High-yield production of aromatic peroxygenase by the agaric fungus Marasmius rotula.
High-yield production of aromatic peroxygenase by the agaric fungus Marasmius rotula.
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DOI：
10.1186/2191-0855-1-31
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发表时间：
2011-10-11
期刊：
影响因子：
3.7
通讯作者：
Scheibner K
中科院分区：
文献类型：
--
作者：
Gröbe G;Ullrich R;Pecyna MJ;Kapturska D;Friedrich S;Hofrichter M;Scheibner K
采用液体培养法从圆小皮伞（Marasmius rotula）中产生一种胞外过氧合酶，经层析纯化并进行部分表征。这是第三个芳香族过氧化酶（APO），已详细的特点和第一个可以高产率生产。最高酶水平约为41，000 U l-1（对应于约100 U l-1）。445毫克l-1载脂蛋白）超过迄今报道的水平超过40倍，并在富碳和氮的复杂介质中检测到。通过FPLC将酶纯化至表观均一性（SDS-PAGE），分子量为32 kDa（去糖基化后为27 kDa），等电点在4.97和5.27之间。天然酶的UV-可见光谱在418 nm处显示出特征最大值（Soret带），其在用连二亚硫酸钠还原并用一氧化碳冲洗后移位至443 nm。最适pH值为2.0。对于所研究的大多数反应，发现圆酵母酶在pH 5和6之间变化。2，6-二甲氧基苯酚、苯甲醇、藜芦醇、萘和H2 O2的表观Km值分别为0.133、0.118、0.279、0.791和3.14 mM。M.发现圆叶APO在5 - 10的pH范围内以及在有机溶剂（50%vol/vol）如甲醇、乙腈和N，N-二甲基甲酰胺存在下高度稳定。与其它APO不同，M. rotula既不表现出溴化活性也不表现出氯化活性。
An extracellular peroxygenase from Marasmius rotula was produced in liquid culture, chromatographically purified and partially characterized. This is the third aromatic peroxygenase (APO) that has been characterized in detail and the first one that can be produced in high yields. The highest enzyme levels of about 41,000 U l-1 (corresponding to appr. 445 mg l-1 APO protein) exceeded the hitherto reported levels more than 40-fold and were detected in carbon- and nitrogen-rich complex media. The enzyme was purified by FPLC to apparent homogeneity (SDS-PAGE) with a molecular mass of 32 kDa (27 kDa after deglycosylation) and isoelectric points between 4.97 and 5.27. The UV-visible spectrum of the native enzyme showed a characteristic maximum (Soret band) at 418 nm that shifted after reduction with sodium dithionite and flushing with carbon monoxide to 443 nm. The pH optimum of the M. rotula enzyme was found to vary between pH 5 and 6 for most reactions studied. The apparent Km-values for 2,6-dimethoxyphenol, benzyl alcohol, veratryl alcohol, naphthalene and H2O2 were 0.133, 0.118, 0.279, 0.791 and 3.14 mM, respectively. M. rotula APO was found to be highly stable in a pH range from 5 to 10 as well as in the presence of organic solvents (50% vol/vol) such as methanol, acetonitrile and N,N-dimethylformamide. Unlike other APOs, the peroxygenase of M. rotula showed neither brominating nor chlorinating activities.
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影响因子：
5
作者：
Pecyna, Marek J.;Ullrich, Rene;Hofrichter, Martin
通讯作者：
Hofrichter, Martin
影响因子：
2.8
作者：
Celio, G. J.;Padamsee, M.;McLaughlin, D. J.
通讯作者：
McLaughlin, D. J.
影响因子：
4.4
作者：
Anh, Dau Hung;Ullrich, Rene;Hofrichter, Martin
通讯作者：
Hofrichter, Martin
影响因子：
5
作者：
Majcherczyk, A;Johannes, C;Hüttermann, A
通讯作者：
Hüttermann, A
DOI：
10.1073/pnas.91.18.8710
发表时间：
1994-08-30
影响因子：
11.1
作者：
SMITH, GCM;TEW, DG;WOLF, CR
通讯作者：
WOLF, CR