雅罗维亚
聚酮
代谢工程
异源表达
生物
异源的
酵母
生物合成
生物化学
清脆的
波姆裂殖酵母
基因
基因组
酿酒酵母
重组DNA
作者
Mihaela Bejenari,Eva Mie Lang Spedtsberg,Jesper Mosolff Mathiesen,Alexandra Claire Jeppesen,Lucia Cernat,Ariane Toussaint,Cristina Apostol,Victor Stoianov,Tobias Bruun Pedersen,Mikkel Rank Nielsen,Jens Laurids Sørensen
标识
DOI:10.3389/ffunb.2024.1327777
摘要
Fungal polyketides are a large group of secondary metabolites, valuable due to their diverse spectrum of pharmacological activities. Polyketide biosynthesis in filamentous fungi presents some challenges: small yield and low-purity titers. To tackle these issues, we switched to the yeast Yarrowia lipolytica , an easily cultivable heterologous host. As an oleaginous yeast, Y. lipolytica displays a high flux of acetyl- and malonyl-CoA precursors used in lipid synthesis. Likewise, acetyl- and malonyl-CoA are the building blocks of many natural polyketides, and we explored the possibility of redirecting this flux toward polyketide production. Despite its promising prospect, Y. lipolytica has so far only been used for heterologous expression of simple type III polyketide synthases (PKSs) from plants. Therefore, we decided to evaluate the potential of Y. lipolytica by targeting the more complex fungal polyketides synthesized by type I PKSs. We employed a CRISPR-Cas9-mediated genome editing method to achieve markerless gene integration of the genes responsible for bostrycoidin biosynthesis in Fusarium solani ( fsr1 , fsr2 , and fsr3 ) and 6-methylsalicylic acid (6-MSA) biosynthesis in Aspergillus hancockii (6MSAS). Moreover, we attempted titer optimization through metabolic engineering by overexpressing two enzymes, TGL4 and AOX2, involved in lipid β-oxidation, but we did not observe an effect on polyketide production. With maximum titers of 403 mg/L 6-MSA and 35 mg/L bostrycoidin, the latter being substantially higher than our previous results in Saccharomyces cerevisiae (2.2 mg/L), this work demonstrates the potential of Y. lipolytica as a platform for heterologous production of complex fungal polyketides.
科研通智能强力驱动
Strongly Powered by AbleSci AI