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| 树状多节孢酸性磷酸酶的分离纯化与性质研究 | |
| 引用本文： | 张国庆,王有志,王守现,王贺祥,陈青君.树状多节孢酸性磷酸酶的分离纯化与性质研究[J].菌物学报,2011,30(5):753-759. |
| 作者姓名： | 张国庆 王有志 王守现 王贺祥 陈青君 |
| 作者单位： | 1. 北京农学院农业部都市农业(北方)重点实验室 北京102206;中国农业大学生物学院农业生物技术国家重点实验室 北京100193 2. 中国科学院微生物研究所菌种保藏中心 北京100101 3. 北京市农林科学院植物保护环境保护研究所 北京100097 4. 中国农业大学生物学院农业生物技术国家重点实验室 北京100193 5. 北京农学院农业部都市农业(北方)重点实验室 北京102206 |
| 基金项目： | 北京市科委项目（No. Z101105002510003）；北京市属高等学校人才强教项目（No. PHR200907136） |
| 摘 要： | 树状多节孢Nodulisporium sylviforme是从东北红豆杉Taxus cuspidata分离、可产生紫杉醇的内生真菌。研究以树状多节孢为材料，利用液体发酵手段获得菌丝体，通过CM-cellulose阴离子交换柱层析、Q-Sepharose阳离子交换柱层析和FPLC凝胶过滤层析（Superdex 75），获得纯化的树状多节孢酸性磷酸酶蛋白（Nod-ACP）。结合FPLC和SDS-PAGE分析，判定该磷酸酶为分子量44kDa单亚基蛋白。酶学性质研究表明，其最适pH值为3.0，最适温度为58℃。6 |
| 关 键 词： | 树状多节孢 酸性磷酸酶 性质 酶活性 |
Purification and characterization of an acid phosphatase from Nodulisporium sylviforme |
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| Authors: | ZHANG Guo-Qing WANG You-Zhi WANG Shou-Xian WANG He-Xiang and CHEN Qing-Jun |
| Institution: | Key Laboratory of Urban Agriculture (North) of Ministry of Agriculture, Beijing University of Agriculture, Beijing 102206, China State Key Laboratory for Agrobiotechnology, College of Biosciences, China Agricultural University, Beijing 100193, China;The Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;State Key Laboratory for Agrobiotechnology, College of Biosciences, China Agricultural University, Beijing 100193, China;Key Laboratory of Urban Agriculture (North) of Ministry of Agriculture, Beijing University of Agriculture, Beijing 102206, China |
| Abstract: | A taxol-producing endophytic fungus Nodulisporium sylviforme was isolated from Taxus cuspidata. In this study, a novel acid phosphatase was purified from the fungus by using a schedule of fermentation, ion exchange chromatography on CM-cellulose, Q-Sepharose and gel filtration by fast protein liquid chromatography on Superdex 75. The phosphatase was a monomeric protein with a molecular mass of 44kDa. Its optimum pH was 3.0 and optimum temperature 58℃. The activity of the phosphatase was enhanced by 30%－50% in the presence of Ca2+, Mn2+, Mg2+ and Zn2+ cations with the concentrations of 6.25－50mmol/L, and was reduced in the presence of Cu2+, Al3+ and Fe3+ cations. Fe2+ increased the enzyme activity at the concentration lower than 25mmol/L, but inhibited the enzyme activity completely at the concentration more than 50mmol/L. |
| Keywords: | Nodulisporium sylviforme acid phosphatase characteristics enzyme activity |
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