Directed evolution of material- producing microorganisms

• 成果类型: Article
• 署名作者: Laurent, Julie M.; Jain, Ankit; Kan, Anton; Steinacher, Mathias; Casimiro, Nadia Enrriquez; Stavrakis, Stavros; deMello, Andrew J.; Rogers, John
• 署名单位: Swiss Federal Institutes of Technology Domain; ETH Zurich; Swiss Federal Institutes of Technology Domain; ETH Zurich
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-10227
• DOI: 10.1073/pnas.2403585121
• 发表日期: 2024-07-30
• 关键词: bacterial; biosynthesis; mutations; proteins; complex; cells; image

摘要

Nature is home to a variety of microorganisms that create materials under environmentally friendly conditions. While this offers an attractive approach for sustainable manufacturing, the production of materials by native microorganisms is usually slow and synthetic biology tools to engineer faster microorganisms are only available when prior knowledge of genotype-phenotype links is available. Here, we utilize a high- throughput directed evolution platform to enhance the fitness of whole microorganisms under selection pressure and identify genetic pathways to enhance the material production capabilities of native species. Using Komagataeibacter sucrofermentans as a model cellulose- producing microorganism, we show that our droplet- based microfluidic platform enables the directed evolution of these bacteria toward a small number of cellulose overproducers from an initial pool of 40,000 random mutants. Sequencing of the evolved strains reveals an unexpected link between the cellulose- forming ability of the bacteria and a gene encoding a protease complex responsible for protein turnover in the cell. The ability to enhance the fitness of microorganisms toward a specific phenotype and to unravel genotype-phenotype links makes this high- throughput directed evolution platform a promising tool for the development of new strains for the sustainable manufacturing of materials.