Sakuradani Eiji, Hirano Yuriko, Kamada Nozomu, Nojiri Masutoshi, Ogawa Jun, Shimizu Sakayu
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto, 606-8502, Japan.
Appl Microbiol Biotechnol. 2004 Dec;66(3):243-8. doi: 10.1007/s00253-004-1682-7.
Five mutants were obtained, Y11, Y135, Y164, Y180 and Y61, capable of accumulating higher amounts of arachidonic acid (AA) than Mortierella alpina 1S-4, an industrial strain for the production of AA-rich triacylglycerol (TG). This is thought to be due to low or no activity of n-3 desaturation with conversion of AA to eicosapentaenoic acid, which functions at a cultural temperature below 20 degrees C. In small-scale cultivation under optimum conditions, Y11 and Y61 respectively accumulated 4.97 mg/ml and 4.11 mg/ml of AA, using a high concentration of glucose at 20 degrees C, compared with 3.74 mg/ml for M. alpina 1S-4. In a 5-1 jar fermentor, the AA content in Y11 and Y61 kept increasing during cultivation, with consumption of the glucose in the medium; and this reached 1.48 mg/ml and 1.77 mg/ml (118 mg/g, 120 mg/g of dry mycelia) at day 10, respectively, compared with 0.95 mg/ml (86 mg/g of dry mycelia) for M. alpina 1S-4. From the results of lipid analysis, the TG contents of Y11 and Y61 in the major lipids were significantly higher than that of M. alpina 1S-4; and the AA percentages in TG of Y11 and Y61 were also higher. Both Y11 and Y61 are potential producers of TG rich in AA.
获得了五个突变体,即Y11、Y135、Y164、Y180和Y61,它们能够比高山被孢霉1S-4积累更多的花生四烯酸(AA),高山被孢霉1S-4是一种用于生产富含AA的三酰甘油(TG)的工业菌株。据认为,这是由于n-3去饱和活性低或无活性,使得AA无法转化为二十碳五烯酸,而n-3去饱和在20℃以下的培养温度下起作用。在最佳条件下进行小规模培养时,Y11和Y61在20℃下使用高浓度葡萄糖分别积累了4.97mg/ml和4.11mg/ml的AA,相比之下,高山被孢霉1S-4积累的量为3.74mg/ml。在5升罐式发酵罐中,Y11和Y61中的AA含量在培养过程中随着培养基中葡萄糖的消耗而持续增加;在第10天分别达到1.48mg/ml和1.77mg/ml(干菌丝体为118mg/g、120mg/g),相比之下,高山被孢霉1S-4为0.95mg/ml(干菌丝体为86mg/g)。从脂质分析结果来看,Y11和Y61主要脂质中的TG含量显著高于高山被孢霉1S-4;Y11和Y61的TG中AA的百分比也更高。Y11和Y61都是富含AA的TG的潜在生产菌株。