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: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=747118
• 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，大多数化学产品仍然是通过化石燃料作为原材料的化学加工制造的，不仅会耗尽化石燃料，而且还产生了大量的温室气体（GHG）。因此，用现代生物生物生产代替化学生产可能为缓解温室气体和可持续制造提供了真正的解决方案，从而实现了“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发展及其相关的生物处理策略，以清洁，有效，经济，可再生和可持续的增值化学品生产。最近，我们通过激活天然的睡美人突变酶（SBM）途径成功地证明了在工程大肠杆菌中的非本地1-丙醇和丙酸酯的异源产生。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，并形成了大肠杆菌中非本地代谢物中间丙酰基-COA。丙酰辅酶A的存在为新的细菌合成开辟了广泛的途径，该途径是使用E. Coli用作细胞工厂的无关碳源中从无关的碳源中选择的非本地高价值生物产品。在这项拟议的研究中，将使用源自丙酰辅酶A的两种有价值的生物产品，即3-羟基效果（3-HV）和valerate，用于技术演示。首先，我们将采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，以进一步设计这些促丙基苯甲酸（丙醇产生）大肠杆菌菌株，用于3-HV和Valreate的异型生产。此外，我们将使用廉价的原料（包括甘油，木质纤维素和挥发性脂肪酸）探索这些工程的细菌菌株，以进行有效的生物转化。最后，我们将表征和优化大规模生产的运营条件和生物处理策略，并评估开发的生物处理的经济可行性。可以预料，拟议的研究在开发有效的生物生产平台方面的成功将调解生物制造的科学和工程进步，增强生物处理的经济可行性，并减轻传统化学处理引起的重大环境影响。可以很容易地将工程的细菌菌株和开发的生物处理转移到我们的加拿大工业合作伙伴中以进行商业化，从而促进加拿大在生物制造方面的技术领导力。此外，拟议的研究是高度多学科的，因为其技术方面涵盖了基本的生物科学和实用的生化工程应用。因此，非常适合培训高素质的人员（HQP）应对下一代工业生物技术的挑战。