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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2024, Vol. 44 Issue (1): 72-87    DOI: 10.13523/j.cb.2312106
    
Challenges in the Biosynthesis of Natural Products and Microbial Manufacturing
Qiuyang LI1,2,Wentao SUN2,3,Lei QIN2,4,Bo LV1,4,**(),Chun LI1,2,4,**()
1 Key Laboratory of Medicinal Molecule Science and Pharmaceutical Engineering (Ministry of Industry and Information Technology), Institute of Biochemical Engineering, School of Chemistry and Chemical Engineering,Beijing Institute of Technology, Beijing 100081, China
2 Department of Chemical Engineering, Tsinghua University, Beijing 100084, China
3 State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica,Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China
4 Key Lab for Industrial Biocatalysis (Ministry of Education), Tsinghua University, Beijing 100084, China
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Abstract  

Natural products are a treasure trove given to mankind by nature and are intimately linked to human quality of life and health. Traditional methods of obtaining natural products face challenges such as low natural biocontent, complex and time-consuming extraction processes, and unsustainable resource utilization. The intricate structures of natural products also severely impede the commercialization of chemical total synthesis. Microbial cell factories, empowered by synthetic biology, have the potential to revolutionize the current approaches to natural product extraction, with the goal of efficient, eco-friendly, and controllable biomanufacturing. This review discusses the challenges posed by traditional methods of obtaining natural products and summarizes the characteristics of different types of cell factories along with examples of natural product synthesis. Considering the complexity of natural product metabolic pathways and issues such as unpredictable product structure and properties, low purity, and low yield due to the stability of chassis cells, the article analyzes the challenges in designing microbial cell factories from the perspective of catalytic component limitations, pathway assembly regulation, and chassis cell network remodeling. Strategies and countermeasures are discussed. Moreover, recommendations are proposed to address the current regulatory constraints in microbial manufacturing.

Key words： Natural products      Biomanufacturing      Microbial cell factories      Chassis cells      Biosafety     
Received: 11 January 2024      Published: 04 February 2024
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Qiuyang LI
Wentao SUN
Lei QIN
Bo LV
Chun LI
Cite this article:

Qiuyang LI, Wentao SUN, Lei QIN, Bo LV, Chun LI. Challenges in the Biosynthesis of Natural Products and Microbial Manufacturing. China Biotechnology, 2024, 44(1): 72-87.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2312106     OR     https://manu60.magtech.com.cn/biotech/Y2024/V44/I1/72

Fig.1 Development process of microbial cell factories for the production of natural products
类型 分类 名称 天然来源 功效 参考文献
萜烯类 单萜 柠檬烯 柑橘属果皮 抗炎、抗菌、镇静、促进消化、减肥 [11]
香叶醇 玫瑰 抗氧化、抗菌、保肝、神经保护 [12]
倍半萜 青蒿素 青蒿 抗疟疾 [13]
红没药烯 香柠檬 抗痒、消炎、抗癌 [11]
二萜 紫杉醇 红豆杉 抗肿瘤 [14]
香紫苏醇 抹香鲸/香紫苏 抗菌消炎、利胆、抗癌 [15]
三萜 甘草酸 甘草 免疫调节、抗肿瘤、抗病毒、保肝 [16]
原人参二醇 人参 镇静、促进细胞分化增殖、抗肿瘤、降血糖 [17]
齐墩果酸 丁香/女贞子、三七 抗HIV、抗菌、抗癌、抗溃疡、治疗骨质疏松 [18]
四萜 虾青素 雨生红球藻/红发夫酵母 抗氧化、抗炎、伤口愈合、保护心脏、保肝、抗糖尿病、神经保护、抗癌 [19]
番茄红素 番茄 抗氧化、保护心血管 [20]
黄酮类 黄酮 黄芩素 黄芩 保肝、镇静、抗过敏、抗炎 [21]
二氢黄酮 柚皮素 葡萄柚 抗氧化、抗雄激素、消炎、抗癌 [22]
黄酮醇 淫羊藿素 淫羊藿 抗肿瘤、神经保护、心血管保护、免疫调节、骨保护 [23]
生物碱 异喹啉类 吗啡 罂粟 治疗急性和慢性中度至重度疼痛 [24]
四氢异喹啉类 海鞘素 加勒比海鞘 抗肿瘤 [25]
单萜吲哚类 长春碱 长春花 抗肿瘤 [26]
酚酸 单羟基苯甲酸 水杨酸 柳树 缓解疼痛和发热、消炎、治疗痤疮 [27]
二羟基苯甲酸 原儿茶酸 乌蕨/冬青 抗氧化、抗肿瘤、神经保护 [28]
三羟基苯甲酸 没食子酸 五倍子/金缕梅 抗氧化、抗癌、抗菌、保护软骨、碳酸酐酶抑制剂、抗糖尿病、抗溃疡、组织蛋白酶D抑制剂 [29]
抗生素 聚酮类 四环素 链霉菌 治疗呼吸道、胃肠道和泌尿生殖道感染 [30]
大环内酯类 红霉素 糖多孢红霉菌 治疗呼吸道、皮肤和衣原体菌感染,治疗梅毒 [31]
Table 1 Beneficial effects of classic natural products
植物 提取部位 天然产物 提取方式
甘草 根部 甘草酸、光甘草定 水提取、醇提取
红豆杉 树皮 紫杉醇 乙醇、甲醇浸提
大黄 根部 大黄素 水提取、乙醇提取
丁香 花蕾 丁香酚 蒸馏提取、挥发油提取
当归 根部 川芎内酯、川芎酮 水提取、乙醇提取
葡萄柚 果皮 柚皮苷 挤压提取、冷榨提取
生姜 根部 姜辣素 水提取、乙醇提取
绿茶 叶 茶多酚、儿茶素 水提取
长春花 种子、块茎 长春碱 乙醇、甲醇浸提
Table 2 Traditional extraction methods for plant natural products
Fig.2 Types of microbial cell factories
类型 名称 天然来源 底盘细胞 产量 报道年份 参考文献
萜烯类 青蒿酸 黄花蒿 酿酒酵母 25 g/L 2013 [34]
α-檀香烯 檀香 毕赤酵母 21.5 g/L 2022 [35]
(+)-瓦伦烯 柑橘 酿酒酵母 16.6 g/L 2022 [36]
佛术烯 圣罗勒 酿酒酵母 34.6 g/L 2021 [37]
原人参二醇 人参 酿酒酵母 15.88 g/L 2022 [38]
α-香树脂醇 甘草 酿酒酵母 1 g/L 2020 [39]
11-氧-β-香树脂醇 甘草 酿酒酵母 108.1 mg/L 2018 [40]
甘草次酸 甘草 酿酒酵母 36 mg/L 2018 [40]
齐墩果酸 女贞 酿酒酵母 1 132.9 mg/L 2023 [41]
甘草酸 甘草 酿酒酵母 5.98 mg/L 2021 [42]
三七皂苷Rg1 三七 酿酒酵母 1.95 g/L 2021 [43]
番茄红素 番茄 解脂耶氏酵母 17.6 g/L 2022 [44]
虾青素 胡萝卜 解脂耶氏酵母 3.3 g/L 2022 [45]
黄酮类 野黄芩苷 黄芩 解脂耶氏酵母 703 mg/L 2023 [46]
柚皮素 葡萄柚 酿酒酵母 1 129.44 mg/L 2023 [47]
圣草酚 柠檬 酿酒酵母 3.28 g/L 2021 [48]
甘草素 甘草 解脂耶氏酵母 62.4 mg/L 2021 [49]
淫羊藿苷 淫羊藿 酿酒酵母 130 μg/L 2023 [50]
葛根素 野葛 酿酒酵母 72.8 mg/L 2021 [51]
大豆苷元 大豆 酿酒酵母 85.4 mg/L 2021 [51]
生物碱 莨菪碱 茄科植物 酿酒酵母 480 μg/L 2020 [52]
东莨菪碱 茄科植物 酿酒酵母 172 μg/L 2020 [52]
文朵灵 长春花 酿酒酵母 149.3 μg/L 2022 [53]
长春质碱 长春花 毕赤酵母 2.57 mg/L 2023 [10]
S-去甲乌药碱 南天竹 酿酒酵母-树干毕赤酵母 11.5 mg/L 2023 [54]
蒂巴因 罂粟 酿酒酵母 7.8 μg/L 2015 [55]
诺斯卡品 罂粟 酿酒酵母 2.2 mg/L 2018 [56]
Table 3 Natural products synthesized by yeast cell factories
类型 名称 天然来源 产量 报道年份 参考文献
萜烯 柠檬烯 柑橘属果皮 3.6 g/L 2020 [57]
香叶醇 玫瑰 2.12 g/L 2021 [58]
乙酸香叶酯 玫瑰 10.36 g/L 2022 [59]
紫穗槐二烯 黄花蒿 30 g/L 2019 [60]
贝壳杉烯 贝壳杉 623.6 mg/L 2020 [61]
紫杉烯 红豆杉 1 g/L 2010 [62]
5α-羟化紫杉烯醇 红豆杉 58 mg/L 2010 [62]
黄酮 芹菜素 芹菜 109.7 mg/L 2019 [63]
芫花素 芫花 41.0 mg/L 2015 [64]
黄芩素 黄芩 367.8 mg/L 2022 [65]
松属素 五针松 525.8 mg/L 2016 [66]
染料木素 槐角 35.1 mg/L 2022 [67]
Table 4 Natural products synthesized by E. coli cell factories
名称 结构类型 天然来源 产量 报道年份 参考文献
γ-氨基丁酸 氨基酸 广泛分布 45.6 g/L 2022 [68]
L-茶氨酸 氨基酸 茶叶 44.12 g/L 2023 [69]
血红素 铁卟啉 动物血液 309.18 mg/L 2021 [70]
β-胡萝卜素 四萜 胡萝卜 2.5 mg/g CDW 2023 [71]
虾青素 四萜 雨生红球藻/红发夫酵母 0.6 mg/g CDW 2023 [71]
Table 5 Natural Products of non-essential amino acids synthesized by C. glutamicum cell factories
Fig.3 Exploration and modification strategies for catalytic components
Fig.4 Constructing efficient cell factories through chassis cell remodeling and metabolic balance regulation
Fig.5 Workflow of retrobiosynthesis
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