵沉淀、凝胶过滤(Sephadex G-75 SF，Ultroguel ACA 54)、DEAE-Sephacel柱层析、羟基磷灰石柱层析和阿卡波糖-AH-Sepharose4B亲和层析，从培养的南极假丝酵母CBS 6678培养液中分离纯化得到一种α-淀粉酶和一种糖化淀粉酶。这两种酶都是氨基酸组成相当不同的单体糖蛋白。糖淀粉酶的表观相对分子质量、沉降系数(Szero20，w)、等电点、吸收系数(280 Nm)、pH和最适温度分别为48,500，4.7 S，10.1，1.74 cm~2 mg~(-1)，4.2和57℃；动力学分析表明，这两种酶都优先水解高分子量底物，包括一些粗淀粉。α-淀粉酶对环糊精有活性，而糖淀粉酶有去支化活性。Trestatins对α-淀粉酶(Ki<1微米)和糖淀粉酶(Ki<0.1微米)都有很强的抑制作用，比Bay e 4609(Ki<10微米)更有效。糖化淀粉酶具有选择性，并被阿卡波糖(KI<0.1微米)强烈抑制。1-脱氧野生霉素(KI<1 mM)、麦芽糖醇和氨基醇(KI<10 mM)对酶活力也有影响。与α-淀粉酶不同的是，糖化淀粉酶对原淀粉有很强的吸附作用，吸附部位与活性部位不同。
An alpha-amylase and a glucoamylase were purified to homogeneity from the culture fluid of beta-cyclodextrin-grown Candida antarctica CBS 6678 by protamine sulfate treatment, ammonium sulfate precipitation, gel filtration (Sephadex G-75 sf, Ultrogel AcA 54), DEAE-Sephacel chromatography, hydroxyapatite chromatography and affinity chromatography on acarbose--AH-Sepharose 4B. Both enzymes were monomeric glycoproteins with fairly different amino acid compositions. Their apparent relative molecular mass, sedimentation coefficient (Szero20,w), isoelectric point, absorption coefficient (280 nm), pH and temperature optima were estimated as 48,500, 4.7 S, 10.1, 1.74 cm2 mg-1, 4.2 and 57 degrees C, respectively, for glucoamylase and as 50,000, 4.9 S, 10.3, 1.53 cm2 mg-1, 4.2 and 62 degrees C, respectively, for alpha-amylase. Kinetic analyses indicated that both enzymes preferentially hydrolyzed high-molecular-mass substrates, including some raw starches. alpha-Amylase was active on cyclodextrins, whereas debranching activity was demonstrated for glucoamylase. Trestatins were potent inhibitors of both alpha-amylase (Ki less than 1 microM) and glucoamylase (Ki less than 0.1 microM), being more effective than Bay e 4609 (Ki less than 10 microM). Glucoamylase was selectivity and strongly inhibited by acarbose (Ki less than 0.1 microM). Activity of the latter enzyme was also affected by 1-deoxynojirimycin (Ki less than 1 mM), maltitol and amino alcohols (Ki less than 10 mM). Unlike alpha-amylase, glucoamylase adsorbed strongly onto raw starch, the adsorption site being non-identical with the active site.
糖化酵母 STA2 编码的胞外葡糖淀粉酶的生化和免疫学特征。
影响因子：
3.9
作者：
Modena,D;Vanoni,M;Englard,S;Marmur,J
通讯作者：
Marmur,J