{"title":"蜜蜂芽孢杆菌CBML 152中耐热α -淀粉酶的纯化及其性质研究。","authors":"S B Ghosh, A K Chandra","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>A thermostable alpha-amylase was purified from Bacillus apiarius CBML 152 by ethanol precipitation, starchamylase complex formation and by ion-exchange chromatography using DEAE-cellulose at pH 8.6, eluted with 0.2 to 0.3 M NaCl in the starting buffer (0.5 M tris-HCl, pH 8.6). The purified enzyme was homogeneous in polyacrylamide gel electrophoresis. The molecular weight of the enzyme was found to be about 65,000 dalton by gel electrophoresis in the presence of SDS. Characteristically the enzyme was an acidic protein, highly pH stable and thermostable, retaining 100% activity after being exposed to 60 degrees C for 1 hr and to pH 6.0-9.0 for 48 hr. Some divalent cations, as Ca2+, Fe2+, Mn2+ and Zn2+, stimulated the enzyme activity at different molar concentrations.</p>","PeriodicalId":75427,"journal":{"name":"Acta vitaminologica et enzymologica","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1984-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Purification and some properties of a thermostable alpha-amylase from Bacillus apiarius CBML 152.\",\"authors\":\"S B Ghosh, A K Chandra\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>A thermostable alpha-amylase was purified from Bacillus apiarius CBML 152 by ethanol precipitation, starchamylase complex formation and by ion-exchange chromatography using DEAE-cellulose at pH 8.6, eluted with 0.2 to 0.3 M NaCl in the starting buffer (0.5 M tris-HCl, pH 8.6). The purified enzyme was homogeneous in polyacrylamide gel electrophoresis. The molecular weight of the enzyme was found to be about 65,000 dalton by gel electrophoresis in the presence of SDS. Characteristically the enzyme was an acidic protein, highly pH stable and thermostable, retaining 100% activity after being exposed to 60 degrees C for 1 hr and to pH 6.0-9.0 for 48 hr. Some divalent cations, as Ca2+, Fe2+, Mn2+ and Zn2+, stimulated the enzyme activity at different molar concentrations.</p>\",\"PeriodicalId\":75427,\"journal\":{\"name\":\"Acta vitaminologica et enzymologica\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1984-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Acta vitaminologica et enzymologica\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Acta vitaminologica et enzymologica","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
通过乙醇沉淀、形成淀粉酰酶复合物和deae -纤维素离子交换层析,在pH 8.6的条件下,用0.2 ~ 0.3 M NaCl在起始缓冲液(0.5 M tris-HCl, pH 8.6)中洗脱,从apiarius芽孢杆菌CBML 152中纯化出一种热稳定型α -淀粉酶。纯化后的酶在聚丙烯酰胺凝胶电泳中均相。在SDS存在下,凝胶电泳发现酶的分子量约为65000道尔顿。该酶的特点是一种酸性蛋白质,具有高度的pH稳定性和热稳定性,在60℃下暴露1小时,在pH 6.0-9.0下暴露48小时后保持100%的活性。Ca2+、Fe2+、Mn2+和Zn2+等二价阳离子在不同摩尔浓度下均能刺激酶活性。
Purification and some properties of a thermostable alpha-amylase from Bacillus apiarius CBML 152.
A thermostable alpha-amylase was purified from Bacillus apiarius CBML 152 by ethanol precipitation, starchamylase complex formation and by ion-exchange chromatography using DEAE-cellulose at pH 8.6, eluted with 0.2 to 0.3 M NaCl in the starting buffer (0.5 M tris-HCl, pH 8.6). The purified enzyme was homogeneous in polyacrylamide gel electrophoresis. The molecular weight of the enzyme was found to be about 65,000 dalton by gel electrophoresis in the presence of SDS. Characteristically the enzyme was an acidic protein, highly pH stable and thermostable, retaining 100% activity after being exposed to 60 degrees C for 1 hr and to pH 6.0-9.0 for 48 hr. Some divalent cations, as Ca2+, Fe2+, Mn2+ and Zn2+, stimulated the enzyme activity at different molar concentrations.