Gorman D S, Levine R P
The Biological Laboratories, Harvard University, Cambridge, Massachusetts 02138.
Plant Physiol. 1966 Dec;41(10):1643-7. doi: 10.1104/pp.41.10.1643.
Cytochrome 553 and ferredoxin were isolated and purified from acetone powders prepared from intact cells of the wild-type strain of Chlamydomonas reinhardi. Purification was achieved by ion exchange chromatography on DEAE cellulose and gel filtration on Sephadex G-75.Cytochrome 553 could be oxidized with postassium ferricyanide and reduced with sodium ascorbate. The absorption maxima of the cytochrome in reduced form were found to be alpha-band 552.5 nm, beta-band 522.5 nm, and Soret band 416.5 nm. The normal redox potential was +0.37 volt, and the molecular weight was estimated to be 12,000 +/- 2000. The cytochrome closely resembled that of the c-type cytochromes extracted from Euglena and Porphyra. When cultures were harvested after they had entered stationary phase, the cytochrome could be extracted in soluble form, and in some cases a yield of one cytochrome per 1000 chlorophyll molecules could be obtained.Ferredoxin showed absorption maxima at 278, 330, 420 and 460 nm. Thus it resembles the ferredoxin extracted from spinach. The molecular weight of the ferredoxin was estimated to be 15,000 +/- 2000.
从莱茵衣藻野生型菌株的完整细胞制备的丙酮粉中分离并纯化了细胞色素553和铁氧化还原蛋白。通过在DEAE纤维素上进行离子交换色谱和在Sephadex G - 75上进行凝胶过滤实现纯化。细胞色素553可用铁氰化钾氧化，用抗坏血酸钠还原。发现还原形式的细胞色素的吸收最大值为α带552.5nm、β带522.5nm和索雷特带416.5nm。正常氧化还原电位为+0.37伏，分子量估计为12,000±2000。该细胞色素与从眼虫和紫菜中提取的c型细胞色素非常相似。当培养物进入稳定期后收获时，细胞色素可以以可溶形式提取，在某些情况下，每1000个叶绿素分子可获得一个细胞色素的产量。铁氧化还原蛋白在278、330、420和460nm处显示吸收最大值。因此它类似于从菠菜中提取的铁氧化还原蛋白。铁氧化还原蛋白的分子量估计为15,000±2000。
