Preparation of chlorophyll a, chlorophyll b and bacteriochlorophyll a by means of column chromatography with diethylaminoethylcellulose.
Preparation of chlorophyll a, chlorophyll b and bacteriochlorophyll a by means of column chromatography with diethylaminoethylcellulose.
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二乙氨基乙基纤维素柱色谱法制备叶绿素a、叶绿素b和细菌叶绿素a。
DOI：
10.1016/0005-2728(78)90099-3
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发表时间：
1978
期刊：
影响因子：
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通讯作者：
N. Murata
中科院分区：
文献类型：
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作者：
N. Sato;N. Murata
用SephadexLH-20和二乙氨基乙基纤维素柱层析法分离出叶绿素a、叶绿素b和细菌叶绿素a。该方法在约3小时内提供叶绿素的纯化制剂。为了制备叶绿素，使用蓝绿藻或红藻作为起始材料。用90%丙酮水溶液从多变鱼腥藻、构巢线囊藻和细单歧藻的细胞中提取叶绿素，用90%甲醇水溶液从条斑紫菜的菌体中提取叶绿素。根据Iriyama等人[6]的方法，通过向提取物中加入二氧六环和水，以沉淀形式收集叶绿素。将叶绿素粗品上SephadexLH-20柱，用氯仿作洗脱剂，再上DEAE-纤维素柱，用氯仿/甲醇（49：1，v/v）作洗脱剂。用薄层色谱分析表明，叶绿素制备物不含可检测的污染物。以类似的方法从紫色光合细菌、球形红球藻和葡萄色菌中制备细菌叶绿素a。叶绿素a和b的混合物以与从蓝绿藻制备叶绿素a所述相同的方式获得。为了从叶绿素中分离叶绿素b，将混合物施加到二乙基氨基乙基纤维素柱上，该柱用己烷/2-丙醇混合物（5：2，v/v）展开。
Chlorophylla, chlorophyllband bacteriochlorophyllawere prepared by means of column chromatography with Sephadex LH-20 and diethylaminoethylcellulose. This method provides purified preparations of chlorophylls in about 3 h.To prepare chlorophylla, blue-green or red algae were used as the starting material. Chlorophyllawas extracted with 90% aqueous acetone from cells of blue-green algae,Anabaena variabilis, Anacystis nidulansandTolypothrix tenuis, and with 90% aqueous methanol from thalli of a red alga,Porphyra yezoensis. Chlorophyllawas collected as precipitates by adding dioxane and water to the extract according to the method of Iriyama et al. [6]. The crude chlorophyllapreparation was applied to a Sephadex LH-20 column with chloroform as the eluent and then to a DEAE-cellulose column with a chloroform/methanol mixture (49 : 1, v/v) as the eluent. Analysis with thin layer chromatography revealed that the chlorophyllapreparation contained no detectable contaminants.Bacteriochlorophyllawas prepared in a similar manner from purple photosynthetic bacteria,Rhodopseudomonas spheroidesandChromatium vinosum.In order to prepare chlorophyllb, chloroplasts of spinach leaves were used as the starting material. A mixture of chlorophyllsaandbwas obtained in the same way as described for the preparation of chlorophyllafrom the blue-green algae. To separate chlorophyllbfrom chlorophylla, the mixture was applied to a diethylaminoethylcellulose column which was developed with a hexane/2-propanol mixture (5 : 2, v/v).
东京大学 - 微生物学
DOI：
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