Integration of strain engineering with bioprocess engineering strategies to enhance biobased production of value-added chemicals

将菌株工程与生物过程工程策略相结合，以增强增值化学品的生物基生产

• 批准号: RGPIN-2019-04611
• 负责人: Chou, Perry
• 金额: $ 2.4万
• 依托单位: University of Waterloo
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=737739
• 关键词: Integration; strain; engineering; bioprocess; strategies

To date, most chemical products are still made via chemical processing with fossil fuels as raw materials, not only depleting fossil fuels but also generating significant amounts of greenhouse gases (GHGs). Hence, replacing chemical production with modern biobased production potentially offers a true solution to both GHG mitigation and sustainable manufacturing, realizing "low carbon" economy. The proposed research will target biotechnological development of novel engineered bacterial Escherichia coli strains and their associated bioprocessing strategies for clean, effective, economical, renewable, and sustainable production of value-added chemicals. Recently, we successfully demonstrated heterologous production of non-native 1-propanol and propionate in engineered E. coli through activation of a natural sleeping beauty mutase (Sbm) pathway. An important implication associated with our demonstration is that activation of the Sbm pathway introduces a C3-fermentative pathway with the formation of a non-native metabolite intermediate propionyl-CoA in E. coli. The presence of propionyl-CoA opens a wide avenue for novel bacterial synthesis of a selection of non-native high-value bioproducts from unrelated carbon sources using E. coli as a cell factory. In this proposed research, two valuable bioproducts derived from propionyl-CoA, i.e. 3-hydroxyvalerate (3-HV) and valerate, will be used for technological demonstration. First, we will apply various state-of-the-art microbial biotechnologies, including synthetic biology, genetic/genomic engineering, protein engineering, and metabolic engineering to further engineer these propanologenic (propanol-producing) E. coli strains for heterologous production of 3-HV and valerate. Additionally, we will explore these engineered bacterial strains for effective biotransformation using cheap feedstocks, including glycerol, lignocelluloses, and volatile fatty acids. Finally, we will characterize and optimize operating conditions and bioprocessing strategies for large-scale production and evaluate the economic feasibility of developed bioprocesses. It is anticipated that the success of the proposed research in developing effective biobased production platforms will mediate scientific and engineering advances in biomanufacturing, enhance economic feasibility of bioprocessing, and alleviate major environmental impacts caused by traditional chemical processing. The engineered bacterial strains and developed bioprocesses can be readily transferred to our Canadian industrial partners for commercialization, boosting Canada's technological leadership in biomanufacturing. In addition, the proposed research is highly multidisciplinary as its technological aspects cover both fundamental biological sciences and practical biochemical engineering applications; and therefore will be ideal for training of highly qualified personnel (HQP) meeting the challenges of the next generation of industrial biotechnology.

迄今为止，大多数化工产品仍然以化石燃料为原料通过化学加工生产，不仅消耗化石燃料，而且产生大量的温室气体（GHG）。因此，用现代生物基生产取代化学生产可能为温室气体减排和可持续制造提供真正的解决方案，从而实现“低碳”经济。拟议的研究将针对新型工程细菌大肠杆菌菌株的生物技术开发及其相关的生物加工策略，以清洁、有效、经济、可再生和可持续地生产增值化学品。最近，我们成功地证明了通过激活天然睡美人变位酶（Sbm）途径，在工程大肠杆菌中异源生产非天然 1-丙醇和丙酸。与我们的演示相关的一个重要含义是，Sbm 途径的激活引入了 C3 发酵途径，并在大肠杆菌中形成非天然代谢物中间体丙酰辅酶 A。丙酰辅酶A的存在为使用大肠杆菌作为细胞工厂从不相关的碳源中新型细菌合成一系列非天然高价值生物产品开辟了广阔的途径。在这项拟议的研究中，两种有价值的源自丙酰辅酶A的生物产品，即3-羟基戊酸(3-HV)和戊酸，将用于技术示范。首先，我们将应用各种最先进的微生物生物技术，包括合成生物学、遗传/基因组工程、蛋白质工程和代谢工程，进一步改造这些产丙醇（生产丙醇）的大肠杆菌菌株，以异源生产3-HV和戊酸。此外，我们将探索这些工程菌株，以使用廉价的原料（包括甘油、木质纤维素和挥发性脂肪酸）进行有效的生物转化。最后，我们将描述和优化大规模生产的操作条件和生物工艺策略，并评估所开发的生物工艺的经济可行性。预计在开发有效的生物基生产平台方面所提出的研究的成功将促进生物制造的科学和工程进步，提高生物加工的经济可行性，并减轻传统化学加工造成的重大环境影响。工程菌株和开发的生物工艺可以很容易地转移给我们的加拿大工业合作伙伴进行商业化，从而提升加拿大在生物制造方面的技术领先地位。此外，拟议的研究是高度多学科的，因为其技术方面涵盖基础生物科学和实际生化工程应用；因此，它将是培训高素质人才（HQP）应对下一代工业生物技术挑战的理想选择。