Progress on bio-fixation and utilization of CO2 in acetogens driven by chemical energy

doi:10.12211/2096-8280.2023-045

Abstract

Abstract:

To promote the development of carbon utilization and biomanufacturing industry, one-carbon substrates are expected to be the next generation feedstocks and attracted much attention because of their abundance and availability for value-added products. Acetogens can naturally use syngas (CO, CO₂ and H₂) through the classical Wood-Ljungdahl pathway to support their growth and produce value-added chemicals such as acetic acid and ethanol. Compared to chemical catalysis, this biorefinery process has the advantages of low requirements for feedstocks, mild reaction conditions and high product selectivity. In some acetogens, both CO and H₂ can act as energy sources to provide the reducing equivalents required for growth and metabolism during gas fermentation. In particular, acetogens utilizing CO and H₂ have distinct patterns of energy metabolism. Compared to H₂, CO is a more thermodynamically favorable energy source for gas fermentation. In general, different acetogens during autotrophic growth with syngas as carbon source and energy source can produce different products. However, the low solubility of gases in liquid medium, as well as the poor mass transfer and diffusion, limits the energy supply during the fermentation, resulting in slow growth rate of microorganisms, low raw material utilization, and low target product yield. Therefore, improving syngas utilization efficiency and enlarging products spectrum of acetogens are both of great importance to meet the demands of industrialization. In recent years, with the development of synthetic biology and molecular genetic tools, modification of acetogen strains has been well designed. In this review, we briefly described the natural metabolic pathways such as ethanol and acetone metabolism pathways, and summarized the progress of acetogens in autotrophic fermentation using syngas as energy sources to produce value-added products such as ethanol, 2,3-butanediol, acetone and isoprepanol. For engineering the acetogens, we summarized the pathways to produce natural and non-natural chemicals. Finally, the energy capture and utilization of acetogens was prospected, with the aim to facilitate the future advancement of biorefineries.

Key words: acetogen, C₁-gas, chemoautotrophy, gene engineering, energy conservation

摘要：

利用微生物发酵一碳气体生产燃料和高值化学品，是实现碳资源回收利用和绿色生物制造的重要途径之一。产乙酸菌可利用CO或H₂为能量来源，通过伍德-永达尔（Wood-Ljungdahl）途径固定CO₂，维持自身代谢，并生产乙酸、乙醇等高附加值产品。相较于化学催化而言，该生物催化过程对原料气体比例要求不高、反应条件温和、产物选择性高。在气体发酵中，CO和H₂均可作为菌株生长和代谢的能量来源，二者具有不同的能量代谢模式。同时，不同的产乙酸菌代谢产物也不相同。发酵过程中气体溶解度低限制了能量供给，导致微生物生长速率慢、原料利用率低以及目标产物产量低等问题。提高气体利用效率和扩大产乙酸菌的产物谱，是面向产业化应用的必然要求。本文简述了产乙酸菌的伍德-永达尔途径、产乙酸途径和产乙醇途径等天然代谢途径，从分子改造的角度总结了产乙酸菌利用一碳气体发酵生产高附加值产品如乙醇、2，3-丁二醇、丙酮、异丙醇等的相关研究进展，并从提高能量供给和产物选择性的角度进行了总结。最后，本文以能量代谢为切入点对产乙酸菌转化一碳气体进行了展望，以期为未来的生物制造提供参考。

关键词: 产乙酸菌, 一碳气体, 化能自养, 基因工程, 能量守恒

CLC Number:

Zhiqiong WEN, Yuzhen LI, Jin′gang ZHANG, Feifei Wang, Xiaoqing MA, Fuli LI. Progress on bio-fixation and utilization of CO₂ in acetogens driven by chemical energy[J]. Synthetic Biology Journal, 2023, 4(6): 1178-1190.

文志琼, 李煜真, 张金刚, 王菲菲, 马小清, 李福利. 化能驱动的产乙酸菌转化利用CO₂研究进展[J]. 合成生物学, 2023, 4(6): 1178-1190.

Figures/Tables 5

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微生物	底物	产物	最适温度	最适pH	参考文献
Acetobacterium woodii	H₂/CO₂	乙酸	30	7.0	[9]
Clostridium aceticum	H₂/CO₂，CO	乙酸	30	8.3	[10-11]
Clostridium autoethanogenum	H₂/CO₂，CO	乙酸，乙醇，2，3-丁二醇	37	6.0	[12-13]
Clostridium carboxidivorans	H₂/CO₂，CO	乙酸，乙醇，丁酸，丁醇，己二醇	38	5.0～7.0	[14-15]
Clostridium ljungdahlii	H₂/CO₂，CO	乙酸，乙醇，2，3-丁二醇	37	6.0	[16-17]
Clostridium coskatii	H₂/CO₂，CO	乙酸，乙醇	37	6.0	[18]
Clostridium ragsdalei	H₂/CO₂，CO	乙酸，乙醇，2，3-丁二醇	37	6.3	[13,18]
Clostridium drakei	H₂/CO₂，CO	乙酸，乙醇	37	6.0	[19]
Sporomusa ovata	H₂/CO₂	乙酸	34	6.3	[20-21]
Thermoanaerobacter kivu	H₂/CO₂，CO	乙酸	66	6.4	[22-23]
Moorella thermoacetica	H₂/CO₂，CO	乙酸	55	7	[24-25]

微生物	底物	产物	最适温度	最适pH	参考文献
Acetobacterium woodii	H₂/CO₂	乙酸	30	7.0	[9]
Clostridium aceticum	H₂/CO₂，CO	乙酸	30	8.3	[10-11]
Clostridium autoethanogenum	H₂/CO₂，CO	乙酸，乙醇，2，3-丁二醇	37	6.0	[12-13]
Clostridium carboxidivorans	H₂/CO₂，CO	乙酸，乙醇，丁酸，丁醇，己二醇	38	5.0～7.0	[14-15]
Clostridium ljungdahlii	H₂/CO₂，CO	乙酸，乙醇，2，3-丁二醇	37	6.0	[16-17]
Clostridium coskatii	H₂/CO₂，CO	乙酸，乙醇	37	6.0	[18]
Clostridium ragsdalei	H₂/CO₂，CO	乙酸，乙醇，2，3-丁二醇	37	6.3	[13,18]
Clostridium drakei	H₂/CO₂，CO	乙酸，乙醇	37	6.0	[19]
Sporomusa ovata	H₂/CO₂	乙酸	34	6.3	[20-21]
Thermoanaerobacter kivu	H₂/CO₂，CO	乙酸	66	6.4	[22-23]
Moorella thermoacetica	H₂/CO₂，CO	乙酸	55	7	[24-25]

天然产物	微生物	原料	产量 /(g/L)	参考文献
Acetate	A. woodi	CO₂/H₂	51	[54]
	C. ljungdahlii	CO₂/H₂	35	[56]
Ethanol	C. ljungdahlii	CO/CO₂	32.8	[56]
	C. autoethanogenum	CO	2.46	[42]
	C. ljungdahlii	CO/CO₂/H₂	28.4	[57]
2,3-Butanediol	C. ljungdahlii	CO/CO₂	16.9	[56]
	C. autoethanogenum	CO/CO₂/H₂	0.2	[58]

天然产物	微生物	原料	产量 /(g/L)	参考文献
Acetate	A. woodi	CO₂/H₂	51	[54]
	C. ljungdahlii	CO₂/H₂	35	[56]
Ethanol	C. ljungdahlii	CO/CO₂	32.8	[56]
	C. autoethanogenum	CO	2.46	[42]
	C. ljungdahlii	CO/CO₂/H₂	28.4	[57]
2,3-Butanediol	C. ljungdahlii	CO/CO₂	16.9	[56]
	C. autoethanogenum	CO/CO₂/H₂	0.2	[58]

天然产物	微生物	策略	产量/产率	参考文献
丙酮	C. autoethanogenum	构建丙酮合成途径并强化ctfAB的表达	约3 g/(L·h)	[34]
异戊二烯	C. ljungdahlii	质粒共表达甲羟戊酸途径及异戊二烯合成酶基因	1.5 ng/mL	[62]
丁酸	C. ljungdahlii	敲除pta、adhE1和乙酰辅酶A转移酶基因，导入丁酸合成途径	1.32 g/L	[63]
异丙醇	C. ljungdahlii	外源引入异丙醇合成途径	13.4 g/L	[57]
3-羟基丁酸	C. ljungdahlii	外源引入3-羟基丁酸合成途径	3.0 g/L	[57]
聚羟基丁酸	C. autoethanogenum	共表达phaA、phaB、phaC	0.027 g/L	[67]

Progress on bio-fixation and utilization of CO₂ in acetogens driven by chemical energy

化能驱动的产乙酸菌转化利用CO₂研究进展

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