Synthetic biology drives the sustainable production of terpenoid fragrances and flavors

doi:10.12211/2096-8280.2024-057

Abstract

Abstract:

The demand for personal care products has been steadily increasing. Consumers are now seeking products that offer enhanced functionality, natural ingredients, and superior sensory experiences. Fragrances and flavors are the key components in personal care formulations. Terpenes and their derivatives dominate natural fragrances due to their diverse structures and scents, widespread availability in plants and animals, structural stability, and high safety profile, making them highly marketable. The terpene fragrance market is projected to grow at a compound annual growth rate of 6.4%, reaching $1.01 billion by 2028, indicating a high market value and promising future. Currently, the acquisition of natural terpene fragrances is constrained by long plant growth cycles, low terpene content, and high extraction costs. Thus, there is an urgent need for developing new technology, such as synthetic biology, to achieve large-scale production of diverse fragrance compounds at an environment-friendly manner. This review explores the application and development of synthetic biology in the sustainable production of terpene fragrances. It highlights how data-driven synthetic biology and biotechnological innovations empower terpene fragrance production. This review compares classical and alternative terpenoid biosynthesis pathways, elucidating their differences and advantages. This analysis may offer comprehensive insights for chassis design toward terpenoid efficient biosynthesis. Additionally, this review explores recent advances in terpene synthase discovery and engineering as well as cell factory construction. This review comprehensively summarizes the common challenges encountered in the construction of three major types of terpene fragrance cell factories: monoterpenes, sesquiterpenes, and nor-isoprenoids. It also discusses the corresponding metabolic engineering strategies employed to address these issues, including enzyme optimization, pathway reconstruction, and cellular detoxification. Additionally, the review summarizes the current patent landscape and industrial competition, and it offers insights into future challenges and opportunities, including the hurdles of biosynthesis technology, the discovery and design of new products, as well as market regulation and safety concerns.

Key words: synthetic biology, terpenoid, fragrances and flavors, cell factory, intellectual property

摘要：

香精香料是个人护理产品中的重要成分，其中，萜类化合物及其衍生物在天然香料市场中有着重要的地位。近年来，合成生物学的蓬勃发展为解决萜类香料产能瓶颈及开发更多元化的新型香料化合物带来了新机遇。本综述探讨了合成生物学在萜类香料可持续生产中的应用和发展，介绍了数据驱动的合成生物学和生物技术创新如何赋能萜类香料生产，讨论比较了萜类合成的经典合成途径和替代合成途径，并探讨了萜类合酶挖掘与改造进展。在此基础上，着重介绍了单萜类、倍半萜类和降异戊二烯类香料的细胞工厂合成现状，包括元件改造、途径优化和萜类解毒等关键技术策略。最后，对当前专利布局和产业化竞争格局进行了总结分析，并对未来发展的挑战和机遇进行了展望，包括生物合成技术的挑战、新产物的发掘与设计，以及市场监管与安全性问题。

关键词: 合成生物学, 萜类化合物, 香精香料, 细胞工厂, 知识产权

CLC Number:

Q816

Mengyao ZHANG, Peng CAI, Yongjin ZHOU. Synthetic biology drives the sustainable production of terpenoid fragrances and flavors[J]. Synthetic Biology Journal, DOI: 10.12211/2096-8280.2024-057.

张梦瑶, 蔡鹏, 周雍进. 合成生物学助力萜类香精香料可持续生产[J]. 合成生物学, DOI: 10.12211/2096-8280.2024-057.

Figures/Tables 9

Fig. 1 The data- and biotechnology-driven 'Design-Build-Test-Learn' cycle in synthetic biology

Fig. 2 Classical terpene biosynthetic pathways and common alternative synthetic routes. Abbreviations: G3P, D-Glyceraldehyde 3-phosphate; PYR, pyruvate; DXP, 1-Deoxy-D-xylulose 5-phosphate; MEP, 2-C-Methyl-D-erythritol 4-phosphate; CDP-ME, 4-Diphophocytidyl-2-C-methyl-D-erythrito; CDP-MEP, 4-Diphophocytidyl-2-C-methyl-D-erythritol 2phosphate; MEcPP, 2-C-Methyl-D-erythritol 2,4-cyclodiphosphate; HMBPP, 4-Hydroxy-3-methyl-butenyl diphosphate; DMAPP, Dimethylallyl diphosphate; GPP, Geranyl diphosphate; FPP, Farnesyl diphosphate; GGPP, Geranylgeranyl diphosphate; MG-CoA, 3-Methylglutaconyl-CoA; MB-CoA, 3-Methyl-2-butenoyl-CoA; MB, 3-Methyl-2-butenal; DMAP, Dimethylallyl phosphate; M3P, Mevalonate 3-phosphate; M3P5P, Mevalonate 3,5-biphosphate; IP, Isopentenyl phosphate; AcCoA, Acetyl-CoA; AcAcCoA, Acetoacetyl-CoA; HMGCoA, 3-Hydroxy-3-methylglutaryl-CoA; MVA, Mevalonate; M5P, Mevalonate 5-phosphate; MVAPP, Mevalonate diphosphate; IPP, Isopentenyl diphosphate; NPP, Neryl diphosphate; Z,Z-FPP, Z,Z-Farnesyl diphosphate; NNPP, Nerylneryl diphosphate; DXS, DXP synthase; DXR, DXP reductoisomerase; MCT, MEP cytidylyltransferase; CMK, CDP-ME kinase; MDS, ME-CPP synthase; HDS, HMB-PP synthase; HDR, HMB-PP reductase; ERG10, ACCT acetyl-CoA C-acetyl transferase; ERG13, HMGS HMG-CoA synthase; HMGR, HMG-CoA reductase; ERG12, MK MVA kinase; ERG8, PMVK phosphomevalonate kinase; ERG19, MVD diphosphomevalonate decarboxylase; LiuC, enoyl-CoA hydratase; AibAB, glutaconyl-CoA decarboxylase; cbjALD, acyl-CoA reductase; YahK, alcohol dehydrogenase; ThiM, hydroxyethylthiazole kinase; IPK, isopentenyl phosphate kinase; IDI, isopentenyl-diphosphate isomerase; GPPS, geranyl pyrophosphate synthase; FPPS, farnesyl pyrophosphate synthase; GGPPS, geranylgeranyl diphosphate synthase; NPPS, nerol pyrophosphate synthase; zFPPS, Z,Z-Farnesyl diphosphate synthase; NNPPS, nerylneryl diphosphate synthase; M3K, mevalonate 3-kinase; M3P5K, mevalonate 3-phosphate 5-kinase; BMD, mevalonate biphosphate decarboxylase; IPK, isopentenyl phosphate kinase; PMD, mevalonate 5-phosphate decarboxylase.

Fig. 3 Mining and engineering terpene synthases.

Tab. 1 Summary of classic cell factories for terpene fragrances

结构	化合物	气味特征	底盘菌株	培养条件	产量（g/L）	参考文献
链状单萜	香叶醇 Geraniol	温和、甜香、花果香气	大肠杆菌	摇瓶发酵	2.1	[73]
	香叶醇 Geraniol	温和、甜香、花果香气	巴斯德毕赤酵母	24孔板发酵	1.2	[74]
	月桂烯 Myrcene	甜香脂香气	大肠杆菌	摇瓶发酵	1.2	[75]
	香茅醇 Citronellol	甜润玫瑰花香	酿酒酵母	5 L生物反应器	8.3	[76]
	芳樟醇 Linalool	铃兰清香	菠萝潘托氏菌	5 mL试管	S型5.6 R型3.7	[77]

Fig. 4 Common engineering strategies for monoterpene fragrance cell factories.

Fig. 5 Common strategies to enhance FPP availability for sesquiterpene synthases.

Fig. 6 Schematic diagram of biosynthetic pathway of α, β-ionone.

Fig. 7 Global distribution of literature and patents related to terpene fragrance biosynthesis in the last decade. (the data is from Google Patents and Google Scholar using "terpene fragrances" and "synthetic biology" as the keywords, July 17th, 2024).

Fig. 8 The current landscape of synthetic biology companies and traditional fragrance industries

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