高等学校化学学报

高等学校化学学报 ›› 2001, Vol. 22 ›› Issue (7): 1137.

• 研究论文 • 上一篇    下一篇

光合细菌Chromatium vinosum可溶性氢酶的EPR研究

龙敏南1, 苏文金1, Albracht S.P.J2, 张凤章1, 许良树1   

  1. 1. 厦门大学生命科学学院, 细胞生物学与肿瘤细胞工程教育部重点实验室, 厦门 361005;
    2. E.C.SlaterInstitute University of Amsterdam, Amsterdam, the Netherlands
  • 收稿日期:2000-06-14 出版日期:2001-07-24 发布日期:2001-07-24
  • 通讯作者: 龙敏南(1965年出生),男,博士,副教授,从事氢酶生物化学研究.E-mail:longmn@jingxian.xmu.edu.cn E-mail:longmn@jingxian.xmu.edu.cn
  • 基金资助:

    教育部重点科技项目(批准号:99070)

EPR Study of Soluble Hydrogenase from Photosynthetic Bacteria Chromatium vinosum

LONG Min-Nan1, SU Wen-Jin1, Albracht-S. P. J.2, ZHANG Feng-Zhang1, XU Liang-Shu1   

  1. 1. The Key Laboratory of Bioorganic Phosphorus Chemistry, Ministry of Education, Department of Chemistry, School of Life Sciences and Engineering, Tsinghua University, Beijing 100084, China;
    2. E.C.SlaterInstitute University of Amsterdam, Amsterdam, the Netherlands
  • Received:2000-06-14 Online:2001-07-24 Published:2001-07-24

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被引次数

14

摘要： 光合细菌Chromatiumvinosum可溶性氢酶经5次柱层析(DE-23,TSK-DEAE(Ⅰ),UltragelAcA-44,TSK-DEAE(Ⅱ),SuperdexTM75)分离后被纯化365倍,得率为32%,其催化放氢的活性为8.4μmolH2/(min·mgprot).氧化态可溶性氢酶在45K下,产生了Ni(Ⅲ)的典型EPR信号(gx,y,z=2.37,2.16,2.016和gx,y,z=2.30,2.23,2.016);在10K下,没有出现膜结合态氢酶中存在的[3Fe-4S]簇特征EPR信号.可溶性氢酶被H2还原后,Ni(Ⅲ)的特征EPR信号消失,同时出现一个[4Fe-4S]簇的特征EPR信号(gx,y,z=1.88,1.90,2.045).研究结果表明,C.vinosum可溶性氢酶在结构和功能上不同于膜结合态氢酶,是一种新的催化放氢的NiFe-氢酶.

关键词: Chromatiumvinosum, 可溶性氢酶, EPR谱

Abstract: Asoluble hydrogenase(SH) was purified from Chromatium vinosum by five step chromatography(DE-23, TSK-DEAE(I), Ultragel AcA-44, TSK-DEAE(Ⅱ), Superdex TM75) with a specific activity of 8.4 μmol H2/(min·mg prot). The oxidized SHyield two Ni(Ⅲ) EPR(electron paramagnetic resonance) signals( gx,y,z = 2.37, 2.16, 2.016 and gx,y,z = 2.30, 2.23, 2.016 ) at 45 Kwhich occurred in the other NiFe-hydrogenases. However, no[3Fe-4S] cluster EPRsignal was obtained at 10 K. When the SHwas reduced by H2 (over night at 8 ℃), the Ni(Ⅲ) EPRsignals disappeared, and an EPR signal from a reduced[4Fe-4S] cluster appeared( gx,y,z = 1.88, 1.90, 2.045). The results show that the soluble hydrogenase from C.vinosum is a new NiFe hydrogenase which catalyzes H2 production.

Key words: Chromatium vinosum, Soluble hydrogenase, EPR spectra

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