蓝藻细胞工厂的研究进展

doi:10.19586/j.2095-2341.2022.0203

生物技术进展 ›› 2023, Vol. 13 ›› Issue (2): 174-180.DOI: 10.19586/j.2095-2341.2022.0203

• 进展评述 • 上一篇

蓝藻细胞工厂的研究进展

赵俊魁(), 卢永忠()

青岛科技大学海洋科学与生物工程学院，山东青岛 266042

收稿日期:2022-11-03 接受日期:2023-01-06 出版日期:2023-03-25 发布日期:2023-04-07
通讯作者: 卢永忠
作者简介:赵俊魁 E-mail：happyzhaojk@163.com；
基金资助:
山东省重点研发计划项目(2019GHY112087)

Research Progress of Cyanobacteria Cell Factories

Junkui ZHAO(), Yongzhong LU()

College of Marine Science and Biological Engineering，Qingdao University of Science and Technology，Shandong Qingdao 266042，China

Received:2022-11-03 Accepted:2023-01-06 Online:2023-03-25 Published:2023-04-07
Contact: Yongzhong LU

摘要/Abstract

摘要：

蓝藻固碳能力强、易于进行遗传操作，是开发绿色生物技术产品的理想细胞工厂。但长期以来，许多研究成果仍处于实验室阶段，各种化学品生产的效率较低。结合蓝藻自身优势，充分利用现代生物技术手段，将有助于促进蓝藻细胞工厂的深入研究和发展。以蓝藻生物技术发展过程为主线，分别介绍了基因工程、代谢工程及合成生物学在蓝藻细胞工厂研究中取得的成果，并指出“药食同源”是蓝藻表达外源药物蛋白，实现成果转化的发展方向；开发先进的合成生物学工具，改造、设计和重构蓝藻底盘细胞代谢途径，是实现化学品高效生产的必由之路。期望为蓝藻生物技术相关问题的深入研究提供参考。

关键词: 蓝藻, 基因工程, 代谢过程, 化学品, 底盘细胞, 合成生物学

Abstract:

With strong carbon fixation capacity and amenability to genetic manipulations， cyanobacteria are considered to be ideal cell factories for developing green biotechnology products. However， many research achievements are still in the laboratory stage， and the production efficiency of various chemicals is low for a long time. Combining the advantages of cyanobacteria and making full use of modern biotechnology will help promote the in-depth research and development of cyanobacterial cell factories. Taking the development process of cyanobacterial biotechnology as the main line， this paper introduced the achievements of genetic engineering， metabolic engineering and synthetic biology in the research of cyanobacterial cell factory， and pointed out that "drug and food homology" was the direction of foreign protein expression by cyanobacteria for application. To develop advanced synthetic biological tools so as to transform， design and reconstruct the metabolic pathways of cyanobacteria chassis cells was the only way to achieve efficient production of chemicals. This paper was expected to provide a reference for the in-depth study of cyanobacterial biotechnology.

Key words: cyanobacteria, genetic engineering, metabolic process, chemical, chassis cells, synthetic biology

中图分类号:

赵俊魁, 卢永忠. 蓝藻细胞工厂的研究进展[J]. 生物技术进展, 2023, 13(2): 174-180.

Junkui ZHAO, Yongzhong LU. Research Progress of Cyanobacteria Cell Factories[J]. Current Biotechnology, 2023, 13(2): 174-180.

图/表 2

参考文献 55

1	JAISWAL D, SAHASRABUDDHE D, WANGIKAR P P. Cyanobacteria as cell factories: the roles of host and pathway engineering and translational research[J]. Curr. Opin. Biotechnol., 2022, 73: 314-322.
2	NG I S, KESKIN B B, TAN S I. A critical review of genome editing and synthetic biology applications in metabolic engineering of microalgae and cyanobacteria[J/OL]. Biotechnol. J., 2020, 15(8): e1900228[2022-12-20]. .
3	RUFFING A M, KALLAS T. Editorial: cyanobacteria: the green E . coli[J/OL]. Front. Bioeng. Biotechnol., 2016, 4: 7[2022-12-20]. .
4	席超,王春梅,施定基.蓝藻基因工程应用研究进展[J].中国生物工程杂志,2010,30(3):105-111.
5	徐旭东,孔任秋,胡玉祥.基因工程杀蚊幼蓝藻的研究[J].中国媒介生物学及控制杂志,1993,4(4):244-247.
6	任黎,邵强,施定基,等.人肝金属硫蛋白-IA基因在鱼腥藻中的克隆与表达[J].中国生物化学与分子生物学报,1998,14(4):15-21.
7	罗娜,宁叶,施定基,等.人尿激酶原基因在聚球藻7002中的克隆和表达[J].植物学报,2000,42(9):931-935.
8	施定基,叶欣,钟晖,等. TNFα基因在鱼腥藻7120中的表达及其产物的亲和层析纯化[J].植物学报,2001,43(1):46-50.
9	戴溦,施定基,张卉,等.人表皮生长因子(hEGF)基因在蓝藻中的表达[J].植物学报,2001,43(12):1260-1264.
10	刘仁海,高淑彬,章军,等.转hCu,Zn-SOD突变基因聚球藻抗氧化作用的研究[J].中国海洋药物,2007,26(6):10-12.
11	魏兰珍,谭玮,王全喜.启动子Pcpcβ提高鱼腥藻7120中hGM-CSF基因表达效率的研究[J].西北植物学报,2008,28(1):37-42.
12	王智,张泽峰,王晶晶,等.迟缓爱德华氏菌Eta1-L-Gapdh融合蛋白在蓝藻中的表达[J].微生物学杂志,2016,36(5):78-84.
13	郭媛媛,殷嵘,施定基,等.转vp28蓝藻口服剂对凡纳滨对虾抗白斑综合征病毒能力及免疫反应的影响[J].水产学报,2017, 41(9):1473-1485.
14	庄旻敏,贾晓会,施定基,等.转基因聚球藻7942中vp28基因表达效率及其光合特性分析[J].中国生物工程杂志,2018,38(4):30-37.
15	徐杨,谢京昆,李赟卉,等.转基因vp28蓝藻口服疫苗半数有效量测定及其对斑马鱼的安全评价[J].水产学报,2021,45(2):255-264.
16	谢雪晴,田钰琪,田敬欢,等.T7 RNA聚合酶基因表达系统在鱼腥藻7120中构建及hG-CSF的表达[J].生物工程学报,2020,36(11):2467-2477.
17	朱婵,施定基,何培民,等.转基因聚球藻7942的vp19基因表达效率及其光合生理特性研究[J].上海农业学报,2021,37(4): 36-41.
18	朱小明,章军,徐虹,等.转胸腺素基因螺旋藻的表达及免疫增强活性研究[J].福建农业学报,2005,20(4):228-232.
19	LUAN G, LU X. Tailoring cyanobacterial cell factory for improved industrial properties[J]. Biotechnol. Adv., 2018, 36(2): 430-442.
20	XUE Y, HE Q. Cyanobacteria as cell factories to produce plant secondary metabolites[J/OL]. Front. Bioeng. Biotechnol., 2015, 3: 57[2022-12-20]. .
21	DESHPANDE A, VUE J, MORGAN J. Combining random mutagenesis and metabolic engineering for enhanced tryptophan production in Synechocystis sp. strain PCC6803[J/OL]. Appl. Environ, Microbiol., 2020, 86(9): e02816-19[2022-12-20]. .
22	YU R, YAMADA A, WATANABE K, et al.. Production of eicosapentaenoic acid by a recombinant marine cyanobacterium, Synechococcus sp.[J]. Lipids, 2000, 35(10): 1061-1064.
23	SANTOS-MERINO M, GARCILLÁN-BARCIA M P, DE L A CRUZ F. Engineering the fatty acid synthesis pathway in Synechococcus elongates PCC 7942 improves omega-3 fatty acid production[J/OL]. Biotechnol. Biofuels., 2018, 11: 239[2022-12-20]. .
24	LIN P C, SAHA R, ZHANG F, et al.. Metabolic engineering of the pentose phosphate pathway for enhanced limonene production in the cyanobacterium Synechocystis sp. PCC6803[J/OL]. Sci. Rep., 2017, 7(1): 17503[2022-12-20]. .
25	ENGLUND E, PATTANAIK B, UBHAYASEKERA S J, et al.. Production of squalene in Synechocystis sp. PCC 6803[J/OL]. PLoS ONE, 2014, 9(3): e90270[2022-12-20]. .
26	WANG X, CHEN L, LIU J, et al.. Light-driven biosynthesis of myo-inositol directly from CO2 in Synechocystis sp. PCC6803[J/OL]. Front. Microbiol., 2020, 11: 566117[2022-12-20]. .
27	DIAO J, SONG X, ZHANG L, et al.. Tailoring cyanobacteria as a new platform for highly efficient synthesis of astaxanthin[J]. Metab. Eng., 2020, 61: 275-287.
28	KACHEL B, MACK M. Engineering of Synechococcus sp. strain PCC7002 for the photoautotrophic production of light-sensitive riboflavin (vitamin B2)[J]. Metab. Eng., 2020, 62: 275-286.
29	FAN E S, LU K W, WEN R C, et al.. Photosynthetic reduction of xylose to xylitol using cyanobacteria[J/OL]. Biotechnol. J., 2020, 15(6): e1900354[2022-12-20]. .
30	QIAO Y, WANG W, LU X. Engineering cyanobacteria as cell factories for direct trehalose production from CO₂ [J]. Metab. Eng., 2020, 62: 161-171.
31	ANDREWS F, FAULKNER M, TOOGOOD H S, et al.. Combinatorial use of environmental stresses and genetic engineering to increase ethanol titres in cyanobacteria[J/OL]. Biotechnol. Biofuels., 2021, 14(1): 240[2022-12-20]. .
32	AGARWAL P, SONI R, KAUR P, et al.. Cyanobacteria as a promising alternative for sustainable environment: synthesis of biofuel and biodegradable plastics[J/OL]. Front. Microbiol., 2022, 13: 939347[2022-12-20]. .
33	VEETIL V P, ANGERMAYR S A, HELLINGWERF K J. Ethylene production with engineered Synechocystis sp. PCC6803 strains[J/OL]. Microb. Cell. Fact., 2017, 16(1): 34[2022-12-20]. .
34	VARMAN A M, XIAO Y, PAKRASI H B, et al.. Metabolic engineering of Synechocystis sp. strain PCC6803 for isobutanol production[J]. Appl. Environ. Microbiol., 2013, 79(3): 908-914.
35	OLIVER J W, MACHADO I M, YONEDA H, et al.. Cyanobacterial conversion of carbon dioxide to 2,3-butanediol[J]. Proc. Natl Acad. Sci. USA, 2013, 110(4): 1249-1254.
36	TAN C, TAO F, XU P. Direct carbon capture for the production of high-performance biodegradable plastics by cyanobacterial cell factories[J]. Green. Chem., 2022, 24: 4470-4483.
37	张春月,金佳杨,邱勇隽,等.传统与未来的碰撞:食品发酵工程技术与应用进展[J].生物技术进展,2021,11(4):418-429.
38	LIU D, LIBERTON M, HENDRY J I, et al.. Engineering biology approaches for food and nutrient production by cyanobacteria[J]. Curr. Opin. Biotechnol., 2021, 67: 1-6.
39	KRISHNAN A, QIAN X, ANANYEV G, et al.. Rewiring of cyanobacterial metabolism for hydrogen production: synthetic biology approaches and challenges[J]. Adv. Exp. Med. Biol., 2018, 1080: 171-213.
40	VELMURUGAN R, INCHAROENSAKDI A. Metabolic transformation of cyanobacteria for biofuel production[J/OL]. Chemosphere, 2022, 299: 134342[2022-12-20]. .
41	REDDING K E, APPEL J, BOEHM M, et al.. Advances and challenges in photosynthetic hydrogen production[J]. Trends. Biotechnol., 2022, 40(11): 1313-1325.
42	WANG F, GAO Y, YANG G. Recent advances in synthetic biology of cyanobacteria for improved chemicals production[J]. Bioengineered, 2020, 11(1): 1208-1220.
43	LI C, ZHENG J, WU Y, et al.. Light-driven synthetic biology: progress in research and industrialization of cyanobacterial cell factory[J/OL]. Life(Basel), 2022, 12(10): 1537[2022-12-20]. .
44	SENGUPTA A, PAKRASI H B, WANGIKAR P P. Recent advances in synthetic biology of cyanobacteria[J]. Appl. Microbiol. Biotechnol., 2018, 102(13): 5457-5471.
45	ENGLUND E, LIANG F, LINDBERG P. Evaluation of promoters and ribosome binding sites for biotechnological applications in the unicellular cyanobacterium Synechocystis sp. PCC6803[J/OL]. Sci. Rep., 2016, 6: 36640[2022-12-20]. .
46	THIEL K, MULAKU E, DANDAPANI H, et al.. Translation efficiency of heterologous proteins is significantly affected by the genetic context of RBS sequences in engineered cyanobacterium Synechocystis sp. PCC6803[J/OL]. Microb. Cell. Fact, 2018, 17(1): 34[2022-12-20]. .
47	LIU D, PAKRASI H B. Exploring native genetic elements as plug-in tools for synthetic biology in the cyanobacterium Synechocystis sp. PCC6803[J/OL]. Microb. Cell. Fact, 2018, 17(1): 48[2022-12-20]. .
48	曹豪豪,张红兵,薛溪发,等.新型基因编辑技术在单细胞微藻中的应用进展[J].生物技术进展,2021,11(1):9-15.
49	BALDANTA S, GUEVARA G, NAVARRO-LLORENS J M. SEVA-Cpf 1, a CRISPR-Cas12a vector for genome editing in cyanobacteria[J/OL]. Microb. Cell. Fact, 2022, 21(1): 103[2022-12-20]. .
50	UNGERER J, PAKRASI H B. Cpf1 is a versatile tool for CRISPR genome editing across diverse species of cyanobacteria[J/OL]. Sci. Rep., 2016, 6: 39681[2022-12-20]. .
51	GORDON G C, KOROSH T C, CAMERON J C, et al.. CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC7002[J]. Metab. Eng., 2016, 38: 170-179.
52	YAO L, SHABESTARY K, BJÖRK S M, et al.. Pooled CRISPRi screening of the cyanobacterium Synechocystis sp. PCC6803 for enhanced industrial phenotypes[J/OL]. Nat. Commun., 2020, 11(1): 1666[2020-04-03]. .
53	HITCHCOCK A, HUNTER C N, CANNIFFE D P. Progress and challenges in engineering cyanobacteria as chassis for light-driven biotechnology[J]. Microb. Biotechnol., 2020, 13(2): 363-367.
54	钱美文,谭春林,倪俊,等.蓝细菌细胞工厂合成聚合物单体的研究进展[J].生物工程学报,2021,37(3):1017-1031.
55	TREECE T R, GONZALES J N, PRESSLEY J R, et al.. Synthetic biology approaches for improving chemical production in cyanobacteria[J/OL]. Front. Bioeng. Biotechnol., 2022, 10: 869195[2022-12-20]. .

外源蛋白	功能	藻株	文献
人肝金属硫蛋白-I_A（hMT-I_A）	金属代谢、抗氧化等	Anabaena PCC7120	[6]
人尿激酶原基因(pro-urokinase)	溶栓	Synechococcus sp. PCC7002	[7]
人肿瘤坏死因子α（hTNF-α）	杀伤肿瘤细胞	Anabaena PCC7102	[8]
人表皮生长因子（hEGF）	促进表皮细胞生长等	Synechococcus sp. PCC7002	[9]
人铜锌超氧化物歧化酶（hCu，Zn-SOD）	抗氧化、抗辐射损伤等	Synechococcus sp. PCC7942	[10]
人粒-巨噬细胞集落刺激因子（hGM-CSF）)	造血调控、免疫调节等	Anabaena PCC7102	[11]
Eta1-L-Gapdh蛋白	预防鱼类迟缓爱德华氏菌感染	Anabaena PCC7120	[12]
病毒包膜蛋白（vp28）	预防对虾白斑综合征病毒	Synechococcus sp. PCC7942, Anabaena PCC7120	[13-15]
人粒细胞集落刺激因子（hG-CSF）	造血调控	Anabaena PCC7102	[16]
病毒包膜蛋白（vp19）	预防对虾白斑综合征病毒	Synechococcus sp. PCC7942	[17]
胸腺α₁（Tα₁）	增强免疫	Spirulina	[18]

外源蛋白	功能	藻株	文献
人肝金属硫蛋白-I_A（hMT-I_A）	金属代谢、抗氧化等	Anabaena PCC7120	[6]
人尿激酶原基因(pro-urokinase)	溶栓	Synechococcus sp. PCC7002	[7]
人肿瘤坏死因子α（hTNF-α）	杀伤肿瘤细胞	Anabaena PCC7102	[8]
人表皮生长因子（hEGF）	促进表皮细胞生长等	Synechococcus sp. PCC7002	[9]
人铜锌超氧化物歧化酶（hCu，Zn-SOD）	抗氧化、抗辐射损伤等	Synechococcus sp. PCC7942	[10]
人粒-巨噬细胞集落刺激因子（hGM-CSF）)	造血调控、免疫调节等	Anabaena PCC7102	[11]
Eta1-L-Gapdh蛋白	预防鱼类迟缓爱德华氏菌感染	Anabaena PCC7120	[12]
病毒包膜蛋白（vp28）	预防对虾白斑综合征病毒	Synechococcus sp. PCC7942, Anabaena PCC7120	[13-15]
人粒细胞集落刺激因子（hG-CSF）	造血调控	Anabaena PCC7102	[16]
病毒包膜蛋白（vp19）	预防对虾白斑综合征病毒	Synechococcus sp. PCC7942	[17]
胸腺α₁（Tα₁）	增强免疫	Spirulina	[18]

产物	功能	藻种名	文献
色氨酸	动物饲料添加剂等	Synechocystis sp. PCC6803	[21]
二十碳五烯酸	降血脂、降血压等	Synechococcus sp. NKBG042902	[22]
ω-3脂肪酸	调整血脂、生物能源	Synechococcus sp. NKBG15041c	[23]
柠檬烯	镇咳、祛痰、抑菌等	Synechocystis sp. PCC6803	[24]
鲨烯	药用	Synechocystis sp. PCC6803	[25]
肌醇	生长因子	Synechocystis sp. PCC6803	[26]
虾青素	抗氧化	Synechocystis sp. PCC6803	[27]
维生素B2	维生素	Synechococcus sp. PCC7002	[28]
木糖醇	防龋齿、改善肝功能、调节肠道等	Synechococcus sp. PCC7942	[29]
海澡糖	食品添加剂	Synechocystis sp. PCC6803	[30]
乙醇	生物能源	Synechococcus sp. PCC7942, Synechococcus sp. PCC7002	[31]
氢气	生物能源	Cyanothece sp. ATCC51142	[32]
聚羟基脂肪酸酯	生物塑料	Synechococcus sp. MA19	[32]
乙烯	化工原料	Synechocystis sp. PCC6803	[33]
异丁醇	化工原料、能源	Synechocystis sp. PCC6803	[34]
丁二醇	化工原料、能源	Synechococcus sp. PCC7942	[35]
聚乳酸	生物塑料	Synechococcus sp. PCC7942	[36]

产物	功能	藻种名	文献
色氨酸	动物饲料添加剂等	Synechocystis sp. PCC6803	[21]
二十碳五烯酸	降血脂、降血压等	Synechococcus sp. NKBG042902	[22]
ω-3脂肪酸	调整血脂、生物能源	Synechococcus sp. NKBG15041c	[23]
柠檬烯	镇咳、祛痰、抑菌等	Synechocystis sp. PCC6803	[24]
鲨烯	药用	Synechocystis sp. PCC6803	[25]
肌醇	生长因子	Synechocystis sp. PCC6803	[26]
虾青素	抗氧化	Synechocystis sp. PCC6803	[27]
维生素B2	维生素	Synechococcus sp. PCC7002	[28]
木糖醇	防龋齿、改善肝功能、调节肠道等	Synechococcus sp. PCC7942	[29]
海澡糖	食品添加剂	Synechocystis sp. PCC6803	[30]
乙醇	生物能源	Synechococcus sp. PCC7942, Synechococcus sp. PCC7002	[31]
氢气	生物能源	Cyanothece sp. ATCC51142	[32]
聚羟基脂肪酸酯	生物塑料	Synechococcus sp. MA19	[32]
乙烯	化工原料	Synechocystis sp. PCC6803	[33]
异丁醇	化工原料、能源	Synechocystis sp. PCC6803	[34]
丁二醇	化工原料、能源	Synechococcus sp. PCC7942	[35]
聚乳酸	生物塑料	Synechococcus sp. PCC7942	[36]

[1]	陈洁, 黄永康, 王希. 合成生物学在化工新材料领域的应用及展望[J]. 生物技术进展, 2023, 13(1): 39-45.
[2]	王安琪,朱华新,赵翔,崔建霞,王琰,崔海信. 基于纳米基因载体的动植物遗传转化研究进展[J]. 生物技术进展, 2018, 8(4): 293-301.
[3]	张云威,苏航,刁勇,戚智青. 牛乳铁蛋白肽的抑菌功能及基因工程制备进展[J]. 生物技术进展, 2016, 6(4): 235-238.
[4]	李天丽,郑凌凌,张琪,宋立荣. 蓝藻无菌化方法研究进展[J]. 生物技术进展, 2015, 5(5): 329-334.
[5]	邱庆庆,任秀莲,吴泽,陈泳兴,魏琦峰. 海藻生物质能源转化的研究现状[J]. 生物技术进展, 2015, 5(3): 153-157.
[6]	邓龙,周思,郭新东. 基于基因突变的基因工程抗体亲和力成熟研究[J]. 生物技术进展, 2014, 4(6): 400-404.
[7]	何明雄,吴波,谭芙蓉,王景丽,税宗霞,秦晗,代立春,胡启春,. 运动发酵单胞菌在生物炼制中的研究进展[J]. 生物技术进展, 2014, 4(5): 331-339.
[8]	牛丽芳,路铁刚,林浩. 水稻高光效育种研究进展[J]. 生物技术进展, 2014, 4(3): 153-157.
[9]	杨瑞红. 生物脱硫微生物及基因工程应用的研究进展[J]. 生物技术进展, 2013, 3(3): 190-195.
[10]	张亮, 张兰, 王磊. 植物维生素E基因工程研究进展[J]. 生物技术进展, 2012, 2(6): 397-403.
[11]	袁志勇,朱红惠,霍光华,冯广达. 微囊藻毒素生物合成及其检测的分子生物学研究进展[J]. 生物技术进展, 2012, 2(5): 328-334.
[12]	苗猛猛,王旭静,唐巧玲,王志兴. 抗蚜基因工程研究进展[J]. 生物技术进展, 2012, 2(2): 104-109.
[13]	王国增,李轶女,张志芳,沈桂芳. 除草剂抗性基因的研究进展[J]. 生物技术进展, 2011, 1(6): 398-402.

蓝藻细胞工厂的研究进展

Research Progress of Cyanobacteria Cell Factories

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 2

参考文献 55

相关文章 13

编辑推荐

Metrics

本文评价