From CO2 to value-added products—carbon neutral microalgal green biomanufacturing

doi:10.12211/2096-8280.2022-023

Abstract

Abstract:

Currently, our world is facing the dual pressure of carbon emission reduction and resource shortage. China has also put forward a goal of reaching CO₂ emission peak by 2030 and achieving carbon neutrality by 2060. At present, the production and manufacture of fuels and bulk chemical products mainly rely on petrochemical refining, which is facing the challenges of high risk of production safety, great pressure of environmental protection, and contradiction between supply and demand of oil and gas resources. In this context, the use of microalgae for direct CO₂ fixation is expected to establish large-scale biomanufacturing with CO₂ as raw material and sunlight as energy source, this is a new manufacturing mode that breaks away from the route of petrochemical industry, and has the typical characteristics of low carbon, recyclable, green, and clean. This emerging green industry is of strategic significance for solving the current issues of food security and energy shortages, through sustainable production of food, energy, chemicals, and pharmaceuticals. In addition, microalgae possess great potential in environment protection, thanks to their strong stress resistance, effective remediation of eutrophic elements such as nitrogen and phosphorus from wastewater, and simultaneous removal of SO _x and NO _x during CO₂ utilization in flue gas. Therefore, compared with heterotrophic chassis cells, microalgae-based synthetic biology and bio-manufacturing also play a role in carbon sequestration and emission reduction, and microalgae have then attracted much attention in recent years as “green cell factories”. From the perspective of light-driven autotrophy, we summarize the latest progress of microalgae as a cell factory, introduce chassis transformation strategies, and then look into the future development of this technology. In particular, improved genetic manipulation and larger cultivation scales are critical for microalgae to serve as high efficiency chassis for synthetic biology, whereas promising directions include establishment of standardized systems for algal genome editing, deep understanding of metabolic flux and control for robust biosynthesis, as well as improvement of biomass productivity and photosynthesis efficiency. All in all, this review provides a useful reference to establish controllable and replicable processes for microalgae green biomanufacturing.

Key words: microalgae, carbon dioxide, carbon platform compound, bio-energy, high-value compounds, synthetic biology

摘要：

微藻可以利用太阳能固定CO₂并转化为有机物，其作为合成生物物质的细胞工厂具有众多生物学和工程学优点。当前，全球正面临着碳减排和资源短缺的双重压力，通过微藻固碳合成化合物技术的攻关和突破，实现直接利用微藻固定CO₂，有望建立以CO₂为原料、以太阳能为能源，规模化生产大宗食物、能源、化学品和医药保健品的未来新兴绿色生物制造产业，对于解决当前面临的粮食安全、环境污染和能源紧缺等问题具有战略意义。本文从光驱自养的角度，首先总结了微藻作为细胞工厂生产平台化合物、生物能源和高附加值化合物的途径、底盘改造策略等最新进展，进而对该技术的未来发展方向进行展望。最后，提出了微藻作为合成生物学高效底盘细胞，其广泛应用还应该从建立标准化的藻类基因与基因组编辑技术体系、深刻理解合成物质在藻细胞中的代谢流和控制机制以及提高生物量产率和光合作用效率等几个环节进行攻关，以加强微藻绿色生物制造产业的可控性和可复制性。

关键词: 微藻, CO₂, 平台化合物, 生物能源, 高附加值化合物, 合成生物学

CLC Number:

Q816

Zhongliang SUN, Hui CHEN, Qiang WANG. From CO₂ to value-added products—carbon neutral microalgal green biomanufacturing[J]. Synthetic Biology Journal, 2022, 3(5): 953-965.

孙中亮, 陈辉, 王强. 从CO₂到有机物——碳中和的微藻绿色生物制造[J]. 合成生物学, 2022, 3(5): 953-965.

Figures/Tables 6

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表达产物	合成聚合物	表达株系	最终产率/[mg/(L·d)]	参考文献
1，3-丙二醇	聚酯、聚醚、聚氨酯、PTT	Synechococcus sp. PCC 7942	61	[25]
		Synechococcus sp. PCC 7942	8	[33]
		Synechococcus sp. PCC 7942	20.6	[34]
		Synechococcus sp. PCC 7120	2.3	[26]
1，2-丙二醇	聚丙二醇	Synechococcus sp. PCC 7942	15	[27]
		Synechocystis sp. PCC 6803	100	[28]
异戊二烯	橡胶	Synechocystis sp. PCC 6803	60	[30]
		Synechocystis sp. PCC 6803	1.7	[35]
乙烯	聚乙烯、聚苯乙烯、PVC、聚酯	Synechocystis sp. PCC 6803	2104	[36]
		Synechocystis sp. PCC 6803	19.2	[37]
3-羟基丙酸	聚3-羟基丙酸	Synechocystis sp. PCC 6803	139.5	[38]
		Synechococcus sp. PCC 7942	329.5	[39]
2，3-丁二醇	树脂	Synechococcus sp. PCC 7942	54.36	[40]
		Synechococcus sp. PCC 7942	300	[41]
		Synechococcus sp. PCC 7942	110	[40]
		Synechococcus sp. PCC 7942	100	[42]

表达产物	合成聚合物	表达株系	最终产率/[mg/(L·d)]	参考文献
1，3-丙二醇	聚酯、聚醚、聚氨酯、PTT	Synechococcus sp. PCC 7942	61	[25]
		Synechococcus sp. PCC 7942	8	[33]
		Synechococcus sp. PCC 7942	20.6	[34]
		Synechococcus sp. PCC 7120	2.3	[26]
1，2-丙二醇	聚丙二醇	Synechococcus sp. PCC 7942	15	[27]
		Synechocystis sp. PCC 6803	100	[28]
异戊二烯	橡胶	Synechocystis sp. PCC 6803	60	[30]
		Synechocystis sp. PCC 6803	1.7	[35]
乙烯	聚乙烯、聚苯乙烯、PVC、聚酯	Synechocystis sp. PCC 6803	2104	[36]
		Synechocystis sp. PCC 6803	19.2	[37]
3-羟基丙酸	聚3-羟基丙酸	Synechocystis sp. PCC 6803	139.5	[38]
		Synechococcus sp. PCC 7942	329.5	[39]
2，3-丁二醇	树脂	Synechococcus sp. PCC 7942	54.36	[40]
		Synechococcus sp. PCC 7942	300	[41]
		Synechococcus sp. PCC 7942	110	[40]
		Synechococcus sp. PCC 7942	100	[42]

藻株	操作方法	有益结果	油脂含量或产率	参考文献
Phaeodactylum tricornutum	过表达内源性苹果酸脱氢酶	油脂含量增加250%	57.8%	[50]
Chlamydomonas reinhardtii	Lobosphaera incise中的GPAT基因（LiGPAT）在莱茵衣藻中过表达	TAG含量比野生型增加50%	50.0%	[51]
Thalassiosira pseudonana	转化反义结构体，阻止脂质分解代谢	油脂含量比野生型增加230%	18.8%	[52]
Phaeodactylum tricornutum	过表达Ⅱ型甘油二酯酰基转移酶（DGAT2D）基因	TAG含量增加35%	37.2%	[53]
Chlorella spp	拟南芥激酶的上调和过表达	油脂产量增加110.4%	27.5%	[54]
Nannochloropsis	过表达新型bZIP1转录因子NobZIP1N	油脂含量增加而不影响微藻生长	40%	[55]
Chlamydomonas reinhardtii	使用CRISPR-Cas9敲除磷脂酶A2基因	油脂含量可达64.25%	80.92 mg/(L·d)	[56]
Chlamydomonas reinhardtii	使用CRISPR-Cas9 RNP方法产生葡萄糖焦磷酸化酶基因（AGP）突变的突变体	油脂含量比野生型增加274%	57.67%	[56]
Nannochloropsis	通过插入突变产生突变体Mut68	脂肪酸甲酯含量和产量分别比野生型增加34%和75%	78.3 mg/(L·d)	[57]
Planktochlorella nurekis	利用细胞松弛素B和秋水仙碱调控DNA水平	油脂含量比野生型增加10%～60%	12%～26%	[58]

藻株	操作方法	有益结果	油脂含量或产率	参考文献
Phaeodactylum tricornutum	过表达内源性苹果酸脱氢酶	油脂含量增加250%	57.8%	[50]
Chlamydomonas reinhardtii	Lobosphaera incise中的GPAT基因（LiGPAT）在莱茵衣藻中过表达	TAG含量比野生型增加50%	50.0%	[51]
Thalassiosira pseudonana	转化反义结构体，阻止脂质分解代谢	油脂含量比野生型增加230%	18.8%	[52]
Phaeodactylum tricornutum	过表达Ⅱ型甘油二酯酰基转移酶（DGAT2D）基因	TAG含量增加35%	37.2%	[53]
Chlorella spp	拟南芥激酶的上调和过表达	油脂产量增加110.4%	27.5%	[54]
Nannochloropsis	过表达新型bZIP1转录因子NobZIP1N	油脂含量增加而不影响微藻生长	40%	[55]
Chlamydomonas reinhardtii	使用CRISPR-Cas9敲除磷脂酶A2基因	油脂含量可达64.25%	80.92 mg/(L·d)	[56]
Chlamydomonas reinhardtii	使用CRISPR-Cas9 RNP方法产生葡萄糖焦磷酸化酶基因（AGP）突变的突变体	油脂含量比野生型增加274%	57.67%	[56]
Nannochloropsis	通过插入突变产生突变体Mut68	脂肪酸甲酯含量和产量分别比野生型增加34%和75%	78.3 mg/(L·d)	[57]
Planktochlorella nurekis	利用细胞松弛素B和秋水仙碱调控DNA水平	油脂含量比野生型增加10%～60%	12%～26%	[58]

藻株	操作方法	有益结果	氢气产量或产率	参考文献
Chlamydomonas reinhardtii	设计光诱导系统，利用该系统设计蓝光诱导产氢转基因藻类	成功激活了人工miRNA，提高微藻的产氢能力	20 μL/(L·h)	[62]
Chlamydomonas reinhardtii	将大肠杆菌的丙酮酸氧化酶和过氧化氢酶基因整合到衣藻叶绿体基因组中	生物氢产量增加了2倍	1.04 μmol/(L·h)	[63]
Chlorella sp. DT.	敲除psbO基因	生物氢产量比野生型提高了9倍	350 mL/L	[64]
Chlamydomonas reinhardtii	截短捕光天线	氢气产量比野生型增加6倍	30 mL/L	[65]
Synechocystis sp. PCC 6803	添加抑制光合和呼吸作用电子传递链的抑制剂	氢气产量增加了30倍	1.25 μmol/(L·h)	[66]
Chlamydomonas reinhardtii	热诱导人工miRNA表达系统	氢气合成增加60%	90 μL/mg Chl	[67]

From CO₂ to value-added products—carbon neutral microalgal green biomanufacturing

从CO₂到有机物——碳中和的微藻绿色生物制造

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分类	藻株	合成产物	参考文献
原核藻	Synechocystis sp. PCC 6803	对香豆酸	[81]
	Synechocystis sp. PCC 6803	叶黄素	[82]
	Synechococcus elongatus UTEX 2973	柠檬烯	[83]
	Synechocystis sp. PCC 6803	乳酸	[84]
	Synechococcus sp. PCC 7002	虾青素	[85]
	Anabaena sp. PCC7120	法尼烯	[86]
	Synechococcus sp. PCC 7942	柠檬烯	[87]
真核藻	Chlamydomonas reinhardtii	类异戊二烯	[88]
	Chlamydomonas reinhardtii	木糖醇	[89]
	Chlamydomonas reinhardtii	倍半萜	[90]
	Phaeodactylum tricornutum	DHA	[91]
	Bacillariophyta	没药烯	[92]
	Nannochloropsis	EPA	[93]
	Phaeodactylum tricornutum	岩藻黄质	[94]

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