在工程大肠杆菌中利用 D-葡萄糖和 L-阿拉伯糖高水平生产红景天特征成分玫瑰黄素。

IF 6.8 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Lijun Li , Moshi Liu , Huiping Bi , Tao Liu
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引用次数: 0

摘要

据报道,红景天具有多种生物活性,是世界上广泛使用的一种重要药用植物。然而,野生红景天的濒危状况限制了玫瑰黄素的供应。在这项工作中,我们成功地设计了一种大肠杆菌菌株来高效生产松香黄素,作为一种替代生产方法。首先,从金丝桃中提取肉桂酸:CoA连接酶、肉桂酰-CoA还原酶和枯草芽孢杆菌(Bs-YjiC)的二磷酸尿苷(UDP)-糖基转移酶(UGT)来提高松香在大肠杆菌中的滴度。随后,从 UGT91R 亚家族中鉴定出四种 UGT,可催化松香中松香素的形成,其中来自茄属植物的 SlUGT91R1 活性最高。其次,通过将 SlUGT91R1 和 UDP-阿拉伯糖途径(包括 UDP-葡萄糖脱氢酶、UDP-木糖合成酶和 UDP-木糖 4-epimerase)整合到松香生产染色中,首次在大肠杆菌中实现了松香素的生产,滴度达到 430.5 ± 91.4 mg/L。第三,在大肠杆菌中开发了一条由 L-阿拉伯糖衍生的两步途径,由 L-阿拉伯糖激酶和 UDP-糖焦磷酸酶组成,进一步优化了前体 UDP-阿拉伯糖的供应。此外,利用摇瓶发酵法从 D-葡萄糖和 L-阿拉伯糖中生产出了 1203.7 ± 32.1 mg/L 的玫瑰黄素。最后,通过在 5 升生物反应器中进行饲料批量发酵,生产的玫瑰黄素达到 7539.1 ± 228.7 mg/L。因此,基于微生物的松香黄素生产具有巨大的商业化潜力。这项工作为从 D-葡萄糖和 L-阿拉伯糖生物合成其他含阿拉伯糖单位的有价值天然产物提供了有效的策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
High-level production of Rhodiola rosea characteristic component rosavin from D-glucose and L-arabinose in engineered Escherichia coli

Rosavin is the characteristic component of Rhodiola rosea L., an important medicinal plant used widely in the world that has been reported to possess multiple biological activities. However, the endangered status of wild Rhodiola has limited the supply of rosavin. In this work, we successfully engineered an Escherichia coli strain to efficiently produce rosavin as an alternative production method. Firstly, cinnamate: CoA ligase from Hypericum calycinum, cinnamoyl-CoA reductase from Lolium perenne, and uridine diphosphate (UDP)-glycosyltransferase (UGT) from Bacillus subtilis (Bs-YjiC) were selected to improve the titer of rosin in E. coli. Subsequently, four UGTs from the UGT91R subfamily were identified to catalyze the formation of rosavin from rosin, with SlUGT91R1 from Solanum lycopersicum showing the highest activity level. Secondly, production of rosavin was achieved for the first time in E. coli by incorporating the SlUGT91R1 and UDP-arabinose pathway, including UDP-glucose dehydrogenase, UDP-xylose synthase, and UDP-xylose 4-epimerase, into the rosin-producing stain, and the titer reached 430.5 ± 91.4 mg/L. Thirdly, a two-step pathway derived from L-arabinose, composed of L-arabinokinase and UDP-sugar pyrophosphorylase, was developed in E. coli to further optimize the supply of the precursor UDP-arabinose. Furthermore, 1203.7 ± 32.1 mg/L of rosavin was produced from D-glucose and L-arabinose using shake-flask fermentation. Finally, the production of rosavin reached 7539.1 ± 228.7 mg/L by fed-batch fermentation in a 5-L bioreactor. Thus, the microbe-based production of rosavin shows great potential for commercialization. This work provides an effective strategy for the biosynthesis of other valuable natural products with arabinose-containing units from D-glucose and L-arabinose.

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来源期刊
Metabolic engineering
Metabolic engineering 工程技术-生物工程与应用微生物
CiteScore
15.60
自引率
6.00%
发文量
140
审稿时长
44 days
期刊介绍: Metabolic Engineering (MBE) is a journal that focuses on publishing original research papers on the directed modulation of metabolic pathways for metabolite overproduction or the enhancement of cellular properties. It welcomes papers that describe the engineering of native pathways and the synthesis of heterologous pathways to convert microorganisms into microbial cell factories. The journal covers experimental, computational, and modeling approaches for understanding metabolic pathways and manipulating them through genetic, media, or environmental means. Effective exploration of metabolic pathways necessitates the use of molecular biology and biochemistry methods, as well as engineering techniques for modeling and data analysis. MBE serves as a platform for interdisciplinary research in fields such as biochemistry, molecular biology, applied microbiology, cellular physiology, cellular nutrition in health and disease, and biochemical engineering. The journal publishes various types of papers, including original research papers and review papers. It is indexed and abstracted in databases such as Scopus, Embase, EMBiology, Current Contents - Life Sciences and Clinical Medicine, Science Citation Index, PubMed/Medline, CAS and Biotechnology Citation Index.
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