Characterization and role of cis/trans isomerase of unsaturated fatty acids in bacteria.

细菌中不饱和脂肪酸顺/反异构酶的表征和作用。

• 批准号: 01540557
• 负责人: OKUYAMA Hidetoshi
• 金额: $ 1.28万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01540557/
• 关键词: Psychrotroph; Pseudomonas; Psychrophile; Vibrio; cis; trans isomerase; trans-Unsaturated fatty acid; Pseudomonas; シス・トランス異性化酵素; 脂肪酸; 低温適応; Vibrio ABE-1株; 不飽和脂肪酸異性化酵素

Cell-free extract of PseudoNonas sp. strain E-3 catalyzed the conversion of 9cis-hexadecenoib acid[16 : 1(9c)]in a free acid and 16 : 1(9c)esterified to both the sn-1 and 2 positions of phosphatidylethanolamine(PE)to 9-trans-hexadecenoic acid[16 : 1(9t)]. The cell-free extract wa!j separated into the soluble(cytosolic)and membrane fractions by the ultracentrifugation at 105, 000 x g for 60 min. The cytosol fraction catalyzed the isomerization of the free 16 : 1(9c). On the other hand, it isomerized 16 : 1(9c)esterified to PE in the presence of the membrane fraction. One cytosolic component(component I)that catalyze the isomerization of free 16 : 1(9c)and the other(component II)that is needed for the isomerization of esterified 16 : 1(9c)were completely separable one another with a high-performance liquid chromatographic system equipped Kith a gel filtration column. These results suggest that Psedomonas sp. strain E-3 contains at least two types of "Cis/trans isometases.Component I was purified by column chromatography with DEAE-Toyopearl and ButyToyopearl. Purified enzyme was separated into two components by gel-filtration column chromatography. These two components ' were essentially needed for isomeriation of 16 : 1(9c)to 16 : 1(9t). The enzyme has a temperature optimum at ^oC and maintained significant activities at subzero temperatures.

假Nonas sp.菌株E-3的无细胞提取物催化游离酸中的9顺式-十六碳烯酸[16：1（9 c）]和与磷脂酰乙醇胺（PE）的sn-1和2位酯化的16：1（9 c）转化为9-反式-十六碳烯酸[16：1（9 t）]。无细胞提取物哇！通过在105，000 x g下超离心60 min，分离成可溶性（胞质）和膜级分。胞质级分催化游离16：1（9 c）的异构化。另一方面，它异构化16：1（9 c）酯化PE的存在下的膜馏分。催化游离16：1（9 c）异构化的一种胞质组分（组分I）和酯化16：1（9 c）异构化所需的另一种组分（组分II）可以用配备有凝胶过滤柱的高效液相色谱系统完全分离。这些结果表明，假单胞菌E-3菌株至少含有两种类型的“顺/反"异构酶。纯化后的酶经凝胶过滤柱层析分离为两种组分。这两种组分是16：1（9 c）异构化为16：1（9 t）所必需的。该酶的最适温度为1 ℃，在零下温度下仍能保持显著的活性。

项目成果

N.Morita,H.Hayami,H.Okuyama,N.Goto,N.Okajima,H.Higahi & N.Murata: "Both the anaerobic pathway and aerobic desaturation are involved in synthesis of unsaturated fatty acids in Vibrio sp.Strain ABE-1." FEBS Letters. 297. 9-12 (1992)

森田N、速水H、奥山H、后藤N、冈岛N、东日H.

H.Okuyama,N.Morita & K.Kogami: "The occurrence of octadecapentaenoic acids in the lipids of a cold stenothermic alga,prymnesiophyte strain B" J.Phycol.

H·奥山、N·森田

N. Morita, H. Hayashi, H. Okuyama, N. Gotoh, N. Okajima, H. Higashi, and N. Murata: "Both the anaerobic pathway and aerobic desaturation are involved in synthesis of unsaturated fatty acids in Vibrio sp. strain ABE-1." FEBS Letters. 297. 9-12 (1992)

N. Morita、H. Hayashi、H. Okuyama、N. Gotoh、N. Okajima、H. Higashi 和 N. Murata：“厌氧途径和有氧去饱和都参与弧菌菌株不饱和脂肪酸的合成”

R.Kawazoe and H.Okuyama: "Pufification and characterization of △9ーhexadecenoic acid cis/trans isomerase from Pseudomonas sp.strain Eー3" J.Biol.Chem.

R.Kawazoe 和 H.Okuyama：“来自假单胞菌菌株 E-3 的 △9-十六碳烯酸顺/反异构酶的纯化和表征”J.Biol.Chem。

N.Morita,他: "Synthesis in vitro of very long chain fotty acids by fatty acid synthetase from Vibrio sp.strain ABEー1" J.Bacteri ology.

N.Morita 等人：“通过来自弧菌菌株 ABE-1 的脂肪酸合成酶体外合成极长链脂肪酸”J.Bacteriology。