Biosynthesis of pentacyclic triterpenoid saponins in microbial cell factories

doi:10.12211/2096-8280.2020-082

Abstract

Abstract:

Pentacyclic triterpenoid saponins are important plant natural products, which have five rings composed of six isoprene units. They are widely used in medicine, functional food, health care products, cosmetics and other fields due to their pharmacological and physiological activities. At present, pentacyclic triterpenoid saponins are derived mainly from direct extraction from plants, which has many disadvantages such as the long growth period of plants, competition for farmland, impact on ecological environment and low extraction yield. It is also difficult to obtain pentacyclic triterpenoid saponins through chemical synthesis because of the complexity of their structures, and in the meantime the process using organic reagents and performed under harsh reaction conditions is not environmentally friendly. In recent years, with the development of synthetic biology, A variety of microbial cell factories have been constructed to synthesize plant natural products, which present advantages over their chemical synthesis. In this article, the classification, functional activities, structural characteristics of typical pentacyclic triterpenoids and the progress of their synthesis through microbial cell factories are reviewed based on their plant sources and natural synthesis pathway. The unknown enzymes catalyzing synthesis of the key sites of sapogenins in the pentacyclic triterpenoid saponins synthesis pathway are analyzed. Combined with the reported in vivo and in vitro studies, P450 enzymes and glycosyltransferases that are responsible for the formation of sapogenins and saponins, respectively, in some unidentified pathways are predicted. Based on current research, main bottlenecks for pentacyclic triterpenoid saponins synthesis in microbial cell factories are highlighted, and the situation of industrial production and challenges are discussed, which provide theoretical support and new ideas for constructing microbial cell factories to synthesize pentacyclic triterpenoid saponins more efficiently.

Key words: synthetic biology, pentacyclic triterpenoid saponins, microbial cell factory, glycosylation, synthetic pathway

摘要：

五环三萜皂苷类化合物具有丰富的药理、生理活性，广泛应用于医药、功能食品、保健品、化妆品等领域。目前五环三萜皂苷类化合物的主要获取方式是植物提取，随着合成生物学的发展，利用微生物细胞工厂合成植物天然产物逐渐成为研究热点，它具有生产周期短、工艺简单、环境友好、条件温和等优势，是未来的发展方向。本文结合五环三萜皂苷类化合物的来源及其天然合成途径，综述了典型五环三萜皂苷类化合物的分类、功能活性、结构特点及目前利用微生物细胞工厂合成五环三萜皂苷类化合物的研究现状；分析了部分五环三萜皂苷类化合物合成途径当中的未解析的关键修饰位点及关键酶，并结合已报道的体内、体外研究，对部分未知途径当中催化母核形成苷元的P450酶以及对苷元进行糖基化修饰的糖基转移酶进行了合理预测；结合当前研究现状分析、总结、归纳了利用微生物细胞工厂合成五环三萜皂苷类化合物存在的主要瓶颈，讨论了现阶段工业化生产现状及利用生物合成进行工业化生产所面临的挑战，为高效合成五环三萜皂苷类天然产物的微生物细胞工厂构建提供了理论支持和新思路。

关键词: 合成生物学, 五环三萜皂苷, 微生物细胞工厂, 糖基化修饰, 合成途径

CLC Number:

REN Shichao, SUN Qiuyan, FENG Xudong, LI Chun. Biosynthesis of pentacyclic triterpenoid saponins in microbial cell factories[J]. Synthetic Biology Journal, 2022, 3(1): 168-183.

任师超, 孙秋艳, 冯旭东, 李春. 微生物细胞工厂合成五环三萜皂苷类化合物[J]. 合成生物学, 2022, 3(1): 168-183.

Figures/Tables 7

References 70

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化合物	宿主	产量	主要策略	文献
金盏花苷E	体外合成		转录组分析、大肠杆菌重组表达	[17]
金盏花苷E	酿酒酵母	未报道	转录组分析、共表达	[27]
3-O-葡萄糖-齐墩果酸	体外合成		cDNA表达文库筛选	[16]
28-O-葡萄糖-齐墩果酸	体外合成		cDNA表达文库筛选	[16]
3-O-葡萄糖-甘草次酸	酿酒酵母	26.31 mg/L	基因组分析、MVA强化	[23]
30-O-葡萄糖-甘草次酸	体外合成		基因组分析、大肠杆菌重组表达	[23]
3,30-O-双葡萄糖-甘草次酸	体外合成		基因组分析、大肠杆菌重组表达	[23]
GAMG	体外合成		转录组分析、大肠杆菌重组表达	[24]
GAMG	酿酒酵母	未报道	转录组分析、共表达	[27]
GL	酿酒酵母	791 μg/L	转录组分析、共表达	[27]
GL	体外合成		转录组分析、昆虫细胞重组表达	[25]
3-O-葡萄糖醛酸-苜蓿酸	体外合成		转录组分析、植物瞬时表达	[26]
大豆皂苷Ⅲ	体外合成		大肠杆菌重组表达、无细胞体系催化	[36]
大豆皂苷Ⅰ	体外合成		大肠杆菌重组表达、无细胞体系催化	[36]
熊果酸28-O-β-D-吡喃葡萄糖苷	酿酒酵母	痕量	转录组分析、大肠杆菌重组表达、酿酒酵母异源表达	[51]

化合物	宿主	产量	主要策略	文献
金盏花苷E	体外合成		转录组分析、大肠杆菌重组表达	[17]
金盏花苷E	酿酒酵母	未报道	转录组分析、共表达	[27]
3-O-葡萄糖-齐墩果酸	体外合成		cDNA表达文库筛选	[16]
28-O-葡萄糖-齐墩果酸	体外合成		cDNA表达文库筛选	[16]
3-O-葡萄糖-甘草次酸	酿酒酵母	26.31 mg/L	基因组分析、MVA强化	[23]
30-O-葡萄糖-甘草次酸	体外合成		基因组分析、大肠杆菌重组表达	[23]
3,30-O-双葡萄糖-甘草次酸	体外合成		基因组分析、大肠杆菌重组表达	[23]
GAMG	体外合成		转录组分析、大肠杆菌重组表达	[24]
GAMG	酿酒酵母	未报道	转录组分析、共表达	[27]
GL	酿酒酵母	791 μg/L	转录组分析、共表达	[27]
GL	体外合成		转录组分析、昆虫细胞重组表达	[25]
3-O-葡萄糖醛酸-苜蓿酸	体外合成		转录组分析、植物瞬时表达	[26]
大豆皂苷Ⅲ	体外合成		大肠杆菌重组表达、无细胞体系催化	[36]
大豆皂苷Ⅰ	体外合成		大肠杆菌重组表达、无细胞体系催化	[36]
熊果酸28-O-β-D-吡喃葡萄糖苷	酿酒酵母	痕量	转录组分析、大肠杆菌重组表达、酿酒酵母异源表达	[51]

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