Development of utilization of lipid containing polyunsaturated fatty acids produced by microbes.

微生物产生的含有多不饱和脂肪酸的脂质的利用开发。

• 批准号: 08455384
• 负责人: SUZUKI Osamu
• 金额: $ 4.99万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08455384/
• 关键词: Polyunsaturated fatty acid; Docosahexaenoic acid; Arachidonic acid; DELTA6 Desaturase; Allergy; Lipid; ドコサヘキサン酸; Schizochytrium sp.; トリアシルグリセロール; 血漿脂質; リン脂質

An isolated fungus Mortierella alliacea YN-15 had a capable of producing 3.9 g/L of arachidonic acid in a flask and 5.0 g/L in a jar fermentor. This strain could metabolize soluble starch as well as glucose. We isolated another arachidonic acid producing microbe Achlya sp.ma-2801 which also contained a significant amout of eicosapentaenoic acid in the cell. Eight stains of microbes producing docosahexaenoic acid were isolated from the sea and one of them, by cultivation in a jar fermentor, appeared to contain high amount of phospholipids consisted of docosahexaenoic acid. A mutant strain MS14-13 that produced higher level of docosahexaenoic acid was isolated from a strain Schizochytrium sp.SR21. The mutant showed higher content of docosahexaenoic acid in total fatty acid than of wild type strain. A gene encoding delta-6 desaturase has been cloned from blue algae and expressed functionally in yeast resulting a successful conversion of exogenious linoleic acid into gamma-linolenic acid.We found, in analyzes of serum fatty acid compositions of allergic patients, RAST value of indivisual patients had a positive correlation with linoleic acid content in their plasma lipid, suggesting the depressed activity of delta-6 desaturase in allergic patients.It is considered that the manipulation of multi-enzyme system of fatty acid desaturation and chain elongation is neccessary for breeding of microbes producing polyunsaturated fatty acids. Isolation of mutants defective in desaturase enzymes and/or cloning of genes coding for such enzymes would be indispensable.

分离到一株被孢霉YN-15，摇瓶发酵生产花生四烯酸3.9g/L，瓶装发酵生产花生四烯酸5.0g/L。该菌株不仅能代谢葡萄糖，还能代谢可溶性淀粉。我们分离到了另一株花生四烯酸产生菌Achlya sp.ma-2801，它在细胞中也含有大量的二十碳五烯酸。从海洋中分离到8株产二十二碳六烯酸的微生物，其中一株在罐式发酵罐中培养，似乎含有大量由二十二碳六烯酸组成的磷脂。从裂殖酵母SR21菌株中分离到一株高产二十二碳六烯酸的突变菌株MS14-13。突变株总脂肪酸中二十二碳六烯酸的含量高于野生型菌株。从蓝藻中克隆了编码Delta-6去饱和酶的基因，并在酵母中进行了功能表达，成功地将外源亚油酸转化为γ-亚麻酸。在对过敏患者血清脂肪酸组成的分析中，我们发现内源性患者的Rast值与其血脂中的亚油酸含量呈正相关，提示过敏患者的Delta-6去饱和酶活性受到抑制。分离有缺陷的去饱和酶突变株和/或克隆编码这种酶的基因将是必不可少的。

项目成果

S.Fujii: "Identification of T cell epitope sequences on an important mite antigen." Clinical and Experimental Allergy. 27. 1086-1094 (1997)

S.Fujii：“鉴定重要螨虫抗原上的 T 细胞表位序列。”

Fujii S: "Identification of T cell epitope Sequences on an impotant mite antigen." Clinical and Experimental Allergy. 27. 1086-1094 (1997)

Fujii S：“重要螨虫抗原上 T 细胞表位序列的鉴定。”

I. Ida, T. Nakahara, et al.: "Improvment of Docosahexaenoic acid Production in a Culturer of Traustochytrium aureum by Medium Optimization" J. Ferment. Bioeng.81 (1). 76-78 (1996)

I. Ida、T. Nakahara 等人：“通过培养基优化提高金黄色葡萄球菌培养器中二十二碳六烯酸的产量”J. Ferment。

O. Hiruta, Y. Kamisaka, et al.: "γ-Linoleic Acid Produotion by a Low Temperature Resistant Mutant of Nortierella oamaniana." J. Ferment. Bioeng.82 (1). 119-123 (1996)

O. Hiruta、Y. Kamisaka 等人：“Nortierella oamaniana 耐低温突变体生产 γ-亚油酸”。J. Bioeng.82 (1)。

Fujii S: "Identification of T cell epitope sequences on an imptant mite antigen." Clinical and Experimental Allergy. 27. 1086-1094 (1997)

Fujii S：“识别植入性螨虫抗原上的 T 细胞表位序列。”