Collaborative Research: SusChEM: Engineering the thermotolerant yeast Kluyveromyces marxianus for the synthesis of biobased chemicals

合作研究：SusChEM：改造耐热酵母马克斯克鲁维酵母用于合成生物基化学品

• 批准号: 1803630
• 负责人: Ian Wheeldon
• 金额: $ 31.53万
• 依托单位: University of California-Riverside
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1803630&HistoricalAwards=false
• 关键词: Collaborative; Research; SusChEM; Engineering; thermotolerant

This project seeks to develop the thermotolerant yeast Kluyveromyces marxianus as a platform microorganism for industrial bioprocessing. A critical area of the US industrial biotechnology sector is the conversion of biomass and other renewable feedstocks to high value and commodity chemicals. K. marxianus can grow at high temperature and low pH, and on a wide range of different sugars, important traits for economic bioprocesses. The project will develop new genetic engineering tools for this yeast and apply these tools to engineer the bio-production of two important classes of chemicals that are traditionally produced from petroleum feedstocks. The project will also contribute to the training of graduate and undergraduate students, as well as the development of new educational outreach programs for high school and community college students from diverse backgrounds.The overall goal is to establish K. marxianus as a platform host for the production of a broad range of biobased chemicals. Critical to advancing K. marxianus is the development of new synthetic biology tools that can rapidly create strains with multiple genome edits and that can accurately control transcription of native and heterologous genes. This project addresses these challenges by (1) developing new enabling tools and methods for rapid strain development and genome engineering in this yeast species, and (2) applying the tools for metabolic engineering of pathways leading to commercially significant native (acetate esters) and non-native (polyketide) products. Phenylethyl and isoamyl acetate are valuable fragrance/flavor compounds and are used as industrial solvents. Polyketides have significant value as pharmaceuticals, including as antibiotic, anticancer, and cholesterol-lowering drugs. The rationale behind this project is that K. marxianus is a highly promising microbial host for non-aseptic (and aseptic) bioprocesses that can convert a range of different sugars into high-value and industrially-relevant chemicals and proteins. This research is transformational because it develops new synthetic biology tools and metabolic engineering approaches to harness and enhance the native traits of K. marxianus, enabling new bioprocesses for the conversion of renewable feedstocks to valuable products.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该项目旨在开发耐热酵母Kluyveromyces marxianus作为工业生物加工的平台微生物。美国工业生物技术部门的一个关键领域是将生物质和其他可再生原料转化为高价值和商品化学品。K. marxianus可以在高温和低pH值下生长，并在各种不同的糖上生长，这是经济生物过程的重要特征。该项目将为这种酵母开发新的基因工程工具，并应用这些工具来设计传统上从石油原料生产的两类重要化学品的生物生产。该项目还将有助于培养研究生和本科生，以及为来自不同背景的高中和社区大学学生开发新的教育推广计划。marxianus作为生产广泛的生物基化学品的平台主机。关键在于提高K。marxianus是新的合成生物学工具的开发，可以快速创建具有多个基因组编辑的菌株，并且可以精确控制天然和异源基因的转录。该项目通过以下方式解决这些挑战：（1）开发新的使能工具和方法，用于该酵母物种的快速菌株开发和基因组工程，以及（2）将工具应用于导致商业上重要的天然（乙酸酯）和非天然（聚酮化合物）产品的途径的代谢工程。乙酸苯乙酯和乙酸异戊酯是有价值的香料/风味化合物，用作工业溶剂。聚酮化合物作为药物具有重要价值，包括作为抗生素、抗癌药和降胆固醇药。这个项目背后的基本原理是K。Marxianus是用于非无菌（和无菌）生物过程的非常有前途的微生物宿主，其可以将一系列不同的糖转化为高价值和工业相关的化学品和蛋白质。这项研究是变革性的，因为它开发了新的合成生物学工具和代谢工程方法来利用和增强K的天然特性。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识价值和更广泛的影响审查标准进行评估来支持。

项目成果

Highly Multiplexed CRISPRi Repression of Respiratory Functions Enhances Mitochondrial Localized Ethyl Acetate Biosynthesis in Kluyveromyces marxianus

• DOI: 10.1021/acssynbio.8b00331
• 发表时间: 2018-11-01
• 期刊: ACS SYNTHETIC BIOLOGY
• 影响因子: 4.7
• 作者: Lobs, Ann-Kathrin;Schwartz, Cory;Wheeldon, Ian
• 通讯作者: Wheeldon, Ian