Optimising strain selection and developing a biorefinery for industrially relevant micro-algae

授予：BB/T50872 X/1 BBRSC本提案的主要目的是通过采用不同的藻类生物精炼方法来提高商业规模藻类生物技术的潜力。候选种属将根据其生化特征和细胞特征、易培养性和商业化潜力进行选择。例如，小球藻属，微拟球藻属，Phaeodactylum sp.和Tisochyrsis sp.都能够合成一系列色素、类胡萝卜素和必需脂肪酸，这使得它们对营养、制药和水产养殖饲料工业具有潜在价值。此外，它们都具有不同的细胞形态，这使它们对下游加工具有不同程度的依赖性。为了从产生的生物质中获得最高价值，将研究各种细胞破碎（冻融、珠磨、超声处理）和产物纯化（过滤、离心、色谱）技术。然后可以在大规模光生物反应器中进行可行性研究，以便更好地概述工业微藻生物技术的商业潜力。另一个症结是缺乏针对正在开发的微藻物种的培养基和条件的定制培养。这可能对所生产的生物质的质量和数量产生重大影响。为了解决这些问题，学生将与学术合作伙伴SAMS（UHI）和工业合作伙伴Xanthella密切合作，因为这两个组织位于同一地点，这将进一步促进有关生产方法/规模和生物精炼的反馈正在进行的方法。具体而言，我们将：项目A。最初，将根据专业知识、适用性/最终用途、可销售性和立法（即GRAS认证）选择候选微藻物种（第0-3个月）。B.一套方法将开发和优化，将在整个学生期间使用，以评估生长（血细胞计数器/光密度/库尔特计数器）和藻类生物质的生化组成（分光光度法/HPLC/GC-MS）。将使用高通量筛选方法（多孔板读数器）确定每种候选种属的最佳生长条件。（0-18个月）。C.将测试培养条件（例如，使用CT室的培养基配方、盐度、光照和温度方案、Algem环境建模PBR和Xanthella的PBR以及选择性育种），以修改细胞特性，从而减轻细胞破坏并提高产品产量（第6-24个月）。d.奖励最有前途的候选物种将在更大规模的培养，使用microPharos和Pandora光生物反应器（PBRs），与生物精炼框架保持一致，这将在Xanthella有限公司完成（第18-36个月）。e.将使用多种技术（例如冻融/珠磨/超声处理）破坏藻类生物质。将使用直接和间接方法（细胞计数/生物化学/显微镜/流式细胞术结合人工辅助机器学习）评估破坏效率。将努力在细胞破坏步骤期间实现选择性释放，以促进进一步的分级分离步骤。为此，由于特定组分位于不同的细胞区室或细胞器上，因此需要了解细胞拓扑结构以设计利用这种区室化的选择性释放策略（第6-36个月）。f.奖励将在逐个物种的基础上探索不同的生物精炼战略。战略将建立在监督团队以前的经验基础上，并将考虑目标产品的利益，可提取性和技术经济分析（第6-36个月）。督导小组：Michele Stanley教授、Dr. Jubeau、Dr. Valeria Montalescot和Dr. Michael Ross

Grant: BB/T50872X/1 BBRSCThe main objective of this proposal is to improve the potential for commercial-scale algal biotechnology by employing different algal biorefinery approaches. Candidate species will be selected based upon their biochemical profile and cellular characteristics, ease of cultivation, and commercialisation potential. For example, Chlorella spp., Nannochloropsis spp., Phaeodactylum sp. and Tisochyrsis sp. are all capable of synthesising an array of pigments, carotenoids and essential fatty acids that make them potentially valuable for nutrition, pharmaceutical and aquaculture feed industries. Furthermore, they all have different cell morphologies which give them varying degrees of recalcitrance to downstream processing. In order to obtain the highest value from the biomass produced, a variety of cell disruption (freeze/thawing, bead milling, sonication) and product purification (filtration, centrifugation, chromatography) techniques will be investigated. Feasibility studies can then be performed in large-scale photobioreactors to give a better overview of the commercial potential of industrial micro-algal biotechnology. Another sticking point is the lack of tailoring culture of media and conditions to the microalgal specie being exploited. This can have a major impact in terms of the quality and quantity of biomass produced. In order to address these issues the student will work closely with both the academic partner, SAMS (UHI), and the industrial partner, Xanthella, since both organisations are on the same site this will further facilitate feedback in relation to the production methods/scale and biorefinery approaches being undertaken. Specifically we will: The project a. Initially candidate micro-algal species will be selected based upon expertise, applicability/end-use, and marketability and legislature (i.e. GRAS certified)(Month 0-3). b. A suite of methods will be developed and optimised that will be used throughout the studentship to assess growth (haemocytometer/optical density/Coulter counter) and the biochemical composition of the algal biomass (spectrophotometry/HPLC/GC-MS). Optimal growing conditions for each candidate species will be determined using a high-throughput screening approach (multi-well plate reader). (Month 0-18). c. Cultivation conditions (e.g. media formulation, salinity, light and temperature regime using CT rooms, Algem environmental modelling PBR's and Xanthella's PBRs, and selective breeding) will be tested to amend cellular properties with a view to ease cell disruption and enhance product yield (Month 6-24). d.Reward The most promising candidate specie(s) will be cultivated at a larger-scale, using the microPharos and Pandora photobioreactors (PBRs), in alignment with the biorefinery framework this will completed at Xanthella Ltd premises (Month 18-36). e. Algal biomass will be disrupted using a variety of techniques (e.g. freeze-thaw/bead milling/sonication). Disruption efficiency will be assessed using both direct and indirect methods (cell counts/biochemistry/microscopy/flow cytometry coupled with human-assisted machine learning). Efforts will be made to achieve selective release during the cell destruction step to facilitate further fractionation steps. For this purpose, as specific components are located on different cell compartments or organelles, the understanding of cellular topology is required to design a selective release strategy that take advantage of this compartmentalisation (Month 6-36). f.Reward Different biorefinery strategies will be explored on a species-by-species basis. Strategies will be founded upon the previous experiences of the supervisory team and will take into account target product(s) of interest, extractability, and techno-economic analysis (Month 6-36). Supervisory Team: Professor Michele Stanley, Dr Sebastien Jubeau, Dr Valeria Montalescot and Dr Michael Ross