Molecular characterization of aromatic peroxygenase from Agrocybe aegerita
Molecular characterization of aromatic peroxygenase from Agrocybe aegerita
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DOI：
10.1007/s00253-009-2000-1
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发表时间：
2009-10-01
影响因子：
5
通讯作者：
Hofrichter, Martin
中科院分区：
文献类型：
--
作者：
Pecyna, Marek J.;Ullrich, Rene;Hofrichter, Martin
最近发现了一组新的真菌过氧化物酶，称为芳香族过氧化物酶(APO)。这些由蘑菇担子菌产生的这些胞外生物催化剂的成员可以催化反应--例如，萘、甲苯、二苯并噻吩苯或吡啶的过氧化反应--这些反应实际上归因于细胞色素P450单加氧酶。在这里，第一次提出了关于这组新的过氧化氢消耗酶的遗传信息。从信使RNA和基因组DNA水平对山羊草过氧化物酶基因(Apo1)进行了鉴定。纯化酶的Edman降解和从头测序得到的多肽序列证实了该基因的序列。实时定量逆转录聚合酶链式反应表明，山羊草apo1酶活性的变化过程与其mRNA信号的变化过程具有很好的相关性。山羊草过氧酶的全长序列和弧度弯曲霉过氧酶的部分序列证实了这些酶与血红素硫酸盐蛋白的从属关系。这些序列与经典的过氧化物酶、细胞色素P450酶没有同源性，与子囊菌Caldarimyces fumago产生的真菌氯过氧化物酶只有很少的同源性(<；30%)(这只在蛋白质的N端部分，包括血红素结合区和部分远端的血红素口袋)。这一事实加强了APO蛋白的新颖性。另一方面，在遗传数据库中的同源检索导致了各种APO同源基因和转录本的鉴定，特别是在黑木耳真菌中，表明APO在真菌王国中广泛存在。
Recently, a novel group of fungal peroxidases, known as the aromatic peroxygenases (APO), has been discovered. Members of these extracellular biocatalysts produced by agaric basidiomycetes such as Agrocybe aegerita or Coprinellus radians catalyze reactions-for example, the peroxygenation of naphthalene, toluene, dibenzothiophene, or pyridine-which are actually attributed to cytochrome P450 monooxygenases. Here, for the first time, genetic information is presented on this new group of peroxide-consuming enzymes. The gene of A. aegerita peroxygenase (apo1) was identified on the level of messenger RNA and genomic DNA. The gene sequence was affirmed by peptide sequences obtained through an Edman degradation and de novo peptide sequencing of the purified enzyme. Quantitative real-time reverse transcriptase polymerase chain reaction demonstrated that the course of enzyme activity correlated well with that of mRNA signals for apo1 in A. aegerita. The full-length sequences of A. aegerita peroxygenase as well as a partial sequence of C. radians peroxygenase confirmed the enzymes' affiliation to the heme-thiolate proteins. The sequences revealed no homology to classic peroxidases, cytochrome P450 enzymes, and only little homology (< 30%) to fungal chloroperoxidase produced by the ascomycete Caldariomyces fumago (and this only in the N-terminal part of the protein comprising the heme-binding region and part of the distal heme pocket). This fact reinforces the novelty of APO proteins. On the other hand, homology retrievals in genetic databases resulted in the identification of various APO homologous genes and transcripts, particularly among the agaric fungi, indicating APO's widespread occurrence in the fungal kingdom.