Genome-to-function characterization of novel fungal P450 monooxygenases oxidizing polycyclic aromatic hydrocarbons (PAHs).
Genome-to-function characterization of novel fungal P450 monooxygenases oxidizing polycyclic aromatic hydrocarbons (PAHs).
复制标题
DOI：
10.1016/j.bbrc.2010.07.094
复制
发表时间：
2010-09-03
影响因子：
3.1
通讯作者：
Yadav, Jagjit S.
中科院分区：
文献类型：
--
作者：
Syed, Khajamohiddin;Doddapaneni, Harshavardhan;Subramanian, Venkataramanan;Lam, Ying Wai;Yadav, Jagjit S.
关键词：
Fungi, particularly the white rot basidiomycetes, have an extraordinary capability to degrade and/or mineralize (to CO2) the recalcitrant fused-ring high molecular weight (≥ 4 aromatic-rings) polycyclic aromatic hydrocarbons (HMW PAHs). Despite over 30 years of research demonstrating involvement of P450 monooxygenation reactions in fungal metabolism of HMW PAHs, specific P450 monooxygenases responsible for oxidation of these compounds are not yet known. Here we report the first comprehensive identification and functional characterization of P450 monooxygenases capable of oxidizing different ring-size PAHs in the model white rot fungus Phanerochaete chrysosporium using a successful genome-to-function strategy. In a genome-wide P450 microarray screen, we identified six PAH-responsive P450 genes (Pc-pah1 - Pc-pah6) inducible by PAHs of varying ring size, namely naphthalene, phenanthrene, pyrene, and benzo(a)pyrene (BaP). Using a co-expression strategy, cDNAs of the six Pc-Pah P450s were cloned and expressed in Pichia pastoris in conjunction with the homologous P450 oxidoreductase (Pc-POR). Each of the six recombinant P450 monooxygenases showed PAH-oxidizing activity albeit with varying substrate specificity towards PAHs (3–5 rings). All six P450s oxidized pyrene (4-ring) into two monohydroxylated products. Pc-Pah1 and Pc-Pah3 oxidized BaP (5-ring) to 3-hydroxyBaP whereas Pc-Pah4 and Pc-Pah6 oxidized phenanthrene (3-ring) to 3-, 4-, and 9-phenanthrol. These PAH-oxidizing P450s (493 – 547 aa) are structurally diverse and novel considering their low overall homology (12–23%) to mammalian counterparts. To our knowledge, this is the first report on specific fungal P450 monooxygenases with catalytic activity toward environmentally persistent and highly toxic HMW PAHs.
登录
查看更多内容
影响因子：
2.4
作者：
TATARKO, M;BUMPUS, JA
通讯作者：
BUMPUS, JA
影响因子：
4.4
作者：
Park J;Lee S;Choi J;Ahn K;Park B;Park J;Kang S;Lee YH
通讯作者：
Lee YH
影响因子：
4.1
作者：
Bernhardt, Rita
通讯作者：
Bernhardt, Rita
影响因子：
4.4
作者：
Bezalel, L;Hadar, Y;Cerniglia, CE
通讯作者：
Cerniglia, CE
DOI：
10.1016/j.bbrc.2009.06.134
发表时间：
2009-09-11
影响因子：
3.1
作者：
Kasai, Noriyuki;Ikushiro, Shin-ichi;Sakaki, Toshiyuki
通讯作者：
Sakaki, Toshiyuki