Sustainable Production of Marine Cyanobacterial Products (Sus-C-Products): discovery, scale up and maintenance

越来越多的人认识到蓝藻的生物技术潜力，作为新化合物的来源和绿色工厂的工业原料的生物生产广泛的行业，从生物塑料，制药到高价值的化合物，如颜料。与传统的微生物发现和生产平台不同，蓝藻依赖于光，提供了新的生产方法，也是一种未开发的生物发现资源。分子生物学和光子学的进步使蓝藻更加容易处理。结合特定波长的光，精心控制的培养条件，再加上分子发现技术，开辟了一个大大扩大的微生物发现管道的前景，这可能用于解决紧迫的问题，如发现用于对抗耐药感染和低碳制造的新抗生素。从发现到产品的转变还需要快速轻松地扩大生产规模的能力，并确保在工业规模的培养中保持感兴趣的化学活性。i)实现规模化和i)培养物活性丧失的问题是工业生物技术的两个主要问题领域。我们建议开发一个蓝藻管道的发现和工业利益的新化合物的生产：耦合发现与生产系统的规模好。从微生物和植物中分离的化合物为发现新的化合物和酶提供了一个无与伦比的起点。在过去的30年里，超过70%进入临床试验的抗生素都是基于这类化合物。海洋蓝藻代表了一种特别丰富且大部分未开发的天然产物来源，例如产生一系列有价值的生物活性物质，如多拉司他汀、牙买加酰胺、阿塔那霉素。蓝藻也编码组装具有商业吸引力的化合物，如非蛋白质氨基酸（如聚甲基亮氨酸、聚苯缬氨酸），目前采用的合成方法非常昂贵。其他罕见的基序，如末端炔（具有作为化学生物学工具的巨大潜力）也被编码。我们将使用最先进的基因组学和代谢组学相结合，从蓝藻中挖掘和发现化合物，并介导其可持续和绿色发酵生产。生物合成基因簇将被选择，因为它们有能力介导化学上前所未有的转化。不寻常的酶活性将在其相关的生物合成集群中进行探索，以及作为生物转化的工具进行切除和开发我们提出了一种最先进的、以基因组为主导的方法，而不是使用传统的磨砺和发现方法：1。结合基因组学、代谢和生物活性分析鉴定生物合成基因簇（bgc）。这种基因编码的酶具有生物合成结构新颖的天然产物的潜力。医学上有用的活动。在野生型和异源宿主（专门为每个系统量身定制）中使用异源表达和启动子重构的适当组合来解锁生产。不寻常的酶和新的天然产物。天然产物将被分离纯化，并使用结合详细的质谱分析、基因组分析和1和2DNMR实验的尖端方法进行结构表征（这将为结构表征研究员提供培训）。从重构的WT系统和异源宿主中持续发酵生产化合物是关键。大规模培养对菌株稳定性的影响将使用Xanthella的一系列培养系统进行评估，这些系统允许培养物在非常受控的条件下生长，规模从1l到600L的大规模pbr生长。

There is increasing realization of the biotechnological potential of cyanobacteria as sources of new compounds and as green factories for the bioproduction of industrial feedstocks for a wide range of industries, from bioplastics, pharmaceuticals to high value compounds such as pigments. Unlike conventional microbial discovery and production platforms, cyanobacteria are light dependent, offering novel production methods as well as being an under-explored biodiscovery resource. Advances in molecular biology and in photonics render cyanobacteria much more tractable. Using combinations of specific wavelengths of light, carefully controlled culture conditions coupled with molecular discovery techniques opens up the prospect of a much-enlarged microbial discovery pipeline which may be of use to address pressing problems such as discovery of new antibiotics for use against drug-resistant infections and low carbon manufacturing. To move from discovery to product also requires the ability to scale up production quickly and easily and to ensure that the chemical activity of interest is maintained in cultures at industrial scale. Problems in i) achieving scale up andii) loss of activity in cultures are two major industrial biotech problem areas. We propose to develop a cyanobacterial pipeline for the discovery and production of novel compounds of industrial interest: coupling discovery with production systems that scale well. Compounds isolated from microorganisms and plants provide an unparalleled starting point for new compound and enzyme discovery. Over the past 3 decades more than 70% of antibiotics entering clinical trials have been based on such compounds. Marine cyanobacteria represent a particularly rich and largely untapped source of natural products, for example producing series of valuable bioactives such as dolastatin, jamaicamide, atanapeptin. Cyanobacteria also encode the assembly commercially attractive compounds such as non-proteinogenic amino acids (eg dolamethylleucine, dolaphenvaline) for which current synthetic approaches employed are very costly. Other rare motifs, such as terminal alkynes (with great potential as tools for chemical biology) are also encoded. We will use state-of- the-art combined genomics and metabolomics to mine and discover compounds from cyanobacteria, and mediate their sustainable and green fermentative production. Biosynthetic gene clusters will be selected for their ability to mediate chemically unprecedented transformations. Unusual enzyme activities will be explored both within their associated biosynthetic clusters as well as being excised and developed as tools for biotransformations.3 Rather than using traditional grind and find methods of discovery, we propose a state of the art, genome led approach: specifically we will:1. combine genomic, metabolic and bioactivity analysis to identify biosynthetic gene clusters (BGCs). that encode enzymes with the potential to biosynthesise structural novel natural products with .medicinally useful activities .2. use appropriate combinations of heterologous expression and promotor refactoring within the Wild Type and heterologous host (specifically tailored to each system) to unlock the production of .unusual enzymes and novel natural products .3. natural products will be isolated purified and structurally characterized using cutting edge approached that combine detailed mass spectrometric analysis, genome analysis and 1 and 2DNMR experiments (this will provide training for the fellow in structural characterization. .4. sustainable, fermentative production of the compounds from both the refactored WT systems and the heterologous host is key. The impact of large scale culturing on strain stability will be assessed using Xanthella's range of culturing systems that allow cultures to be grown under very controlled conditions and at scales ranging from 1l to growth in large scale PBRs at 600L.