DyP-like peroxidases of the jelly fungus Auricularia auricula-judae oxidize nonphenolic lignin model compounds and high-redox potential dyes
DyP-like peroxidases of the jelly fungus Auricularia auricula-judae oxidize nonphenolic lignin model compounds and high-redox potential dyes
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DOI：
10.1007/s00253-009-2173-7
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发表时间：
2010-02-01
影响因子：
5
通讯作者：
Hofrichter, Martin
中科院分区：
文献类型：
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作者：
Liers, Christiane;Bobeth, Caroline;Hofrichter, Martin
果冻真菌黑木耳，judae产生的酶与锰独立的过氧化物酶活性在山毛榉木生长过程中（类似于300 U l（-1））。检测到相同的酶活性，并在包含液体、植物基培养基（例如番茄汁悬浮液）的搅拌培养物中以高达8，000 μ l（-1）的水平大规模产生。两种纯过氧化物酶形式（A.通过三个层析步骤可以从各自的培养液中获得黑木耳过氧化物酶（AjP I和AjP II）。光谱和电泳分析纯化的蛋白质揭示了它们的血红素和过氧化物酶的性质。AjP的N-末端氨基酸序列与被称为“染料脱色过氧化物酶”的真菌酶的序列匹配良好。与小皮伞（Marasmius scorodonius）、黄瓜疫霉（Thanatephorus cucumeris）和鸡枞菌（Termitomyces albuminosus）的过氧化物酶N-末端同源性分别为86%、60%和60%。两种酶形式不仅催化典型过氧化物酶底物如2，6-二甲氧基苯酚和2，2 '-连氮基-双（3-乙基噻唑啉-6-磺酸盐）的转化，而且催化高氧化还原电位染料活性蓝5和活性黑5的脱色，而锰（II）离子（Mn 2+）不被氧化。然而，最值得注意的是，这两个AjPs氧化非酚类木质素模型化合物（藜芦醇; adlerol，一种非酚类β-O-4木质素模型二聚体）在低pH（最大活性在pH 1.4），这表明一定的木质素分解活性的染料脱色过氧化物酶的发现。
The jelly fungus Auricularia auricula-judae produced an enzyme with manganese-independent peroxidase activity during growth on beech wood (similar to 300 U l(-1)). The same enzymatic activity was detected and produced at larger scale in agitated cultures comprising of liquid, plant-based media (e.g. tomato juice suspensions) at levels up to 8,000 U l(-1). Two pure peroxidase forms (A. auricula-judae peroxidase (AjP I and AjP II) could be obtained from respective culture liquids by three chromatographic steps. Spectroscopic and electrophoretic analyses of the purified proteins revealed their heme and peroxidase nature. The N-terminal amino acid sequence of AjP matched well with sequences of fungal enzymes known as "dye-decolorizing peroxidases". Homology was found to the N-termini of peroxidases from Marasmius scorodonius (up to 86%), Thanatephorus cucumeris (60%), and Termitomyces albuminosus (60%). Both enzyme forms catalyzed not only the conversion of typical peroxidase substrates such as 2,6-dimethoxyphenol and 2,2'-azino-bis(3-ethylthiazoline-6-sulfonate) but also the decolorization of the high-redox potential dyes Reactive Blue 5 and Reactive Black 5, whereas manganese(II) ions (Mn2+) were not oxidized. Most remarkable, however, is the finding that both AjPs oxidized nonphenolic lignin model compounds (veratryl alcohol; adlerol, a nonphenolic beta-O-4 lignin model dimer) at low pH (maximum activity at pH 1.4), which indicates a certain ligninolytic activity of dye-decolorizing peroxidases.