17-ERACoBioTech: MicroalgaE as Renewable Innovative green cell facTories

17-ERACoBioTech：微藻作为可再生创新绿色细胞工厂

• 批准号: BB/R021694/1
• 负责人: Alison Smith
• 金额: $ 47.61万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR021694%2F1
• 关键词: 17; ERACoBioTech; MicroalgaE; Renewable; Innovative

This project [MERIT] sets out to leverage state of the art techniques to develop microalgae for the sustainable production of high-value, medically and industrially relevant molecules from carbon dioxide and light. Specifically, MERIT will focus on the production of twenty carbon containing (C20) molecules, known as diterpenoids. These are complex metabolites found largely in plants, and whose variety and complexity have made them incredibly interesting for numerous applications such as medicines, antimicrobial agents, and high-value chemicals. The complex structures of diterpenoids are difficult and costly to synthesize chemically and can be expensive or inefficient to purify from their native host organisms.All organisms produce the same 5-carbon building blocks used in diterpenoid production pathways. By introducing these production pathways between species, we can produce non-native diterpenoids in engineered hosts (e.g. bacteria, yeast or microalgae). Production of diterpenoid products in fermentative hosts (e.g. yeast) has become a mature technology, however, relies on unsustainable use of organic carbon sources such as glucose and inherently competes for agricultural resources. Microalgae, however, are naturally optimized to produce precursor molecules needed to make diterpenoids, as these same molecules are used for harvesting light and for generation of antioxidant pigments in the cell. Algae hold the additional benefit of rapid growth rates in simple mineral salt solutions using only light and CO2 as energy inputs. These organisms are ideal hosts for the production of diterpenes and are inherently sustainable production chassis.By introducing different enzymes that convert precursor molecules into the numerous carbon skeletons of diterpenoid products algal cells can be engineered to serve as diterpenoid production hosts. Additional engineering of these algal cells through introduction of enzymes that add oxygen molecules into the diterpene backbones will allow the algae to produce specialized diterpene structures, which often have medicinal properties. To date, engineering of microalgae to accumulate new non-native enzymes has been a major limiting factor to their widespread application as green-cell factories for complex biotechnological targets. Combining strain domestication with synthetic biology (an emerging discipline that uses engineering principles to design and assemble biological components) has recently been demonstrated to facilitate advanced engineering of these highly promising organisms. In this project, an international team representing global leaders in algal synthetic biology, outdoor algal cultivation, photobioreactor design, and process modelling, will join forces to drive forward the development of algal cell factories. The MERIT team already successfully engineered pathways for the production of several diterpenoids in microalgae. Multiple levels of strain engineering and synthetic biology will be implemented to create green-cell factories with enhanced carbon flow from CO2 to diterpenoids. Various enzymes will be combined to produce novel 'new-to-nature' diterpenoid products with potential for numerous applications. Optimized strains will be grown to scale and processes for extraction of the products will be designed. The project will generate many new avenues of commercialization potential and significantly contribute to the development of the European Bioeconomy.

该项目 [MERIT] 旨在利用最先进的技术开发微藻，以利用二氧化碳和光可持续生产高价值、医学和工业相关的分子。具体来说，MERIT 将专注于生产二十种含碳 (C20) 分子，即二萜类化合物。这些是主要存在于植物中的复杂代谢物，其多样性和复杂性使其在药物、抗菌剂和高价值化学品等众多应用中具有极大的吸引力。二萜类化合物结构复杂，化学合成难度大且成本高，从其天然宿主生物体中纯化也可能成本高昂或效率低下。所有生物体都产生用于二萜类化合物生产途径的相同 5 碳结构单元。通过在物种之间引入这些生产途径，我们可以在工程宿主（例如细菌、酵母或微藻）中生产非天然二萜类化合物。在发酵宿主（例如酵母）中生产二萜产品已成为一项成熟的技术，然而，依赖于葡萄糖等有机碳源的不可持续利用，并且本质上会争夺农业资源。然而，微藻经过天然优化，可以产生制造二萜类化合物所需的前体分子，因为这些相同的分子用于收集光并在细胞中产生抗氧化色素。藻类在仅使用光和二氧化碳作为能量输入的简单矿物盐溶液中具有快速生长的额外优势。这些生物体是生产二萜的理想宿主，并且本质上是可持续的生产底盘。通过引入将前体分子转化为二萜产品的众多碳骨架的不同酶，可以将藻类细胞改造为二萜生产宿主。通过引入酶将氧分子添加到二萜主链中，对这些藻类细胞进行额外的工程改造，将使藻类产生专门的二萜结构，这些结构通常具有药用特性。迄今为止，微藻工程积累新的非天然酶一直是其作为复杂生物技术目标的绿色细胞工厂广泛应用的主要限制因素。最近证明，将菌株驯化与合成生物学（一门利用工程原理设计和组装生物成分的新兴学科）相结合，可以促进这些极具前景的生物体的先进工程。在该项目中，代表藻类合成生物学、户外藻类培养、光生物反应器设计和过程建模领域全球领导者的国际团队将联手推动藻类细胞工厂的发展。 MERIT 团队已经成功设计了在微藻中生产多种二萜类化合物的途径。将实施多层次的菌株工程和合成生物学，以创建绿色细胞工厂，增强从二氧化碳到二萜类化合物的碳流。各种酶将被组合起来生产具有多种应用潜力的新型“新自然”二萜类产品。优化的菌株将按规模生长，并设计产品的提取工艺。该项目将产生许多具有商业化潜力的新途径，并对欧洲生物经济的发展做出重大贡献。

项目成果

Thiamine metabolism genes in diatoms are not regulated by thiamine despite the presence of predicted riboswitches.

尽管存在预测的核糖开关，但硅藻中的硫胺素代谢基因不受硫胺素调节。

• DOI: http://dx.10.17863/cam.86078
• 发表时间: 2022
• 作者: Llavero

Exploring the Impact of Terminators on Transgene Expression in Chlamydomonas reinhardtii with a Synthetic Biology Approach.

用合成生物学方法探索终止子对莱茵衣藻转基因表达的影响。

• DOI: http://dx.10.17863/cam.76329
• 发表时间: 2021
• 作者: Geisler K

Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii.

光合底盘的诞生：MoClo 工具包实现微藻莱茵衣藻的合成生物学。

• DOI: http://dx.10.17863/cam.32537
• 发表时间: 2018
• 作者: Crozet P

The Algal Chloroplast as a Testbed for Synthetic Biology Designs Aimed at Radically Rewiring Plant Metabolism.

藻类叶绿体作为合成生物学设计的试验台，旨在从根本上重新布线植物代谢。

• DOI: http://dx.10.3389/fpls.2021.708370
• 发表时间: 2021
• 期刊: Frontiers in plant science
• 作者: Jackson HO

Exploring the impact of terminators on transgene expression in Chlamydomonas reinhardtii with a synthetic biology approach

用合成生物学方法探索终止子对莱茵衣藻转基因表达的影响

• DOI: http://dx.10.1101/2021.08.04.455025
• 发表时间: 2021
• 作者: Geisler K