Optogenetics for integrated, continuous processes for large-scale chemicals manufacture: Next generation manufacturing through synthetic biology

目前使用发酵方法的制造方法的主要限制是在长时间发酵（例如连续和补料分批）期间不能对生产生物体进行实时和非侵入性的闭环反馈控制。定量监测顶部空间中挥发性产物的产物形成，然后使用该信息调节宿主中的生产途径以调节至所需水平的能力将在使用工程生物学方法的化学品、燃料和材料的制造管道中提供稳定性和可预测性。我们的目标是解决这种对挥发性产品的迫切需求，这些产品在反应堆中积聚可能具有细胞毒性。因此，需要对萜烯浓度进行“即时”实时控制，并通过在线产物捕获（例如，喷射;相分离和相关方法）主动去除产物。我们在Scrutton小组中开发了用于活性产物去除的下游处理方法，但缺少捕获产物水平的实时数据并使用该信息提供用于生产生物体/代谢途径的反馈控制策略的能力。我们计划使用光遗传学方法实现这一点，其中质谱仪的读数用于调节生物体/代谢途径。通过直接控制生物反应器LED来调节生产生物体的光遗传学途径。如果这项技术得以实施，将改变使用工程生物学平台的制造业的游戏规则，为通过实时和非侵入性控制实现稳定和可预测的制造提供新的途径。其目的是设计和实施下一代实时监测和反馈控制化学品制造的生物生产过程，使用光响应遗传元件和酶。其目的是利用Scrutton集团的工程化生产菌株建立通用平台能力，并在SYNBIOCHEM/FutureBRH中进一步快速工程化。

A major limitation of current manufacturing methods using fermentation approaches is the inability to have real-time and non-invasive, closed loop feedback control of the producing organism during long fermentations (e.g. continuous and fed-batch). The ability to monitor quantitatively product formation of volatile products in the headspace and then use this information to regulate production pathways in the host to adjust to desired levels would provide stability and predictability in manufacturing pipelines for chemicals, fuels and materials using engineering biology approaches. We aim to address this urgent need for volatile products whose build up in a reactor can be cytotoxic. There therefore needs to be 'on-the-fly' real-time control of terpene concentration and active removal of the product by on-line product capture (e.g. sparging; phase separation and related methods). We have in the Scrutton group develop downstream processing methods for active product removal but missing is the ability to capture real-time data on product levels and use of this information to offer feedback control strategies for use with the producing organisms/metabolic pathways.We plan to achieve this using optogenetic methods where the read out of a mass spectrometer is used to regulate organisms/pathways by direct control of bioreactor LEDs for optogenetic regulation of the producing organism. If made to work this technology would be a game changer for manufacturing using Engineering Biology platforms, offering new routes to stable and predictable manufacture with real-time and non-invasive control.The aim is to design and implement next-generation real-time monitoring and feedback control of bioproduction processes for chemicals manufacturing, using light-responsive genetic elements and enzymes. The purpose is to build a generic platform capability, using engineered production strains in the Scrutton group and the ability to rapidly engineer further in SYNBIOCHEM/FutureBRH.