发状念珠藻(发菜)细胞质膜的分离与性质分析

使用一种新方法首次从野生发菜(Nostoc flagelliforme Born.et Flah.)中分离得到细胞质膜并对其性质进行了分析,该方法的主要特点为联合使用细胞破碎仪和毛地黄皂甙对发菜细胞进行破碎.经过细胞破碎仪处理两次(80MPa)后,样品(20mg干重/mL)中的细胞可被毛地黄皂甙(3mg/mL)有效破碎,细胞质膜即可通过蔗糖密度梯度离心得以分离.纯化后的质膜,其吸收光谱中类胡萝卜素的3个吸收峰分别位于458、487和524 nm,另外一种叶绿素前体在673 nm处有少量吸收,质膜的荧光发射来自该叶绿素前体.通过变性电泳对其进行多肽组成分析,可分辨出30多条多肽,其中分子量为80、28、19和17 kD的多肽含量最高.其膜脂主要包含4种成分:单半乳糖甘油二酯(62.4%)、双半乳糖甘油二酯(18.9%)、硫代异鼠李糖甘油二酯(16.7%)和磷酯酰甘油(2.0%).膜脂酯酰基连接有棕榈酸(16:0)、十六碳烯酸(16:19])、硬脂酸(18:0)、油酸(18:19])、亚油酸(18:29,12])和亚麻酸(18:39,12,15])等六种脂肪酸,其中十六碳烯酸和亚麻酸为主要成分,分别占总脂肪酸含量的32.3%和34.4%.质膜中高含量的亚麻酸可能是发菜具有极强抗旱能力的一个重要因素.

Isolation and Characterization of the Cytoplasmic Membrane from the Terrestrial Cyanobacterium-Nostoc flagelliforme

Cytoplasmic membrane of Nostoc flagelliforme Born. et Flah. was isolated for the first time with a new method, the unique feature of which is the combined use of French pressure cell and digitonin to disrupt cells. After passed twice through French pressure cell (at 80 MPa), cells in sample (20 mg of dry weight/mL) were disrupted effectively by digitonin (3 mg/mL), and then the cytoplasmic membrane was isolated by density gradient centrifugation. The membrane contained carotenoids with absorption peaks at 458, 487 and 524 nm and a precursor of chlorophyll a with a minor peak at 673 nm. The fluorescence emission peaks of the membrane were emitted from the precursor of chlorophyll a. More than 30 polypeptides were detected in the membrane, in which the most obvious corresponded to the polypeptides with molecular mass of 80, 28, 19 and 17 kD. The membrane contained four types of glycerolipids: MGDG (62.4%), DGDG (18.9%), SQDG (16.7%) and PG (2.0%). 16:0, 16:1 9], 18:0, 18:1 9], 18:2 9, 12] and 18:3 9, 12, 15] fatty acids were determined in the membrane, in which 16:1 and 18:3 fatty acids were the main components, representing 32.3% and 34.4% of the total fatty acids respectively. High proportion of 18:3 fatty acid in the cytoplasmic membrane may be an important factor of N. flagelliforme in its remarkable drought-tolerant ability.