Zhang Yue, Zhang Sufang, Chu Yadong, Zhang Qi, Zhou Renhui, Yu Di, Wang Shuang, Lyu Liting, Xu Guowang, Zhao Zongbao Kent
Laboratory of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China.
University of Chinese Academy of Sciences, Beijing, China.
Front Bioeng Biotechnol. 2022 Oct 26;10:1034972. doi: 10.3389/fbioe.2022.1034972. eCollection 2022.
The basidiomycetous yeast () is an excellent producer for neutral lipids, including triacylglycerols (TAG). Partially because genetic tools for this yeast were less developed, limited efforts were shown to explore its capacity for the production of higher-value lipids such as diacylglycerols (DAG). Here, four genes linked to the interconversion between DAG and TAG were manipulated to promote the production of DAG and free fatty acids (FFA). Among them, three TAG synthesis-related genes, , and , were down-regulated successively the RNA interference technology, and an endogenous TAG lipase encoded by was fused with and over-expressed to convert TAG into DAG and FFA. Results showed that those engineered strains grew normally under nutrient-rich conditions but notably slower than the parental strain NP11 in the lipid production stage. When cultivated in nitrogen-limited media, engineered strains were able to produce total lipids with improved contents of DAG and FFA by up to two-fold and three-fold, respectively. Further correlation analysis between lipid composition and cell density indicated that the formation of TAG correlated positively with cell growth; however, other lipids including DAG did negatively. This study offered valuable information and strains to engineer for advanced production of fatty acid derivatives.
担子菌酵母()是中性脂质(包括三酰甘油(TAG))的优秀生产者。部分原因是该酵母的遗传工具开发较少,因此探索其生产更高价值脂质(如二酰甘油(DAG))的能力的努力有限。在此,对与DAG和TAG相互转化相关的四个基因进行了操作,以促进DAG和游离脂肪酸(FFA)的生产。其中,三个与TAG合成相关的基因,和,通过RNA干扰技术依次下调,并且由编码的内源性TAG脂肪酶与融合并过表达,以将TAG转化为DAG和FFA。结果表明,那些工程化的菌株在营养丰富的条件下正常生长,但在脂质生产阶段明显比亲本菌株NP11生长得慢。当在氮限制培养基中培养时,工程化菌株能够生产总脂质,其中DAG和FFA的含量分别提高了两倍和三倍。脂质组成与细胞密度之间的进一步相关性分析表明,TAG的形成与细胞生长呈正相关;然而,包括DAG在内的其他脂质则呈负相关。这项研究为工程化生产脂肪酸衍生物提供了有价值的信息和菌株。