工业丝状真菌土曲霉合成生物技术研究进展及展望

doi:10.12211/2096-8280.2020-002

摘要/Abstract

摘要：

丝状真菌作为一类重要的微生物，在食品、医药、化工等国计民生领域发挥了重要作用，其合成生物技术的发展已经展示了更大的工业应用潜力。土曲霉是一种非常具有工业生产价值的丝状真菌，是生物基化学品衣康酸和降血脂药物洛伐他汀的工业生产菌，展现出了强大的天然产物合成能力和突出的工业发酵性能，具有成为典型性丝状真菌底盘细胞应用于合成生物技术开发的价值。近年来，在土曲霉工业菌株改造与发酵工艺优化、生物合成机制解析等方面都开展了系列的研究，显著推动了其合成生物技术研究的发展。本文综述了土曲霉合成生物技术工具开发、衣康酸和洛伐他汀工业生产菌株改造优化、次级代谢产物生物合成机制解析等方面的最新研究进展，并对土曲霉的合成生物技术应用前景和发展方向进行了展望。挖掘土曲霉底盘细胞在合成生物学研究中的优势，可以更好地拓展合成生物技术的应用。

关键词: 丝状真菌, 土曲霉, 合成生物学, 细胞工厂, 洛伐他汀, 衣康酸, 天然产物生物合成

Abstract:

Filamentous fungi are a class of important microorganisms, which have played an important role in the fields of industrial sectors and people's daily life for the production of food, medicine, and chemical products. With the development of synthetic biology, filamentous fungi have shown more potentials. Aspergillus terreus is an industrially valuable filamentous fungus, which has been applied in the production of the bio-based chemical itaconic acid and the lipid-lowering drug lovastatin. The excellent synthetic ability of natural products and outstanding fermentation properties of A. terreus have been fully demonstrated through industrial applications. Meanwhile, it has also been acknowledged that A. terreus would be a promising filamentous fungal chassis cell for synthetic biology development. In recent years, many studies have been performed using A. terreus, including genetic engineering of industrial strains, optimization of fermentation processes, elucidation of biosynthetic mechanisms, etc., which have significantly contributed to the development of A. terreus synthetic biology. Herewith, these recent research advances are reviewed from: 1) Synthetic biology tools for A. terreus. The development of genetic operation systems is introduced, including the characterization and application of promoters and markers and the progress of gene-editing technologies. 2) Industrial cell factory for the lipid-lowering drug lovastatin. In addition to the biosynthetic pathway of lovastatin, the design and construction of an efficient monacolin J-producing cell factory based on the industrial A. terreus strains are introduced in details. 3) Industrial cell factory for the bio-based chemical itaconic acid. The biosynthetic pathway of itaconic acid and the systematic metabolic engineering of the industrial strain are introduced. 4) Biosynthesis of secondary metabolites. The biosynthetic mechanism and research strategies for the secondary metabolites are introduced. 5) The advantages and prospects of A. terreus chassis cells in synthetic biology are summarized, and the practical applications are listed. Based on the research progress of A. terreus and other filamentous fungi, the challenges and future work of A. terreus biosynthesis technology development are proposed, including enriching the genetic element library, developing efficient gene editing methods and building high-throughput evaluation platforms. Addressing these challenges will provide more alternative design strategies for building more efficient cell factories of A. terreus, and will also contribute to promoting synthetic biology research of filamentous fungi.

Key words: Filamentous fungi, Aspergillus terreus, synthetic biology, cell factory, lovastatin, itaconic acid, biosynthesis of natural products

中图分类号:

Q939.97

黄雪年, 唐慎, 吕雪峰. 工业丝状真菌土曲霉合成生物技术研究进展及展望[J]. 合成生物学, 2020, 1(2): 187-211.

HUANG Xuenian, TANG Shen, LV Xuefeng. Progress and prospect for synthetic biology research of the industrial filamentous fungi Aspergillus terreus[J]. Synthetic Biology Journal, 2020, 1(2): 187-211.

图/表 16

图1 土曲霉的各种形态

Fig. 1 The morphologies of A. terreus

图2 土曲霉洛伐他汀生物合成途径

Fig. 2 Biosynthesis pathway of lovastain in A. terreus

图3 生产辛伐他汀前体物monacolin J土曲霉细胞工厂的设计策略

Fig. 3 Construction of monacolin J-producing cell factory in A.terreus

图4 土曲霉的衣康酸生物合成途径

Fig. 4 The biosynthesis pathway of itaconic acid in A. terreus

图5 产衣康酸土曲霉菌株改造策略A. terreus

Fig. 5 Metabolic engineering of the itaconic acid-producing

表1 土曲霉已被研究基因簇及其下游产物

Tab. 1 Biosynthetic gene clusters and products identified in A. terreus

序号	核心基因编号	类型	代表性产物	参考文献
1	ATEG_00145 (terA)	nrPKS	terrein	[82,83]
2	ATEG_03432	nrPKS	6-acetyl-2,7-dihydroxy-3-methylnaphthalene-1,4-dione	[81]
3	ATEG_06275 (atX)	hrPKS	terreic acid	[84,85]
4	ATEG_08451 (gedC)	nrPKS	geodin	[86,87]
5	ATEG_10080 (trt4)	nrPKS	terretonin	[88,89]
6	ATEG_09617 (ctvA)	hrPKS	citreoviridin	[90]
7	ATEG_09961 (lovB)	hrPKS	lovastatin	[24]
7	ATEG_09968 (lovF)	hrPKS	lovastatin	[24]
8	Atcurs1	hrPKS	10,11-dehydrocurvularin	[91]
8	Atcurs2	nrPKS	10,11-dehydrocurvularin	[91]
9	ATEG_03629	nrPKS	azasperpyranone A	[92]
	ATEG_03630	NRPS-like
	ATEG_07659	hrPKS
	ATEG_07661	nrPKS
10	ATEG_03470 (ataP)	NRPS	acetylaranotin	[93]
11	ATEG_10305 (anaPS)	NRPS	asterrelenin	[2]
12	ATEG_09064 (apmB)	NRPS	asperphenamate	[94]
12	ATEG_09068 (apmA)	NRPS	asperphenamate	[94]
13	ATEG_00700 (atqA)	NRPS-like	asterriquinones CT5	[2]
14	ATEG_02004 (apvA)	NRPS-like	aspulvinones B1	[2]
15	ATEG_02815 (btyA)	NRPS-like	butyrolactones I	[95]
16	ATEG_03563 (atmelA)	NRPS-like	aspulvinone E (asp-melanin前体)	[95]
17	ATEG_08899 (pgnA)	NRPS-like	phenguignardic acid	[7]
18	ATEG_00325	PKS-NRPS	isoflavipucine	[96]
19	ATEG_00913 (pytA)	PKS-NRPS	pyranterreone C	[76]
20	ATEG_04416 (astA)	TC	aspterric acid	[97]

图6 (+)-Terrein的生物合成基因簇及合成途径

Fig. 6 Gene cluster in the biosynthesis pathway of (+)-terrein in A. terreus

图7 Terreic acid的生物合成基因簇及合成途径

Fig. 7 Gene cluster in the biosynthesis pathway of terreic acid in A. terreus

图8 Citreoviridin的生物合成基因簇及合成途径

Fig. 8 Gene cluster in the biosynthesis pathway of citreoviridin in A. terreus

图9 Acetylaranotin的生物合成基因簇及合成途径

Fig. 9 Gene cluster in the biosynthesis pathway of acetylaranotin in A. terreus

图10 土曲霉Asp类黑色素的生物合成机制

Fig. 10 Biosynthesis of Asp-melanin in A. terreus

图11 Aspterric acid的生物合成基因簇及合成途径

Fig. 11 Gene cluster in the biosynthesis pathway of aspterric acid in A. terreus

图12 Terretonin的生物合成基因簇及合成途径

Fig. 12 Gene cluster in the biosynthesis pathway of terretonin acid

图13 Isoflavipucine的生物合成基因簇及合成途径

Fig. 13 Gene cluster in the biosynthesis pathway of isoflavipucine in A. terreus

图14 Pyranterreones的生物合成基因簇及途径

Fig. 14 Gene cluster in the biosynthesis pathway of pyranterreones in A. terreus

图15 两个独立基因簇协同合成化合物azasperpyranones

Fig. 15 Synergistic synthesis of azasperpyranones under the catalysis of key enzymes encoded by two gene clusters

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