Synthetic Biology Journal ›› 2023, Vol. 4 ›› Issue (5): 932-946.DOI: 10.12211/2096-8280.2023-035
• Invited Review • Previous Articles Next Articles
Zhehui HU1,2, Juan XU2, Guangkai BIAN1
Received:
2023-05-06
Revised:
2023-07-24
Online:
2023-11-15
Published:
2023-10-31
Contact:
Guangkai BIAN
胡哲辉1,2, 徐娟2, 卞光凯1
通讯作者:
卞光凯
作者简介:
基金资助:
CLC Number:
Zhehui HU, Juan XU, Guangkai BIAN. Application of automated high-throughput technology in natural product biosynthesis[J]. Synthetic Biology Journal, 2023, 4(5): 932-946.
胡哲辉, 徐娟, 卞光凯. 自动化高通量技术在天然产物生物合成中的应用[J]. 合成生物学, 2023, 4(5): 932-946.
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URL: https://synbioj.cip.com.cn/EN/10.12211/2096-8280.2023-035
应用领域 | 传统方法 | 自动化高通量方法 | ||
---|---|---|---|---|
方式 | 特点 | 方式 | 特点 | |
天然产物的挖掘 | 直接分离 同源激活 异源表达 | 生物样本单一 局限于单个基因的异源表达 产物丰度低、筛选通量低 | 打造特定微生物高产底盘 基于海量数据库批量挖掘 高通量工作站自动化验证 | 不受限于特定的生物样本 充分释放酶的产物合成潜力 规模化、自动化 |
天然产物高效合成 | 突变体文库平板筛选 关键酶的定点诱变 人工改造代谢通路 | 突变体间微小差异分辨率低 试错过程需要消耗大量劳动力 无法完全释放酶的催化潜力 | 基于auto-HTP对突变体文库高效筛选 自动化改造底盘细胞 限速酶定向进化 微生物适应性进化 | 自动化实现海量试错 多维度改造底盘细胞 缩短菌株改造研发周期 |
天然产物的快速检测 | 气相色谱、质谱 液相色谱、质谱 核磁共振 基于紫外/可见光谱检测 | 耗时长、通量低 对目标产物的产量和纯度要求高 | 基于荧光光谱检测 基于生物传感器的荧光检测 基于先进光谱技术的检测 | 便捷、高效、省时 对目标产物的动态、实时、自动化检测 |
Table 1 A comparison of automated high-throughput and traditional methods in natural product biosynthesis
应用领域 | 传统方法 | 自动化高通量方法 | ||
---|---|---|---|---|
方式 | 特点 | 方式 | 特点 | |
天然产物的挖掘 | 直接分离 同源激活 异源表达 | 生物样本单一 局限于单个基因的异源表达 产物丰度低、筛选通量低 | 打造特定微生物高产底盘 基于海量数据库批量挖掘 高通量工作站自动化验证 | 不受限于特定的生物样本 充分释放酶的产物合成潜力 规模化、自动化 |
天然产物高效合成 | 突变体文库平板筛选 关键酶的定点诱变 人工改造代谢通路 | 突变体间微小差异分辨率低 试错过程需要消耗大量劳动力 无法完全释放酶的催化潜力 | 基于auto-HTP对突变体文库高效筛选 自动化改造底盘细胞 限速酶定向进化 微生物适应性进化 | 自动化实现海量试错 多维度改造底盘细胞 缩短菌株改造研发周期 |
天然产物的快速检测 | 气相色谱、质谱 液相色谱、质谱 核磁共振 基于紫外/可见光谱检测 | 耗时长、通量低 对目标产物的产量和纯度要求高 | 基于荧光光谱检测 基于生物传感器的荧光检测 基于先进光谱技术的检测 | 便捷、高效、省时 对目标产物的动态、实时、自动化检测 |
Fig. 1 A comparison of different screening methods[41-42](a) Traditional screening method based on agar plates, with a screening throughput of 103~104; (b) Microplate screening method based on automation equipment, with a screening throughput of 104~105; (c) Fluorescence activated cell sorting, with a screening throughput of 108~109; (d) Droplet-based microfluidic sorting, with a screening throughput of 108~109
Fig. 2 Automated high-throughput workstation(a) Schematic diagram of the automated high-throughput workstation; (b) A representative workflow of automated high-throughput process, with yeast engineering shown as an example
Fig. 3 The application of biocasting in efficient mining of new functional terpenoid genes (gene clusters) and the biosynthesis of mangicol J, a new anti-inflammatory molecule[51, 56](a) The batch mining of 34 new chimeric terpenoid synthases (PTTC); (b) The reconstruction of 39 terpenoid gene clusters into 208 mutants; (c) High-throughput screening of 185 terpenoids leading to the isolation of compound 70 (mangicol J) with high anti-inflammatory activity; (d) The elucidation of the biosynthetic pathway of mangicol J; (e) The construction of a general Aspergillus oryzae chassis for efficient biosynthesis of mangicol J
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