DESCRIPTION
Background & Inspiration
The ocean, including various habitats and environmental conditions from shallow water to deep water, breeds a large number of animals, plants, and microbial communities with extremely high diversity, resulting in a large number of bioactive secondary metabolites, forming a treasure trove of biological and resource diversity that has not yet been developed and underutilized. The deep sea refers to the ocean below 1000 meters in depth[1]. It is one of the most extensive and least explored habitats on the earth and an important part of the construction of a maritime power. As an extreme ecological environment, there are also deep-sea microorganisms with special diversity and high novelty in the deep sea, which may produce various special bioactive secondary metabolites through different biosynthetic gene clusters as environmental adaptation mechanisms. Therefore, the resource development of microbial secondary metabolites in deep-sea extreme environments and the demonstration of the uniqueness and diversity of secondary metabolites and biosynthetic gene clusters can improve the understanding of deep-sea microbial functions and product diversity[2]. It is very important to understand the adaptive evolution of microorganisms to extreme environments, the formation and maintenance of deep-sea biodiversity, and the function of deep-sea ecosystems, thus providing a research basis for the sustainability and ecological protection of marine ecosystems[3].
Fig1.The zonation of the ocean and its depth[5]
Terpene secondary metabolites are structurally diverse functional compounds found in plants, fungi and bacteria. They have a variety of biological activities such as antibacterial, anti-inflammatory, anti-oxidation, anti-tumor and neuroprotective effects, and have broad application prospects in various fields such as medicine and food. At present, the secondary metabolites of commercial terpenes are mainly derived from traditional plant or fungal extracts. However, due to its slow growth, limited arable land resources, complex and lengthy extraction process and other factors, the key functional terpene secondary metabolites are difficult to excavate and purify. Therefore, we need a wider source of secondary metabolites and a more efficient way to obtain secondary metabolites. Microbial cell factories have shown great potential in the efficient synthesis of secondary metabolites. Through genetic engineering technology, microorganisms can efficiently produce specific secondary metabolites under controlled conditions, which can be used in the fields of drugs, skin care raw materials and food additives[4]. We speculate that deep-sea microorganisms have great potential for biosynthesis, and the terpenoid secondary metabolites produced have broad application prospects.
Fig2. Some novel antimicrobial secondary metabolites from marine bacteria [6]
This project will integrate metagenomic samples from deep-sea ecosystems for data analysis, construct a set of deep-sea microbial genomes and a set of biosynthetic gene clusters, systematically analyze and sort out the genetic resources of secondary metabolites, and directionally explore new terpenes with high-value biological activity. Secondary metabolites and their biosynthetic gene clusters, optimization and direct cloning to obtain 1-2 terpene BGCs. The high-throughput biosynthesis system was used to realize the synthesis and expression of 1-2 terpene secondary metabolites and analyze the structure of the products. It is expected to reveal the biosynthesis mechanism of 1-2 new terpene secondary metabolites with high value biological activity in deep-sea metagenome, which can be used to evaluate their biological activities such as antibacterial, anti-inflammatory, anti-tumor and neuroprotective effects.
Design
This project will integrate metagenomic samples from deep-sea ecosystems for data analysis, construct a set of deep-sea microbial genomes and a set of biosynthetic gene clusters, systematically analyze and sort out the genetic resources of secondary metabolites, and directionally explore new terpene secondary metabolites with high value biological activity and their biosynthetic gene clusters. Two terpene BGCs were obtained by optimization and direct cloning. Using a high-throughput biosynthesis system, we achieved the synthesis and expression of two terpene secondary metabolites and attempted to analyze the product structure. Specifically, we established two metabolic pathways in E.coli BL21, and produced new terpene secondary metabolites by constructing recombinant plasmids, transforming BL21, and fermentation.
Fig3. Experimental design route
Goal
This project aims to excavate terpene secondary metabolites and construct a high-throughput microbial heterologous expression platform for new terpene secondary metabolites with high value biological activity, which can further expand the biosynthesis research content of terpene secondary metabolites and provide important scientific basis for efficient biosynthesis and synthesis process optimization of terpene secondary metabolites. The development of secondary metabolites of microbial terpenes in deep-sea extreme environments can expand the application of secondary metabolites derived from deep-sea microorganisms in medicine, food and other fields. On the other hand, it can also further promote the close integration and innovative development of biotechnology, medicine, food and other industries, bring new products and solutions, and provide safe, effective and sustainable natural ingredients.
Reference
[1] Alma'abadi AD, Gojobori T, Mineta K. Marine Metagenome as A Resource for Novel Enzymes. Genomics Proteomics Bioinformatics. 2015 Oct;13(5):290-5. doi: 10.1016/j.gpb.2015.10.001. Epub 2015 Nov 10. PMID: 26563467; PMCID: PMC4678775.
[2] Sunagawa S, Coelho LP, Chaffron S, Kultima JR, Labadie K, Salazar G, Djahanschiri B, Zeller G, Mende DR, Alberti A, Cornejo-Castillo FM, Costea PI, Cruaud C, d'Ovidio F, Engelen S, Ferrera I, Gasol JM, Guidi L, Hildebrand F, Kokoszka F, Lepoivre C, Lima-Mendez G, Poulain J, Poulos BT, Royo-Llonch M, Sarmento H, Vieira-Silva S, Dimier C, Picheral M, Searson S, Kandels-Lewis S; Tara Oceans coordinators; Bowler C, de Vargas C, Gorsky G, Grimsley N, Hingamp P, Iudicone D, Jaillon O, Not F, Ogata H, Pesant S, Speich S, Stemmann L, Sullivan MB, Weissenbach J, Wincker P, Karsenti E, Raes J, Acinas SG, Bork P. Ocean plankton. Structure and function of the global ocean microbiome. Science. 2015 May 22;348(6237):1261359. doi: 10.1126/science.1261359. PMID: 25999513.
[3] Sunagawa S, Coelho LP, Chaffron S, Kultima JR, Labadie K, Salazar G, Djahanschiri B, Zeller G, Mende DR, Alberti A, Cornejo-Castillo FM, Costea PI, Cruaud C, d'Ovidio F, Engelen S, Ferrera I, Gasol JM, Guidi L, Hildebrand F, Kokoszka F, Lepoivre C, Lima-Mendez G, Poulain J, Poulos BT, Royo-Llonch M, Sarmento H, Vieira-Silva S, Dimier C, Picheral M, Searson S, Kandels-Lewis S; Tara Oceans coordinators; Bowler C, de Vargas C, Gorsky G, Grimsley N, Hingamp P, Iudicone D, Jaillon O, Not F, Ogata H, Pesant S, Speich S, Stemmann L, Sullivan MB, Weissenbach J, Wincker P, Karsenti E, Raes J, Acinas SG, Bork P. Ocean plankton. Structure and function of the global ocean microbiome. Science. 2015 May 22;348(6237):1261359. doi: 10.1126/science.1261359. PMID: 25999513.
[4] Schempp FM, Drummond L, Buchhaupt M, Schrader J. Microbial Cell Factories for the Production of Terpenoid Flavor and Fragrance Compounds. J Agric Food Chem. 2018 Mar 14;66(10):2247-2258. doi: 10.1021/acs.jafc.7b00473. Epub 2017 Apr 18. PMID: 28418659
[5]Britannica, The Editors of Encyclopaedia. "bathyal zone". Encyclopedia Britannica, 27 Jan. 2015, https://www.britannica.com/science/bathyal-zone. Accessed 29 September 2024.
[6] Srinivasan, R.; Kannappan, A.; Shi, C.; Lin, X. Marine Bacterial Secondary Metabolites: A Treasure House for Structurally Unique and Effective Antimicrobial Compounds. Mar. Drugs 2021, 19, 530. https://doi.org/10.3390/md19100530