生产植物三萜:亚洲酸、马来酸和arjunolic酸的解脂耶氏菌工程

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Jonathan Asmund Arnesen , Arian Belmonte Del Ama , Sidharth Jayachandran , Jonathan Dahlin , Daniela Rago , Aaron John Christian Andersen , Irina Borodina
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引用次数: 7

摘要

几种植物三萜具有宝贵的药用特性,但由于从植物中提取会给自然资源造成负担,而且产量和纯度都很低,因此它们的生产和使用受到限制。在这里,我们设计了产油酵母解脂耶氏酵母,通过发酵生产三种有价值的植物三萜(亚洲酸、马来酸和arjunolic酸)。首先,我们建立了重组生产的前体,熊果酸和齐墩果酸,通过表达植物酶的游离或融合版本的脂肪瘤菌株之前优化为角鲨烯生产。工程菌株产生高达11.6 mg/g熊果酸或10.2 mg/g齐墩果酸。通过表达积雪草细胞色素P450单加氧酶CaCYP716C11p、CaCYP714E19p和CaCYP716E41p,延长了熊果酸的生物合成途径,产生了微量的积雪酸和0.12 mg/g DCW的合成酸。在齐墩果酸产生菌株中表达相同的亚洲木曲细胞色素P450,产生齐墩果酸三萜。在齐墩果酸产生菌株中,CaCYP716C11p的表达产生8.9 mg/g DCW的山楂酸。进一步表达经过密码子优化的CaCYP714E19p可产生4.4 mg/g DCW的arjunolic acid。最后,通过将CaCYP714E19p的n -末端结构域与Kalopanax septemlobus细胞色素P450的n -末端结构域交换,将arjunolic acid的产量提高到9.1 mg/g DCW。总之,我们已经证明了在微生物细胞工厂生产亚细亚酸、马来酸和arjunolic酸。在工业化生产这些分子之前,菌株和发酵工艺需要进一步改进。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Engineering of Yarrowia lipolytica for the production of plant triterpenoids: Asiatic, madecassic, and arjunolic acids

Several plant triterpenoids have valuable pharmaceutical properties, but their production and usage is limited since extraction from plants can burden natural resources, and result in low yields and purity. Here, we engineered oleaginous yeast Yarrowia lipolytica to produce three valuable plant triterpenoids (asiatic, madecassic, and arjunolic acids) by fermentation. First, we established the recombinant production of precursors, ursolic and oleanolic acids, by expressing plant enzymes in free or fused versions in a Y. lipolytica strain previously optimized for squalene production. Engineered strains produced up to 11.6 mg/g DCW ursolic acid or 10.2 mg/g DCW oleanolic acid. The biosynthetic pathway from ursolic acid was extended by expressing the Centella asiatica cytochrome P450 monoxygenases CaCYP716C11p, CaCYP714E19p, and CaCYP716E41p, resulting in the production of trace amounts of asiatic acid and 0.12 mg/g DCW madecassic acid. Expressing the same C. asiatica cytochromes P450 in oleanolic acid-producing strain resulted in the production of oleanane triterpenoids. Expression of CaCYP716C11p in the oleanolic acid-producing strain yielded 8.9 mg/g DCW maslinic acid. Further expression of a codon-optimized CaCYP714E19p resulted in 4.4 mg/g DCW arjunolic acid. Lastly, arjunolic acid production was increased to 9.1 mg/g DCW by swapping the N-terminal domain of CaCYP714E19p with the N-terminal domain from a Kalopanax septemlobus cytochrome P450. In summary, we have demonstrated the production of asiatic, madecassic, and arjunolic acids in a microbial cell factory. The strains and fermentation processes need to be further improved before the production of these molecules by fermentation can be industrialized.

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来源期刊
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
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