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

本项目旨在开发耐热酵母菌马氏克鲁维菌，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。马氏K. marxianus可以在高温、低pH和多种不同的糖上生长，这是经济生物过程的重要特性。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中生产的两类重要化学品的生物生产。该项目还将有助于研究生和本科生的培训，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。总体目标是将K. marxianus建立为生产各种生物基化学品的平台宿主。推进K. marxianus的关键是开发新的合成生物学工具，这些工具可以快速创建具有多个基因组编辑的菌株，并可以准确地控制本地和外源基因的转录。该项目通过(1)开发新的工具和方法来解决这些挑战，用于该酵母物种的快速菌株开发和基因组工程，以及(2)应用这些工具进行代谢工程途径，从而产生具有商业意义的天然（醋酸酯）和非天然（聚酮）产品。苯乙基和醋酸异戊酯是有价值的香料化合物，用作工业溶剂。聚酮具有重要的药物价值，包括抗生素、抗癌药物和降胆固醇药物。这个项目背后的基本原理是，K. marxianus是非无菌（和无菌）生物过程中非常有前途的微生物宿主，可以将一系列不同的糖转化为高价值和工业相关的化学物质和蛋白质。这项研究具有变革性，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. marxianus的天然性状，使可再生原料转化为有价值产品的新生物工艺成为可能。该奖项反映了美国国家科学基金会的法定使命，并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，被认为值得支持。

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.