Production and characterization of recombinant lignin peroxidase isozyme H2 from Phanerochaete chrysosporium using recombinant baculovirus.
Production and characterization of recombinant lignin peroxidase isozyme H2 from Phanerochaete chrysosporium using recombinant baculovirus.
复制标题
使用重组杆状病毒从黄孢原毛平革菌中生产重组木质素过氧化物酶同工酶 H2 并进行表征。
DOI：
10.1016/0003-9861(92)90624-6
复制
发表时间：
1992
影响因子：
3.9
通讯作者：
Tien,M
中科院分区：
文献类型：
--
作者：
Johnson,TM;Pease,EA;Li,JK;Tien,M
用含有λML-6的基因工程杆状病毒感染昆虫细胞，产生黄孢原毛平革菌木质素过氧化物酶同工酶H2（pI4.4）。重组酶被纯化到接近同质，是能够氧化藜芦醇，碘化物，并在较小程度上，愈创木酚。重组酶对藜芦醇和H_2O_2的Km值与真菌酶相似。愈创木酚氧化活性或任何其它活性不依赖于Mn 2+。纯化的重组过氧化物酶用N-连接的碳水化合物糖基化。重组木质素过氧化物酶从阴离子交换树脂洗脱类似于天然同工酶H1，而不是H2。然而，重组酶的pI与H1和H2同工酶都不同。进一步表征天然同工酶H1和H2的真菌培养物显示相同的N-末端残基。这表明同工酶H1和H2在翻译后修饰方面不同。
RecombinantPhanerochaete chrysosporiumlignin peroxidase isozyme H2 (pI4.4) was produced in insect cells infected with a genetically engineered baculovirus containing a copy of the cDNA clone λML-6. The recombinant enzyme was purified to near homogeneity and is capable of oxidizing veratryl alcohol, iodide, and, to a lesser extent, guaiacol. TheKmof the recombinant enzyme for veratryl alcohol and H2O2is similar to that of the fungal enzyme. The guaiacol oxidation activity or any other activity is not dependent upon Mn2+. The purified recombinant peroxidase is glycosylated with N-linked carbohydrate(s). The recombinant lignin peroxidase eluted from an anion exchange resin similar to that of native isozyme H1 rather than H2. However, the pIof the recombinant enzymes is different from both H1 and H2 isozymes. Further characterization of native isozymes H1 and H2 from the fungal cultures revealed identical N-terminus residues. This indicates that isozymes H1 and H2 differ in post-translational modification.