In vivo directed enzyme evolution in nanoliter reactors with antimetabolite selection

Femmer, Christian; Bechtold, Matthias; Held, Martin; Panke, Sven

doi:10.1016/j.ymben.2020.01.003

Cited by 15 publications

(14 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Moving away from model enrichments and display of platform capabilities, many research groups are now working on improving enzymes with industrial or medical relevance. [ 65 , 74 , 76 , 150 , 152 , 153 ] Besides the evolution of natural enzymes, the toolkit of directed evolution is expanding to artificial enzymes to introduce non‐natural reactivities urgently needed in the pharmaceutical industry, [152] de novo designed enzymes to understand and reengineer enzyme active sites, and, more recently, machine‐learning‐assisted directed evolution. [154] Furthermore, as fluorogenic substrates cannot always be synthesized, novel strategies for fast, but non‐fluorescence‐based detection will be critical for future developments.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Droplet Microfluidics and Directed Evolution of Enzymes: An Intertwined Journey

et al. 2021

View full text Add to dashboard Cite

show abstract

Section: Discussionmentioning

confidence: 99%

“…Starting from a library with 1.2×10 7 variants, they observed an up to 2.7‐fold k cat / K M improvement over wild type after three rounds of directed evolution. [65] …”

Section: Single Emulsionsmentioning

confidence: 99%

Droplet Microfluidics and Directed Evolution of Enzymes: An Intertwined Journey

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Moving away from model enrichments and display of platform capabilities,many research groups are now working on improving enzymes with industrial or medical relevance. [65,74,76,150,152,153] Besides the evolution of natural enzymes,t he toolkit of directed evolution is expanding to artificial enzymes to introduce non-natural reactivities urgently needed in the pharmaceutical industry, [152] de novo designed enzymes to understand and reengineer enzyme active sites,a nd, more recently,m achine-learning-assisted directed evolution. [154] Furthermore,asfluorogenic substrates cannot always be synthesized, novel strategies for fast, but non-fluorescence-based detection will be critical for future developments.…”

Section: Discussionmentioning

confidence: 99%

“…Starting from al ibrary with 1.2 10 7 variants,t hey observed an up to 2.7-fold k cat /K M improvement over wild type after three rounds of directed evolution. [65] 2.2. Technology Advances II:Microfluidics-Based Droplet Formation and Reduction of Cross Talk between Droplets…”

Section: Methodsmentioning

confidence: 99%

Droplet Microfluidics and Directed Evolution of Enzymes: An Intertwined Journey

et al. 2021

View full text Add to dashboard Cite

Evolution is essential to the generation of complexity and ultimately life. It relies on the propagation of the properties, traits, and characteristics that allow an organism to survive in a challenging environment. It is evolution that shaped our world over about four billion years by slow and iterative adaptation. While natural evolution based on selection is slow and gradual, directed evolution allows the fast and streamlined optimization of a phenotype under selective conditions. The potential of directed evolution for the discovery and optimization of enzymes is mostly limited by the throughput of the tools and methods available for screening. Over the past twenty years, versatile tools based on droplet microfluidics have been developed to address the need for higher throughput. In this Review, we provide a chronological overview of the intertwined development of microfluidics droplet‐based compartmentalization methods and in vivo directed evolution of enzymes.

show abstract

“…The group of Sven Panke (ETHZ Basel) recently reported a method for in vivo directed enzyme evolution in nanoliter reactions with anti-metabolite selection [15]. The researchers were interested in producing sulphostin, a dipeptidyl peptidase IV inhibitor and potential type 2 diabetes drug from racemic ornithine.…”

Section: Screeningmentioning

confidence: 99%

Biocatalysis in the Swiss Manufacturing Environment

et al. 2020

View full text Add to dashboard Cite

Biocatalysis has undergone a remarkable transition in the last two decades, from being considered a niche technology to playing a much more relevant role in organic synthesis today. Advances in molecular biology and bioinformatics, and the decreasing costs for gene synthesis and sequencing contribute to the growing success of engineered biocatalysts in industrial applications [1]. However, the incorporation of biocatalytic process steps in new or established manufacturing routes is not always straightforward. To realize the full synthetic potential of biocatalysis for the sustainable manufacture of chemical building blocks, it is therefore important to regularly analyze the success factors and existing hurdles for the implementation of enzymes in large scale small molecule synthesis. Building on our previous analysis of biocatalysis in the Swiss manufacturing environment [2], we present a follow-up study on how the industrial biocatalysis situation in Switzerland has evolved in the last four years. Considering the current industrial landscape, we record recent advances in biocatalysis in Switzerland as well as give suggestions where enzymatic transformations may be valuably employed to address some of the societal challenges we face today, particularly in the context of the current Coronavirus disease 2019 (COVID-19) pandemic.

show abstract

In vivo directed enzyme evolution in nanoliter reactors with antimetabolite selection

Cited by 15 publications

References 48 publications

Droplet Microfluidics and Directed Evolution of Enzymes: An Intertwined Journey

Droplet Microfluidics and Directed Evolution of Enzymes: An Intertwined Journey

Droplet Microfluidics and Directed Evolution of Enzymes: An Intertwined Journey

Biocatalysis in the Swiss Manufacturing Environment

Contact Info

Product

Resources

About