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生产方法的一个主要局限性是不能在长时间发酵(例如连续和补料分批)期间对生产有机体进行实时和非侵入性的闭环反馈控制。能够定量监测顶空中挥发性产品的产品形成，然后使用该信息来调节宿主中的生产路径，以调整到所需的水平，这将在使用工程生物学方法制造化学品、燃料和材料的管道中提供稳定性和可预测性。我们的目标是满足这种对挥发性产品的迫切需求，这些产品在反应堆中的积累可能会对细胞产生毒性。因此，需要通过在线产品捕获(例如，喷雾、相分离和相关方法)来实时控制萜烯浓度并主动去除产品。我们在Srutton团队中开发了用于去除活性产物的下游处理方法，但缺少捕获产品水平的实时数据并使用这些信息提供反馈控制策略的能力，以用于生产生物体/代谢途径。我们计划使用光遗传方法实现这一点，其中，通过直接控制生物反应器LED来调节生物/代谢途径，使用质谱仪读数来调节生物/途径。如果成功，这项技术将改变使用工程生物学平台进行制造的游戏规则，提供稳定和可预测的制造的新途径，具有实时和非侵入性控制。目标是设计和实施下一代实时监测和反馈控制，使用光响应性基因元件和酶来制造化学品的生物生产过程。其目的是利用斯克鲁顿集团的工程生产菌株以及在SYNBIOCHEM/FutureBRH中快速设计进一步工程的能力，建立通用平台能力。

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.