Bury N R, Codd G A, Wendelaaar Bonga S E, Flik G
Department of Animal Physiology, University of Nijmegen, The Netherlands.
J Exp Biol. 1998 Jan;201(Pt 1):81-9. doi: 10.1242/jeb.201.1.81.
Fatty acids from two strains of the cyanobacterium Microcystis aeruginosa, PCC 7820 (a strain that produces the hepatotoxin microcystin-LR, MC-LR) and CYA 43 (a strain that produces only small quantities of MC-LR), were extracted, partially characterised and tested for their inhibitory effect on the K+-dependent p-nitrophenol phosphatase (pNPPase) activity of tilapia (Oreochromis mossambicus) gill basolateral membrane. Thin-layer chromatography of the lipids from dichloromethane:methanol extracts of M. aeruginosa PCC 7820 and CYA 43, using diethylether:isopropanol:formic acid (100:4.5:2.5) as solvent, yielded five inhibitory products from M. aeruginosa 7820 and six from M. aeruginosa CYA 43. None of these products could be related to MC-LR. The inhibitory behaviour of the products mimics that of a slow, tight-binding inhibitor. The inhibitory activity is removed by incubation of extracts with fatty-acid-free bovine serum albumin (FAF-BSA). However, FAF-BSA only partially reversed the inhibition of K+-dependent pNPPase on fish gills pre-exposed to the extracted products. We conclude that M. aeruginosa strains PCC 7820 and CYA 43 produce fatty acids with potent inhibitory effects on K+-dependent pNPPase. The release of these products following lysis of cyanobacterial blooms may help to explain fish kills through a disturbance of gill functioning.
从两种铜绿微囊藻菌株中提取脂肪酸,这两种菌株分别是PCC 7820(一种产生肝毒素微囊藻毒素-LR,即MC-LR的菌株)和CYA 43(一种仅产生少量MC-LR的菌株)。对提取的脂肪酸进行了部分特性鉴定,并测试了它们对罗非鱼(莫桑比克罗非鱼)鳃基底外侧膜上钾离子依赖性对硝基苯酚磷酸酶(pNPPase)活性的抑制作用。以二氯甲烷:甲醇提取物对铜绿微囊藻PCC 7820和CYA 43进行脂质的薄层色谱分析,使用乙醚:异丙醇:甲酸(100:4.5:2.5)作为溶剂,从铜绿微囊藻7820中得到了5种抑制产物,从铜绿微囊藻CYA 43中得到了6种抑制产物。这些产物均与MC-LR无关。这些产物的抑制行为类似于一种缓慢、紧密结合的抑制剂。通过将提取物与无脂肪酸牛血清白蛋白(FAF-BSA)孵育可消除抑制活性。然而,FAF-BSA只能部分逆转预先暴露于提取产物的鱼鳃上钾离子依赖性pNPPase的抑制作用。我们得出结论,铜绿微囊藻菌株PCC 7820和CYA 43产生的脂肪酸对钾离子依赖性pNPPase具有强效抑制作用。蓝藻水华裂解后这些产物的释放可能有助于解释通过鳃功能紊乱导致的鱼类死亡现象。