Wünschiers R, Senger H, Schulz R
Fachbereich Biologie/Botanik, Philipps-Universität, Karl-von-Frisch-Strasse, D-35032, Marburg, Germany.
Biochim Biophys Acta. 2001 Jan 19;1503(3):271-8. doi: 10.1016/s0005-2728(00)00204-8.
The green alga Scenedesmus obliquus is capable of both uptake and production of H(2) after anaerobic adaptation (photoreduction of CO(2) or photohydrogen production). The essential enzyme for H(2)-metabolism is a NiFe-hydrogenase with a [2Fe-2S]-ferredoxin as its natural redox partner. Western blot analysis showed that the hydrogenase is constitutively expressed. The K(m) values were 79.5 microM and 12.5 microM, determined with ferredoxin and H(2), respectively, as electron donor for the hydrogenase. In vitro, NADP(+) was reduced by H(2) in the presence of the hydrogenase, the ferredoxin and a ferredoxin-NADP reductase. From these results and considerations on the stoichiometry we propose that this light-independent electron transfer is part of the photoreduction of CO(2) in vivo. For ATP synthesis, necessary for the photoreduction of CO(2), light-dependent cyclic electron transfer around Photosystem (PS) I accompanies this 'dark reaction'. PS II fluorescence data suggest that (a) in S. obliquus H(2)-reduction might function as the anaerobic counterpart of the O(2)-dependent Mehler reaction, and (b) the presence of either a ferredoxin quinone-reductase or NAD(P)-dehydrogenase (complex I) in S. obliquus chloroplasts.
绿藻斜生栅藻在厌氧适应(二氧化碳的光还原或光产氢)后能够吸收和产生氢气。氢气代谢的关键酶是一种镍铁氢化酶,其天然氧化还原伙伴是一种含[2Fe-2S]的铁氧化还原蛋白。蛋白质免疫印迹分析表明氢化酶是组成型表达的。以铁氧化还原蛋白和氢气分别作为氢化酶的电子供体时,测定的米氏常数(Km)值分别为79.5微摩尔和12.5微摩尔。在体外,在氢化酶、铁氧化还原蛋白和铁氧化还原蛋白-NADP还原酶存在的情况下,氢气可使NADP+还原。基于这些结果以及对化学计量关系的考虑,我们提出这种不依赖光电子传递是体内二氧化碳光还原的一部分。对于二氧化碳光还原所需的ATP合成,围绕光系统(PS)I的依赖光的循环电子传递伴随着这种“暗反应”。PS II荧光数据表明:(a)在斜生栅藻中,氢气还原可能起到了依赖氧气的梅勒反应的厌氧对应作用;(b)斜生栅藻叶绿体中存在铁氧化还原蛋白醌还原酶或NAD(P)脱氢酶(复合体I)。