State Key Laboratory of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, People's Republic of China.
State Key Laboratory of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, People's Republic of China.
Bioresour Technol. 2015 Feb;177:134-40. doi: 10.1016/j.biortech.2014.11.051. Epub 2014 Nov 25.
To obtain mutant strains with higher arachidonic acid (ARA) yields, the oleaginous fungus Mortierella alpina was mutated using atmospheric and room temperature plasma (ARTP) coupled with diethyl sulfate (DES). A visual compound filter operation was used in which a screening medium was supplemented with cerulenin, an inhibitor of fatty acid synthase (FAS), and triphenyltetrazolium chloride (TTC). The mutant strain D20 with an ARA production of 5.09 g/L, a 40.61% increase over the original strain (3.62 g/L), was isolated. The relative ARA content increased from 38.99% to 45.64% of total fatty acids. After optimizing fermentation conditions, the maximum ARA yield (6.82 g/L) for strain D20 was obtained in shake flasks. This work provides an appropriate strategy for obtaining high ARA-yield strains by conventional random mutation methods with an efficient screening assay.
为了获得具有更高花生四烯酸（ARA）产量的突变株，本研究采用常压室温等离子体（ARTP）与硫酸二乙酯（DES）相结合的方法对产油真菌枝顶孢霉进行诱变。采用可视化复合筛选操作，即在筛选培养基中添加脂肪酸合酶（FAS）抑制剂苍术苷和氯化三苯基四氮唑（TTC）。分离得到 ARA 产量为 5.09 g/L 的突变株 D20，比原始菌株（3.62 g/L）提高了 40.61%。ARA 的相对含量从总脂肪酸的 38.99%增加到 45.64%。在优化发酵条件后，摇瓶中突变株 D20 的最大 ARA 产量（6.82 g/L）。这项工作为利用常规随机突变方法并结合有效的筛选方法获得高产 ARA 菌株提供了一种合适的策略。
