• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>African Journal of Biotechnology >Purification and biochemical characterization of a Ca2+-independent, thermostable and acidophilic -amylase from Bacillus sp. RM16
【24h】

Purification and biochemical characterization of a Ca2+-independent, thermostable and acidophilic -amylase from Bacillus sp. RM16

机译：芽孢杆菌属的一种不依赖于Ca2 +的，热稳定的和嗜酸性的淀粉酶的纯化和生化特性。 RM16

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Bacillus?sp. RM16 was isolated from a hot spring in Karachi and screened for the production of α-amylase. The enzyme was obtained after 72 h cultivation of strain in Luria broth containing 1% starch (w/v). Enzyme Amy RM16 was purified to electrophoretic homogeneity by a series of sequential steps including precipitation with ammonium sulfate at 70% saturation, Q-Sepharose, Phenyl Sepharose and reversed phase chromatography. The purified enzyme is made up of a single polypeptide chain of 66 kDa as established by a combination of SDS-PAGE and zymographic analysis. In our experimental conditions, a total yield of 1.35% with specific activity of 6380U/mg was obtained providing 17 fold final purification of the enzyme. Biochemical characterization of the Amy RM16 such as optimum temperature and pH, substrate specificity and enzymatic susceptibilities towards different metal ions and inhibitors were also performed. Results of these studies revealed that, the enzyme is active at wide temperature range with optimum activity at 80°C and retained 85% of the activity for 3 h at 50°C and around 50% of remaining activity for 1 h at 80°C. The enzyme showed optimum activity at pH 5.0. On the other hand, Ca+2?and EDTA (1 to 5 mM) did not significantly affect the enzyme activity. The main substrate for the enzyme was found to be starch but it could also hydrolyze raw starch, dextrin,?g--cyclodextrin and pullulan.
机译：芽孢杆菌RM16是从卡拉奇温泉中分离出来的，并经过筛选用于生产α-淀粉酶。在含有1％淀粉（w / v）的Luria肉汤中培养菌株72小时后，获得了该酶。通过一系列连续步骤将酶Amy RM16纯化至电泳均一，包括用70％饱和硫酸铵沉淀，Q-Sepharose，Phenyl Sepharose和反相色谱分离。纯化的酶由一条66 kDa的多肽链组成，该多肽链是通过SDS-PAGE和酶谱分析相结合而建立的。在我们的实验条件下，获得的总收率为1.35％，比活性为6380U / mg，提供了17倍的最终酶纯化率。还进行了Amy RM16的生化表征，例如最佳温度和pH，底物特异性以及对不同金属离子和抑制剂的酶敏感性。这些研究结果表明，该酶在很宽的温度范围内都具有活性，在80°C时具有最佳活性，在50°C下3h保留85％的活性，而在80°C下1h保留约50％的活性。 。该酶在pH 5.0下显示最佳活性。另一方面，Ca 2+和EDTA（1-5mM）对酶活性没有显着影响。发现该酶的主要底物是淀粉，但它也可以水解未加工的淀粉，糊精，αg-环糊精和支链淀粉。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号