高等学校化学学报

高等学校化学学报

• 研究论文 • 上一篇    下一篇

原位光谱法检测白腐真菌P450的诱导及其降解功能

宁大亮, 王慧, 庄芫, 李冬   

  1. 清华大学环境科学与工程系, 环境模拟与污染控制国家重点联合实验室, 北京 100084
  • 收稿日期:2006-10-25 修回日期:1900-01-01 出版日期:2007-08-10 发布日期:2007-08-10
  • 通讯作者: 王慧

Inducement and Degradation Function of P450 in Phanerochaete chrysosporium by in Situ Spectroscopic Analysis

NING Da-Liang, WANG Hui*, ZHUANG Yuan, LI Dong   

  1. State Key Joint Laboratory on Environment Simulation and Pollution Control, Department of Environmental Science and Engineering, Tsinghua University, Beijing 100084, China
  • Received:2006-10-25 Revised:1900-01-01 Online:2007-08-10 Published:2007-08-10
  • Contact: WANG Hui

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被引次数

9

摘要： 利用诱导-原位光谱分析法, 对黄孢原毛平革菌中的微粒体P450在蛋白质水平上进行原位直接检测和表征, 研究结果表明, 正己烷、苯甲酸、萘和五氯酚对黄孢原毛平革菌P450酶系均有不同程度的诱导作用, 在合适的诱导剂量和诱导条件下, 对黄孢原毛平革菌P450的诱导量分别达到未诱导时的7.5, 5.6, 3.1和3.5 倍. 其中五氯酚对真菌P450酶系的诱导现象尚未见报道. 在不同的培养条件下测定了苯甲酸与P450的底物结合光谱, 并检测出微粒体酶液中依赖NADPH的苯甲酸降解活性, 证实了降解苯甲酸的P450基因在黄孢原毛平革菌中能够顺利合成并表现出明显的催化活性, 抑制剂实验结果表明, 除了P450的催化降解途径以外, 白腐真菌中还存在其它的苯甲酸降解途径.

关键词: CO结合差光谱, 白腐真菌, P450, 诱导, 降解功能

Abstract: The method of inducement-in situ spectra analysis was developed and used to directly detect and characterize microsome P450 and its inducement in Phanerochaete chrysosporium(PC) by n-hexane, benzoic acid, naphthalene and pentachlorophenol at molecular level. The results demonstrate that the content of P450 after inducement by n-hexane, benzoic acid, naphthalene reached a high level of 7.5, 5.6 and 3.1 times of that without inducement, respectively. The inducement of P450 in fungi by pentachlorophenol has not been reported before this study. The substrate binding spectra of benzoic acid with P450 from PC grown in both low nitrogen medium and PDB medium, and NADPH-dependent degradation of benzoic acid in PC microsome were detected, which provide a direct evidence of the function of P450 to catabolize benzoic acid. However, the inhibition was not observed during the degradation of benzoic acid by PC with P450 inhibitor aminobenzotriazole, which indicates that other catabolic pathway may also exist in PC.

Key words: Reduced carbon monooxide binding difference spectrum, Phanerochaete chrysosporium, P450, Inducement, Degradation

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