从短小芽孢杆菌和类芽孢杆菌中纯化和部分表征锰过氧化物酶
Purification and Partial characterization of manganese peroxidase from Bacillus pumilus AND Paenibacillus sp.
机构信息
Divisão de Microbiologia, Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas, Universidade Estadual de Campinas , Campinas, SP , Brasil.
出版信息
Braz J Microbiol. 2009 Oct;40(4):818-26. doi: 10.1590/S1517-838220090004000012. Epub 2009 Dec 1.
The production of manganese peroxidase (MnP) from Bacillus pumilus and Paenibacillus sp. was studied under absence and presence of the inducers indulin AT, guayacol, veratryl alcohol, lignosulfonic acid and lignosulfonic acid desulfonated. Indulin AT increased the activity of B. pumilus MnP up to 31.66 U/L after 8 h, but no improve was observed for Paenibacillus sp., which reached maximum activity (12.22 U/L) after 20 h. Both MnPs produced by these microorganisms were purified in phenyl sepharose resin and the proteins from crude extracts were eluted in two fractions. However, only the first fraction of each extract exhibited MnP activities. Tests in different pH and temperature values, from pH 5.0 to pH 10.0 and 30 °C to 60 °C, respectively, were carried out with the purified MnP. The maximum activity reached for B. pumilus and Paenibacillus sp. MnPs were 4.3 U/L at pH 8.0 and 25 °C and 11.74 U/L at pH 9.0 and 35 °C, respectively. The molar masses determined by SDS-PAGE gel eletrophoresis were 25 kDa and 40 kDa, respectively, for the purified enzyme from B. pumilus and Paenibacillus sp.
研究了在不存在和存在诱导剂吲哚醇 AT、愈创木酚、藜芦醇、木质素磺酸钠和木质素磺酸钠脱磺基的情况下，从短小芽孢杆菌和巨大芽孢杆菌中生产锰过氧化物酶（MnP）的情况。吲哚醇 AT 在 8 小时后将短小芽孢杆菌 MnP 的活性提高到 31.66 U/L，但对巨大芽孢杆菌没有改善，后者在 20 小时后达到最大活性（12.22 U/L）。这两种微生物产生的 MnP 都在苯基琼脂糖树脂中进行了纯化，粗提物中的蛋白质在两个馏分中洗脱。然而，每个提取物的第一馏分仅表现出 MnP 活性。在不同的 pH 值和温度值（分别为 pH 5.0 至 pH 10.0 和 30°C 至 60°C）下对纯化的 MnP 进行了测试。短小芽孢杆菌和巨大芽孢杆菌 MnP 的最大活性分别为 pH 8.0 和 25°C 时的 4.3 U/L 和 pH 9.0 和 35°C 时的 11.74 U/L。通过 SDS-PAGE 凝胶电泳确定的分子量分别为 25 kDa 和 40 kDa，分别为从小短芽孢杆菌和巨大芽孢杆菌中纯化的酶。
Zotero 插件