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来自……的角鲨烯环氧酶的功能表征

Functional Characterization of Squalene Epoxidases from .

作者信息

Zhao Huan, Song Ze, Liu Xuan, Gong Shukun, Tang Qi, Liu Changli, Zhang Yifeng, Zhang Xianan, Gao Haiyun, Gao Wei, Hu Yating, Huang Luqi

机构信息

School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.

College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China.

出版信息

Plants (Basel). 2025 Jun 6;14(12):1740. doi: 10.3390/plants14121740.

DOI:10.3390/plants14121740
PMID:40573728
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12196517/
Abstract

The medicinal plant produces sweet-tasting cucurbitane-type mogrosides from the atypical triterpenoid precursor 2,3,22,23-dioxidosqualene (SDO), rather than the conventional 2,3-oxidosqualene (SQO). However, SDO formation in mogroside biosynthesis remains unclear. Here, we systematically characterized two squalene epoxidases (SgSQE1/2) through phylogenetic analysis, heterologous expression, subcellular localization, qRT-PCR, and alanine scanning studies. Both SQE1 and SQE2 exhibited squalene epoxidase activity, with SQE2 catalyzing SDO formation in yeast. We identified two critical catalytic residues governing epoxidation efficiency through mutagenesis. Both SQEs were localized in the ER, while expression profiling revealed a similar trend between expression and mogroside accumulation in fruits. In our study, we developed a genomically engineered strategy for heterologous SQE characterization. These results lay the foundation for the SQE catalytic reaction involved in mogroside biosynthesis, and provide gene resources and a feasible approach for triterpene metabolic engineering.

摘要

这种药用植物从非典型三萜前体2,3,22,23-二氧化角鲨烯（SDO）而非传统的2,3-氧化角鲨烯（SQO）中产生甜味的葫芦烷型罗汉果苷。然而，罗汉果苷生物合成中SDO的形成仍不清楚。在此，我们通过系统发育分析、异源表达、亚细胞定位、qRT-PCR和丙氨酸扫描研究，对两种角鲨烯环氧酶（SgSQE1/2）进行了系统表征。SQE1和SQE2均表现出角鲨烯环氧酶活性，其中SQE2在酵母中催化SDO的形成。我们通过诱变确定了两个决定环氧化效率的关键催化残基。两种SQE均定位于内质网，而表达谱分析显示果实中表达与罗汉果苷积累之间存在相似趋势。在我们的研究中，我们开发了一种用于异源SQE表征的基因组工程策略。这些结果为罗汉果苷生物合成中涉及的SQE催化反应奠定了基础，并为三萜代谢工程提供了基因资源和可行方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/a1683ed43bb8/plants-14-01740-g013.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/dd32ae44a06d/plants-14-01740-g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/c9904e48b29c/plants-14-01740-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/57e862719bbe/plants-14-01740-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/6404e5680a5b/plants-14-01740-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/71be8e534ab7/plants-14-01740-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/b8380dec1b85/plants-14-01740-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/3c90c51959da/plants-14-01740-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/595e984064ec/plants-14-01740-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/158a54afb598/plants-14-01740-g011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/3bdebe917749/plants-14-01740-g012.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/a1683ed43bb8/plants-14-01740-g013.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/dd32ae44a06d/plants-14-01740-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/7281d647f79f/plants-14-01740-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/46a3685a43aa/plants-14-01740-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/c9904e48b29c/plants-14-01740-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/57e862719bbe/plants-14-01740-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/6404e5680a5b/plants-14-01740-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/71be8e534ab7/plants-14-01740-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/b8380dec1b85/plants-14-01740-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/3c90c51959da/plants-14-01740-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/595e984064ec/plants-14-01740-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/158a54afb598/plants-14-01740-g011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/3bdebe917749/plants-14-01740-g012.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/acb9/12196517/a1683ed43bb8/plants-14-01740-g013.jpg

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本文引用的文献

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Nanoplastics Chronic Toxicity in Mice: Disturbing the Homeostasis of Tryptophan Metabolism in Gut-Lung-Microbiota Axis.纳米塑料对小鼠的慢性毒性：扰乱肠道-肺-微生物群轴中色氨酸代谢的稳态
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Mogroside V protects against acetaminophen-induced liver injury by reducing reactive oxygen species and c-jun-N-terminal kinase activation in mice.罗汉果甜苷V通过减少小鼠体内的活性氧和c-Jun氨基末端激酶激活来预防对乙酰氨基酚诱导的肝损伤。
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Mogroside V ameliorates astrocyte inflammation induced by cerebral ischemia through suppressing TLR4/TRADD pathway.
罗汉果甜苷V通过抑制TLR4/TRADD信号通路减轻脑缺血诱导的星形胶质细胞炎症。
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De novo biosynthesis of mogroside V by multiplexed engineered yeasts.通过多重工程酵母从头生物合成罗汉果甜苷V
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Overproduction of Cucurbitadienol through Modular Metabolic Engineering and Fermentation Optimization in .通过模块化代谢工程和发酵优化在……中过量生产葫芦二烯醇
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Reengineering the Substrate Tunnel to Enhance the Catalytic Efficiency of Squalene Epoxidase.重新设计底物隧道以提高角鲨烯环氧化酶的催化效率。
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Structural insights into the catalytic selectivity of glycosyltransferase SgUGT94-289-3 towards mogrosides.糖苷转移酶 SgUGT94-289-3 对罗汉果苷的催化选择性的结构见解。
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Mogroside Alleviates Diabetes Mellitus and Modulates Intestinal Microflora in Type 2 Diabetic Mice.蜜瓜苷可缓解 2 型糖尿病小鼠的糖尿病症状并调节其肠道菌群。
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Mogroside V reduced the excessive endoplasmic reticulum stress and mitigated the Ulcerative colitis induced by dextran sulfate sodium in mice.罗汉果苷 V 减轻了过度的内质网应激，缓解了葡聚糖硫酸钠诱导的小鼠溃疡性结肠炎。
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