基于代谢控制育种技术选育深黄被孢霉γ-亚麻酸高产菌株  

Screening of Mortierella isabellina mutants for high-yield γ-linolenic acid by metabolic control breeding technique

作　　者：郝冉[1] 楚乐乐 胡申才[1] 

机构地区：[1]武汉轻工大学生物与制药工程学院,湖北武汉430023

出　　处：《中国酿造》2016年第4期51-55,共5页China Brewing

基　　金：湖北省科技厅自然基金项目(2013CFB326)

摘　　要：为了获得深黄被孢霉合成γ-亚麻酸(GLA)的高产菌株,该研究采用了紫外照射和氮离子注入技术2种诱变方法建立突变体库。根据γ-亚麻酸代谢合成途径中的部分关键因子,设计并建立抗丙二酸和红四氮唑(TTC)筛选模型,旨在提高菌体生物量和不饱和脂肪酸含量。结果显示,采用丙二酸质量浓度为2 g/L的平板可筛选得到高产油脂菌株;在发酵培养的第54小时收集菌体,用0.9%TTC染色4 h,可作为TTC法的最佳染色条件。经过两轮不同方法的诱变筛选后,最终得到了一株具良好遗传稳定性的突变株I-0281,其GLA产量为889.80 mg/L,比原始菌株提高了60.27%。In order to obtain Mortierella isabellina mutants with high γ-linolenic acid yield, the mutant library was constructed by UV and N^＋ ions implantation techniques, respectively. Then according to some key enzymes involved in the metabolic synthesis pathway of γ-linolenic acid, the anti-malonic acid model and triphenyltetrazolium chloride （TTC） staining model were designed and established for increasing biomass and unsaturated fatty acid content. The results showed that the high-yield strains could be screened by resistant plates containing 2 g/L malonic acid. The optimal staining conditions pertaining to TTC screening model were determined as follows： collecting samples after fermentation for 54 h, then stained by 0.9% TTC for 4 h. Through two rounds screening via the two kinds of screening model, the mutant strain I-0281, which had good genetic stability was obtained. The production of γ-linolenic acid was up to 889.80 mg/L, which was 60.27% higher than that from the original strain.

关 键 词：Γ-亚麻酸 菌种选育 筛选模型 抗丙二酸法 红四氮唑法 

分 类 号：Q93[生物学—微生物学]