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: 1803677
• 负责人: Nancy Da Silva
• 金额: $ 31.5万
• 依托单位: University of California-Irvine
• 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 2023-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1803677&HistoricalAwards=false
• 关键词: Collaborative; Research; SusChEM; Engineering; thermotolerant

This project seeks to develop the yeast Kluyveromyces marxianus as a platform microorganism for industrial biomanufacturing. 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 in high acidity, and on a wide range of different sugars. These are 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.

该项目旨在开发马克斯克鲁维酵母作为工业生物制造的平台微生物。美国工业生物技术领域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。马克斯克鲁维酵母可以在高温、高酸度和多种不同的糖上生长。 这些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，并应用这些工具来工程化传统上由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学院学生开发新的教育推广项目。总体目标是将 K. marxianus 建立为生产多种生物基化学品。发展马克斯克鲁维酵母的关键是开发新的合成生物学工具，该工具可以快速创建具有多个基因组编辑的菌株，并且可以准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) 开发新的支持工具和方法，用于该酵母物种的快速菌株开发和基因组工程；(2) 应用代谢工程工具，对产生具有商业意义的天然（醋酸酯）和非酯的途径进行代谢工程。 -天然（聚酮化合物）产品。苯乙基和乙酸异戊酯是有价值的香料/香料化合物，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、抗癌和降胆固醇药物。该项目背后的基本原理是，马克斯克鲁维酵母是一种非常有前途的非无菌（和无菌）生物过程微生物宿主，可以将一系列不同的糖转化为高价值和工业相关的化学品和蛋白质。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 的天然特性，从而实现将可再生原料转化为有价值产品的新生物过程。该奖项反映了 NSF 的法定使命，并被视为值得通过使用基金会的智力优点和更广泛的影响审查标准进行评估来支持。

项目成果

Synthesis of polyketides from low cost substrates by the thermotolerant yeast Kluyveromyces marxianus

耐热酵母马克斯克鲁维酵母以低成本底物合成聚酮化合物

• DOI: 10.1002/bit.26976
• 发表时间: 2019-07-01
• 期刊: Biotechnology and Bioengineering
• 影响因子: 3.8
• 作者: Tami L. McTaggart;D. Bever;Shane Bassett;N. D. Da Silva
• 通讯作者: N. D. Da Silva

Enhanced production of acetyl-CoA-based products via peroxisomal surface display in Saccharomyces cerevisiae

通过酿酒酵母中的过氧化物酶体表面展示提高基于乙酰辅酶A的产品的产量

• DOI: 10.1073/pnas.2214941119
• 发表时间: 2022-11-21
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Hannah C Yocum;Shane Bassett;N. D. Da Silva
• 通讯作者: N. D. Da Silva

RNA polymerase II-driven CRISPR-Cas9 system for efficient non-growth-biased metabolic engineering of Kluyveromyces marxianus

RNA聚合酶II驱动的CRISPR-Cas9系统用于马克斯克鲁维酵母的高效非生长偏向代谢工程

• DOI: 10.1016/j.mec.2022.e00208
• 发表时间: 2022-12
• 期刊: Metabolic Engineering Communications
• 影响因子: 5.2
• 作者: Bever D;Wheeldon I;Da Silva N
• 通讯作者: Da Silva N

Assembly of Metabolons in Yeast Using Cas6-Mediated RNA Scaffolding

使用 Cas6 介导的 RNA 支架组装酵母中的代谢物

• DOI: 10.1021/acssynbio.2c00650
• 发表时间: 2023-04
• 期刊: ACS Synthetic Biology
• 影响因子: 4.7
• 作者: Pham, Anhuy;Bassett, Shane;Chen, Wilfred;Da Silva, Nancy A.
• 通讯作者: Da Silva, Nancy A.

CRISPR‑mediated multigene integration enables Shikimate pathway refactoring for enhanced 2‑phenylethanol biosynthesis in Kluyveromyces marxianus

CRISPR介导的多基因整合能够重构莽草酸途径，从而增强马克斯克鲁维酵母中的2-苯乙醇生物合成

• DOI: 10.1186/s13068-020-01852-3
• 发表时间: 2021-01-06
• 期刊: Biotechnology for biofuels
• 影响因子: 6.3
• 作者: Li M;Lang X;Moran Cabrera M;De Keyser S;Sun X;Da Silva N;Wheeldon I
• 通讯作者: Wheeldon I