Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii

›› 2010, Vol. 18 ›› Issue (5): 824-829.

Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii

陈敏^1,2, 姚善泾¹, 张虹¹, 梁新乐²

1. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China;
2. Department of Biotechnology Engineering, Zhejiang Gongshang University, Hangzhou 310012, China

收稿日期:2009-12-29 修回日期:2010-05-01 出版日期:2010-10-28 发布日期:2010-10-28
通讯作者: YAO Shanjing,E-mail:yaosj@zju.edu.cn
基金资助:
Supported by the Special Funds for Major State Basic Research Program of China (2007CB707805);the Natural Science Foundation of Zhejiang Province (Y505334)

Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii

CHEN Min^1,2, YAO Shanjing¹, ZHANG Hong¹, LIANG Xinle²

1. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China;
2. Department of Biotechnology Engineering, Zhejiang Gongshang University, Hangzhou 310012, China

Received:2009-12-29 Revised:2010-05-01 Online:2010-10-28 Published:2010-10-28
Supported by:
Supported by the Special Funds for Major State Basic Research Program of China (2007CB707805);the Natural Science Foundation of Zhejiang Province (Y505334)

摘要/Abstract

摘要： A versatile peroxidase (VP_-Peco60-7) was generated and purified from the liquid culture of Pleurotus eryngii. The purification procedure included ammonium sulfate precipitation, ion exchange chromatography, and gel chromatography. The molecular weight and isoelectric point (pI) of VP_-Peco60-7 were determined to be approxi-mately 40 kDa and 4.1, respectively. By N-terminal sequence determination and peptide mapping analysis, VP_-Peco60-7 was found to be similar to the versatile peroxidase isoenzyme VPL1, which was previously isolated from liquid cultures of the same species. However, the molecular weight and pI of VP_-Peco60-7 were different from those of versatile peroxidases of liquid cultures, implying that the VP_-Peco60-7 in this study is of a novel type. With 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) as a substrate, the maximal enzyme activity was obtained at 50℃ and pH 3.0. The catalysis of ABTS by VP_-Peco60-7 was expressed by the Michaelis-Menten equa-tion. At 50℃ and pH 3.0, the maximum velocity (V_max) was 188.68 U·mg^-1 and the michaelis constant (K_m) was 203.09 μmol·L^-1.

关键词: Pleurotus eryngii, versatile peroxidase, purification, enzymatic properties

Abstract: A versatile peroxidase (VP_-Peco60-7) was generated and purified from the liquid culture of Pleurotus eryngii. The purification procedure included ammonium sulfate precipitation, ion exchange chromatography, and gel chromatography. The molecular weight and isoelectric point (pI) of VP_-Peco60-7 were determined to be approxi-mately 40 kDa and 4.1, respectively. By N-terminal sequence determination and peptide mapping analysis, VP_-Peco60-7 was found to be similar to the versatile peroxidase isoenzyme VPL1, which was previously isolated from liquid cultures of the same species. However, the molecular weight and pI of VP_-Peco60-7 were different from those of versatile peroxidases of liquid cultures, implying that the VP_-Peco60-7 in this study is of a novel type. With 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) as a substrate, the maximal enzyme activity was obtained at 50℃ and pH 3.0. The catalysis of ABTS by VP_-Peco60-7 was expressed by the Michaelis-Menten equa-tion. At 50℃ and pH 3.0, the maximum velocity (V_max) was 188.68 U·mg^-1 and the michaelis constant (K_m) was 203.09 μmol·L^-1.

Key words: Pleurotus eryngii, versatile peroxidase, purification, enzymatic properties

陈敏, 姚善泾, 张虹, 梁新乐. Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii[J]. , 2010, 18(5): 824-829.

CHEN Min, YAO Shanjing, ZHANG Hong, LIANG Xinle. Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii[J]. , 2010, 18(5): 824-829.

参考文献

1 Ward, G., Hadar, Y., Dosoretz, C.G., “The biodegradation of lignocellulose by white rot fungi”, In:Fungal Biotechnology in Agricultural, Food, and Environmental Applications, Dilip K. Arora, ed., Marcel Dekker Press, New York, 393-407 (2004).
2 Cripps, C., Bumpus, J.A., Aust, S.D., “Biodegradation of azo and heterocyclic dyes by Phanerochaete chrysosporium”, Appl. Environ. Microbiol., 56, 1114-1118 (1990).
3 Sutherland, J.B., “Degradation of hydrocarbons by yeast and filamentous fungi”, In:Degradation of Hydrocarbons by Yeast and Filamentous Fungi, Marcel Dekker Press, New York, 443-455 (2004).
4 Faull, J.L., Baker, S.C., Wilkinson, S., Nicklin, S., In:Fungal Degradation of Explosives, Marcel Dekker Press, New York, 471-480 (2004).
5 Li, H.R., Biology and Biotechnology of White Rot Fungi, Chemical Industry Press , Beijing, 25-58 (2005).
6 Martínez, M.J., Ruiz-Due as, F.J., Guillén, F., Martínez, A.T., “Purification and catalytic properties of two manganese peroxidase isoenzymes from Pleurotus eryngii”, Eur. J. Biochem., 237, 424-432 (1996).
7 Heinfling, A., Ruiz-Due as, F.J., Martínez, M.J., Bergbauer, M., Szewzyk, U., Martínez, A.T., “A study on reducing substrates of manganese-oxidizing peroxidases from Pleurotus eryngii and Bjerkandera adusta”, FEBS. Lett., 428, 141-146 (1998).
8 Moreira, P.R., Duez, C., Dehareng, D., Antunes, A., Almeida-Vara, E., Frere, J.M., Malcata, F.X., Duarte, J.C., “Molecular characterization of a versatile peroxidase from a Bjekandera strain”, J. Biotechnol., 118, 339-352 (2005).
9 Pogni, R., Baratto, M.C., Giansanti, S., Teutloff, C., Verdin, J., Valderrama, B., Lendzian, F., Lubitz, W., Vazquez-Duhalt, R., Basosi, R., “Tryptophan-based radical in the catalytic mechanism of versatile peroxidase from Bjerkandera adusta”, Biochemistry, 44, 4267-4274 (2005).
10 Martínez, A.T., Camarero, S., Guillén, F., Gutiérrez, A., Mu oz, C., Varela, E., Martínez, M.J., Barrasa, J.M., Ruel, K., Pelayo, M., “Progress in biopulping of non-woody materials:Chemical, enzymatic and ultrastructural aspects of wheat straw delignification with ligninolytic fungi from the genus Pleurotus”, FEMS. Microbiol. Rev., 13, 265-274 (1994).
11 Camarero, S., Barrasa, J.M., Pelayo, M., Martínez, A.T., “Evaluation of pleurotus species for wheat straw biopulping”, J. Pulp. Paper. Sci., 24, 197-203 (1998).
12 Camarero, S., Sarkar, S., Ruiz-Due as, F.J., Martínez, M.J., Martinez, A.T., “Description of a versatile peroxidase involved in the natural degradation of lignin that has both manganese peroxidase and lignin peroxidase substrate interaction sites”, J. Biol. Chem., 274, 10324-10330 (1999).
13 Caramelo, L., Martínez, M.J., Martnez, A.T., “A search for ligninolytic peroxidases in the fungus Pleurotus eryngii involving α-keto-γ-thiomethylbutyric acid and lignin model dimmers”, Appl. Environ. Microbiol., 65, 916-922 (1999).
14 Ruiz-Due as, F.J., Martínez, M.J., Martínez, A.T., “Molecular characterization of a novel peroxidase isolated from ligninolytic fungus Pleurotus eryngii”, Mol. Microbiol., 31, 223-235 (1999).
15 Camarero, S., Ruiz-Due as, F.J., Sarkar, S., Martínez, M.J., Martínez, A.T., “The cloning of a new peroxidase found in lignocellulose cultures of Pleurotus eryngii and sequence comparison with other fungal peroxidases”, FEMS. Microbiol. Lett., 191, 37-43 (2000).
16 Ruiz-Due as, F.J., Martínez, M.J., Martínez, A.T., “Heterologous expression of Pleurotus eryngii peroxidase confirms its ability to oxidize Mn²⁺ and different aromatic substrates”, Appl. Environ. Microbiol., 65, 4705-4707 (1999).
17 Lú-Chau, T.A., Ruiz-Due as, F.J., Camarero, S., Feijoo, G., Martínez, M.J., Lema, J.M., Martínez, A.T., “Effect of pH on the stability of Pleurotus eryngii versatile peroxidase during heterologous production in Emericella nidulan”, Bioprocess. Biosyst. Eng., 26, 287-293 (2004).
18 Eibes, G.M., Lú-Chau, T.A., Ruiz-Due as, F.J., Feijoo, G., Martínez, M.J., Martínez, A.T., Lema, J.M., “Effect of culture temperature on the heterologous expression of Pleurotus eryngii versatile peroxidase in Aspergillus hosts”, Bioprocess. Biosyst. Eng., 32, 129-134 (2009).
19 Munoz, C., Guillen, F., Martinez, A.T., Martinez, M.J., “Laccase isoenzymes of Pleurotus eryngii:characterization, catalytic properties, and participation in activation of molecular oxygen and Mn²⁺ oxidation”, Appl. Envir. Microbiol., 63, 21662174 (1997).
20 Laemmli, U.K., “Cleavage of structural proteins during the assembly of the head of bacteriophage T4”, Nature, 227, 680-685 (1970).
21 Yu, R.Y., Yuan, M.X., Chen, L.R., “Principle and methods of biochemistry experiment”, In:Principle and Methods of Biochemistry Experiment, Peking University Press, Beijing, 240-323 (2008).
22 Li, L., Zhang, X.C., Xu, M.H., “Study on dynamics of Mn peroxidase (MnP) by white rot fungi”, Journal of Anhui University of Science and Technology (Natural Science), 25, 63-67 (2005).
23 Camarero, S., B ckle, B., Martínez, M.J., Martínez, A.T., “Manganese-mediated lignin degradation by Pleurotus pulmonarius”, Appl. Environ. Microbiol., 62, 1070-1072 (1996).
24 Sarkar, S., Martínez, A.T., Martínez, M.J., “Biochemical and molecular characterization of manganese peroxidase isoenzyme from Pleurotus ostreatus”, Biochem. Biophys. Acta., 1339, 23-30 (1997).
25 Gómez-Toribio, V., Martínez, A.T., Martínez, M.J., Guillén, F., “Oxidation of hydroquinones by the versatile ligninolytic peroxidase from Pleurotus eryngii-H₂O2 generation and the influence of Mn²⁺”, Eur. J. Biochem., 268, 4787-4793 (2001).

[1]	Yuan Liu, Hanting Xiong, Jingwen Chen, Shixia Chen, Zhenyu Zhou, Zheling Zeng, Shuguang Deng, Jun Wang. One-step ethylene separation from ternary C₂ hydrocarbon mixture with a robust zirconium metal-organic framework[J]. 中国化学工程学报, 2023, 59(7): 9-15.
[2]	Yong Xu, Qingbai Chen, Yang Gao, Jianyou Wang, Huiqing Fan, Fei Zhao. Performance comparison of lithium fractionation from magnesium via continuous selective nanofiltration/electrodialysis[J]. 中国化学工程学报, 2023, 59(7): 42-50.
[3]	Runze Chen, Yuran Chen, Xuemin Liang, Yapeng Kong, Yangyang Fan, Quan Liu, Zhenyu Yang, Feiying Tang, Johnny Muya Chabu, Maru Dessie Walle, Liqiang Wang. Oxidative exfoliation of spent cathode carbon: A two-in-one strategy for its decontamination and high-valued application[J]. 中国化学工程学报, 2023, 59(7): 262-269.
[4]	Yunchang Fan, Chunyan Zhu, Sheli Zhang, Lei Zhang, Qiang Wang, Feng Wang. Efficient and selective extraction of sinomenine by deep eutectic solvents[J]. 中国化学工程学报, 2023, 57(5): 109-117.
[5]	Juanjuan Liu, Xiaolong Lu, Guiming Shu, Ke Li, Shuyun Zheng, Xiao Kong, Tao Li, Jun Yang. The facile method developed for preparing polyvinylidene fluoride plasma separation membrane via macromolecular interaction[J]. 中国化学工程学报, 2022, 49(9): 140-149.
[6]	Wenjian Zhu, Xuhua Shen, Rui Ou, Manoj Murugesan, Aihua Yuan, Jianfeng Liu, Xiaocai Hu, Zhen Yang, Ming Shen, Fu Yang. Superhigh selective capture of volatile organic compounds exploiting cigarette butts-derived engineering carbonaceous adsorbent[J]. 中国化学工程学报, 2022, 46(6): 194-206.
[7]	Song Hu, Jinlong Li, Qihua Wang, Weisheng Yang. Design and optimization of an integrated process for the purification of propylene oxide and the separation of propylene glycol by-product[J]. 中国化学工程学报, 2022, 45(5): 111-120.
[8]	Puxu Liu, Yong Wang, Yang Chen, Xiaoqing Wang, Jiangfeng Yang, Libo Li, Jinping Li. Stable titanium metal-organic framework with strong binding affinity for ethane removal[J]. 中国化学工程学报, 2022, 42(2): 35-41.
[9]	Xionghui Liu, Jianfeng Du, Yu Ye, Yuchuan Liu, Shun Wang, Xianyu Meng, Xiaowei Song, Zhiqiang Liang, Wenfu Yan. Boosting selective C₂H₂/CH₄, C₂H₄/CH₄ and CO₂/CH₄ adsorption performance via 1,2,3-triazole functionalized triazine-based porous organic polymers[J]. 中国化学工程学报, 2022, 42(2): 64-72.
[10]	Tiefeng Wang, Jinxiang Dong. Ethylene glycol purification by melt crystallization: Removal of 2-methoxyethanol impurity[J]. 中国化学工程学报, 2021, 37(9): 39-45.
[11]	Piqiang Tan, Xiaoyu Li, Shiyan Wang, Zhiyuan Hu, Diming Lou. Selective catalytic reduction failure of low NH₃-NO_x ratio[J]. 中国化学工程学报, 2021, 32(4): 231-240.
[12]	Kai Zhu, Chaoqun Yao, Yanyan Liu, Guangwen Chen. Using expansion units to improve CO₂ absorption for natural gas purification-A study on the hydrodynamics and mass transfer[J]. 中国化学工程学报, 2021, 29(1): 35-46.
[13]	Xue Gao, Sisi Wen, Bingling Yang, Jian Xue, Haihui Wang. Enhanced air filtration performance under high-humidity condition through electrospun membranes with optimized structure[J]. 中国化学工程学报, 2020, 28(7): 1788-1795.
[14]	Xinping Li, Yaowei Wang, Tangyin Wu, Shichao Song, Bin Wang, Shenglai Zhong, Rongfei Zhou. High-performance SSZ-13 membranes prepared using ball-milled nanosized seeds for carbon dioxide and nitrogen separations from methane[J]. 中国化学工程学报, 2020, 28(5): 1285-1292.
[15]	Jing Guo, Weizhou Jiao, Guisheng Qi, Zhiguo Yuan, Youzhi Liu. Applications of high-gravity technologies in gas purifications: A review[J]. 中国化学工程学报, 2019, 27(6): 1361-1373.

Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii

Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价