Purification and Partial characterization of manganese peroxidase from Bacillus pumilus AND Paenibacillus sp.
Purification and Partial characterization of manganese peroxidase from Bacillus pumilus AND Paenibacillus sp.
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DOI：
10.1590/s1517-83822009000400012
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发表时间：
2009-12-01
影响因子：
2.2
通讯作者：
Durrant, Lúcia Regina
中科院分区：
文献类型：
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作者：
Oliveira, Patrícia Lopes de;Duarte, Marta Cristina Teixeira;Durrant, Lúcia Regina
从短小芽孢杆菌和类芽孢杆菌属中生产锰过氧化物酶（MnP）。在诱导剂胰岛素AT、愈创木酚、藜芦醇、木质素磺酸和脱磺化木质素磺酸不存在和存在的情况下进行了研究。 8 小时后，Indulin AT 将短小芽孢杆菌 MnP 的活性提高至 31.66 U/L，但类芽孢杆菌没有观察到任何改善，类芽孢杆菌在 20 小时后达到最大活性（12.22 U/L）。这些微生物产生的两种 MnP 均在苯基琼脂糖树脂中纯化，粗提物中的蛋白质以两部分洗脱。然而，只有每种提取物的第一部分表现出 MnP 活性。使用纯化的MnP分别在不同pH值和温度值（pH 5.0至pH 10.0和30℃至60℃）下进行测试。 B. pumilus 和 Paenibacillus sp. 达到最大活性。在 pH 8.0 和 25 摄氏度下，MnP 分别为 4.3 U/L；在 pH 9.0 和 35 摄氏度下，MnP 分别为 11.74 U/L。对于来自短小芽孢杆菌和类芽孢杆菌属的纯化酶，通过 SDS-PAGE 凝胶电泳测定的摩尔质量分别为 25 kDa 和 40 kDa。
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 degrees C to 60 degrees 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 degrees C and 11.74 U/L at pH 9.0 and 35 degrees 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.