卷枝毛霉△6-脂肪酸脱饱和酶基因的敲除及功能研究

发布时间：2018-04-19 14:41

本文选题：γ-亚麻酸 + 卷枝毛霉　；参考：《食品与生物技术学报》2017年06期

【摘要】：γ-亚麻酸(γ-Linolenic acid,GLA)是人体所必需且具有多种生理功能的一种重要多不饱和脂肪酸。研究表明,卷枝毛霉等GLA生产菌株中△6-脂肪酸脱饱和酶(△6-Fatty acid desaturase,FAD6)是合成γ-亚麻酸的关键和限速酶,然而GLA是否惟一通过△6-脂肪酸脱饱和酶途径合成则目前还没有报道。作者以卷枝毛霉Mucor circinelloides EIM10 sp.为研究对象,以其FAD6基因的5'端启动子和3'端终止子为同源臂,潮霉素B抗性基因为筛选标记,构建了M.circinelloides EIM10 sp.△6-脂肪酸脱饱和酶FAD6的敲除载体p UMCD6-Hm B,转化进入制备好的原生质体中成功地敲除了FAD6基因,发现FAD6基因缺陷菌株GLA质量浓度比野生型对照菌株降低了50%以上,但GLA并没有完全消失,说明卷枝毛霉生物体内还有其他代谢途径可以将碳源转化为GLA。
[Abstract]:纬-linolenic acid (GLAs) is an important polyunsaturated fatty acid which is necessary for human body and has many physiological functions.The results showed that 6-Fatty acid desaturase (FAD6) was the key and rate-limiting enzyme for the synthesis of 纬 -linolenic acid in GLA producing strains, but whether GLA was only synthesized by 6-fatty acid desaturase pathway had not been reported.Mucor circinelloides EIM10 sp.In this study, the 5'terminal promoter and the 3'terminus of FAD6 gene were used as homologous arms. Hygromycin B resistance gene was selected as a screening marker.The knockout vector p UMCD6-Hm B of M.circinelloides EIM10 sp. 6- fatty acid desaturase FAD6 was constructed, and the FAD6 gene was successfully knocked out in the prepared protoplast. It was found that the GLA mass concentration of FAD6 gene deficient strain was more than 50% lower than that of wild type control strain.However, GLA did not completely disappear, suggesting that there are other metabolic pathways in the organism that can convert carbon source into GLAs.
【作者单位】：福建生物工程职业技术学院;福建师范大学工业微生物发酵技术国家地方联合工程研究中心;
【基金】：福建省自然科学基金重大项目(2011H6007);福建省自然科学基金面上项目(2016J01148) 福建省科技计划项目(2014Y2008) 福建省中青年教师教育科研项目(JAT160768)
【分类号】：Q78

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