Departamento de Alimentos e Nutrição, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista , Araraquara, SP , Brasil.
Braz J Microbiol. 2008 Oct;39(4):682-8. doi: 10.1590/S1517-838220080004000016. Epub 2008 Dec 1.
Three strains of Bacillus sp. (BACRP, BACNC-1 and BACAR) were isolated from soil adhered to cassava husk. CGTase specific activity for the three isolated strains was higher when cultivated at 40°C. Potato starch, cassava starch, maltodextrin and glucose were used as carbon source and growth temperatures varied from 25 to 55°C. The three isolates presented higher CGTase specific activity when cultivated with potato starch at 40°C. Isolated BACRP and BACAR presented specific activity of 4.0×10(-3) and 2.2×10(-3) U/mg prot at pH 7.0, respectively, when cultivated in mediums added with NaCl 2%; at pH 10,0 their activities were of 3.4×10(-3) and 3.0×10(-3) U/mg prot, respectively, in the same concentration of NaCl. On the other hand, the isolated BACNC-1 presented activity specific of 2.4×10(-3) U/mg prot when cultivated at pH 7.0 added of NaCl 1%, and at pH 10.0 the specific activity was of 3.4×10(-3) U/mg prot without NaCl addition. This work also showed the presence of cyclodextrins formed during fermentation process and that precipitation with acetone or lyophilization followed by dialysis was efficient at removing CDs (cyclodextrins), thus, eliminating interference in the activity assays. The enzyme produced by the BACAR strain was partially purified and β-CD was liberated as a reaction product.
从木薯皮上的土壤中分离到三株芽孢杆菌(BACRP、BACNC-1 和 BACAR)。这三株分离菌在 40°C 下培养时,CGTase 比活更高。以马铃薯淀粉、木薯淀粉、麦芽糊精和葡萄糖为碳源,生长温度从 25°C 到 55°C 不等。这三株分离菌在 40°C 下以马铃薯淀粉为碳源时,CGTase 比活更高。当在添加 2%NaCl 的培养基中培养时,分离得到的 BACRP 和 BACAR 的比活分别为 4.0×10(-3)和 2.2×10(-3)U/mg 蛋白,在 pH 10.0 时,其活性分别为 3.4×10(-3)和 3.0×10(-3)U/mg 蛋白,在相同浓度的 NaCl 下。另一方面,分离得到的 BACNC-1 在添加 1%NaCl 的 pH 7.0 培养基中培养时,比活为 2.4×10(-3)U/mg 蛋白,在 pH 10.0 时,无 NaCl 存在时,比活为 3.4×10(-3)U/mg 蛋白。这项工作还表明,在发酵过程中存在形成的环糊精,用丙酮沉淀或冻干后透析可以有效地去除 CDs(环糊精),从而消除了对活性测定的干扰。从 BACAR 菌株中生产的酶进行了部分纯化,β-CD 作为反应产物被释放出来。