合成生物技术制备脂肪族二元胺的研究进展

doi:10.12211/2096-8280.2020-054

摘要/Abstract

摘要：

合成生物学作为发展迅速的一门交叉学科，为构建高效的微生物细胞工厂、促进生物基产品的产业化制备提供了强有力的工具。二元胺作为一种重要的聚合单体，广泛应用于聚酯、聚氨酯、聚酰胺等高分子材料的合成中。本文针对C₃～C₅脂肪族二元胺（1,5-戊二胺、1,3-丙二胺、1,4-丁二胺）的生物合成，从途径设计与构建、关键结构元件的设计和改造、调控元件的挖掘与优化、辅因子合成和转运调控等模块的优化和系统集成等方面，综述了利用合成生物学策略改造大肠杆菌和谷氨酸棒状杆菌合成二元胺的现状，并从非粮生物质的利用和生物合成过程中CO₂的再循环利用两个方面阐述了提高二元胺合成过程中原子经济性的研究概况，展望了如何利用合成生物技术进一步优化二元胺合成细胞的性能，以促进生物基二元胺的产业化生产。

关键词: 合成生物学, 脂肪族二元胺, 材料单体, 细胞工厂, 非粮生物质, 过程经济性

Abstract:

As a rapidly developing interdiscipline, synthetic biology has provided powerful tools for the development of the efficient microbial cell factories to promote the industrial preparation of bio-based products. As important monomers, diamines have been widely used in the synthesis of polymeric materials such as polyester, polyurethane, polyamide, etc. The aliphatic diamines with 3—5 carbon atoms, including 1,3-propanediamine, 1,4-butanediamine, and 1,5-pentanediamine are considered to be promising alternatives to traditional fossil-fuel-based diamines. In this review, the current status of the art of the biosynthesis of the aliphatic diamines with 3—5 carbon atoms by engineered Escherichia coli or Corynebacterium glutamicum are discussed. Several synthetic biology strategies, such as the design and construction of biosynthetic pathways, the design and reconstruction of key structural elements, the mining and optimization of regulatory elements, the optimization of cofactor regulation modules or mass transport modules, and their system integration were focused on due to their application in the improvement of cell production capacity. Furthermore, the utilization of non-food biomass and the recycling of CO₂generated during the diamine production process to improve the atom economy of diamine synthesis are also reviewed. Finally, the optimization of diamine producers by using synthetic biotechnology to promote the industrial production of bio-based diamines is prospected.

Key words: synthetic biology, aliphatic diamines, material monomer, cell factories, non-food feedstock, process economy

中图分类号:

王昕, 王静, 陈可泉, 欧阳平凯. 合成生物技术制备脂肪族二元胺的研究进展[J]. 合成生物学, 2020, 1(1): 71-83.

WANG Xin, WANG Jing, CHEN Kequan, OUYANG Pingkai. Research progress in bioproduction of aliphatic diamines by synthetic biotechnology[J]. Synthetic Biology Journal, 2020, 1(1): 71-83.

图/表 5

图1 C3～C5脂肪族二元胺的生物合成途径[8,9,10,11,12,13]（实线箭头为一步代谢途径,虚线箭头为一步以上代谢途径。绿色为1,3-丙二胺合成途径，紫色为1,4-丁二胺合成途径，蓝色为1,5-戊二胺合成途径）

Fig. 1 The biosynthetic pathway of the aliphatic diamines with 3～5 carbon atoms (The solid arrows indicate one-step metabolic pathways, and the dotted arrows indicate multiple-step metabolic pathways. The green square represents the synthetic pathway of 1,3-propanediamine, the purple square represents the synthetic pathway of 1,4-butanediamine synthesis route, and the blue square represents the synthetic pathway of 1,5-pentanediamine)

图2 二元胺合成细胞的优化策略 [14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48]

Fig. 2 The engineering strategy for the optimization of diamine synthetic cells[14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48]

表1 不同宿主合成C3～C5脂肪族二元胺最高水平对比

Tab. 1 The highest production level of aliphatic diamines with 3～5 carbon atoms by different hosts

产品	宿主	改造策略	产量 /g·L^-1	产率/g·g^-1	参考文献
1,5- 戊二胺	大肠杆菌	过表达赖氨酸脱羧酶元件cadA 过表达dapA基因增强前体赖氨酸合成敲除1,5-戊二胺利用途径基因speE、speG、ygjG和 puuA 抑制调控元件murE的表达	12.6	—	[38]
1,5- 戊二胺	谷氨酸棒状杆菌	高强度表达E.coli 来源的赖氨酸脱羧酶元件ldcC 敲除赖氨酸分泌蛋白lysE	103.78	0.303	[36]
1,4- 丁二胺	大肠杆菌	敲除产物降解途径基因 speE、speG、puuA 敲除前体鸟氨酸降解途径基因argI 敲除全局转录因子 rpoS 过表达合成途径基因speC、argE、argC、argB、argH、speF和argD 过表达1,4-丁二胺分泌蛋白potE 下调argF和glnA的表达水平	42.3	0.26	[39]
1,4- 丁二胺	谷氨酸棒状杆菌	异源表达 E. coli 来源的鸟氨酸脱羧酶speC 敲除前体鸟氨酸利用途径基因argF和argR 精确调控argF的表达水平	19	0.16	[37]
1,3- 丙二胺	大肠杆菌	表达不动杆菌属来源的dat和ddc组装1,3-丙二胺合成途径过表达thrA和lysC突变体增强天冬氨酸半醛合成过表达ppc和aspC增强前体天冬氨酸的合成敲除pfkA增强胞内NADPH的供给	13.06	0.1	[13]

图3 以非粮生物质为原料的生物基二元胺合成[50,51,52,53,54,55,56,57,58]

Fig. 3 The biosynthesis of aliphatic diamines with non-food feedstock

图4 生物基二元胺合成过程CO2的原位固定

Fig. 4 The in situ fixation of CO2 during the synthesis of bio-based diamines

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