H2020Индивидуална стипендия2017–2019

TreasureDrop · Directed Evolution of Enzyme for Applied Biocatalysis at Ultrahigh Throughput in Picoliter Droplets

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2017-05-01 → 2019-04-30
Финансиране от ЕС: 183 455 €
Участници: 1
Схема: MSCA-IF-EF-SE

Линиите свързват координатора с партньорите.

Накратко на български

Ензимите се подобряват чрез бързо筛选 на милиони микрокапки, за да се открият по-ефективни биокатализатори за индустриални цели. Методът позволява много по-бързо откриване на нови или модифицирани протеини, които работят по-добре при конкретни процеси.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

Directed Evolution of Enzyme for Applied Biocatalysis at Ultrahigh Throughput in Picoliter Droplets

Enzymes have established as a new class of catalysts, but remain insufficient in terms of process compatibility, activity or selectivity. Enzyme engineering – i.e. directed evolution or semi-rational protein design – is currently the most promising approaches to improve or alter known enzymes. Further, the field of metagenomics bears high potential to find new scaffolds. In either case, the detection of enzymatic activity via an optical read-out is a simple, fast and versatile tool to find desired biocatalyst. However, a throughput of ≤105samples/day using plate formats is limiting the potential of these assays. Droplet-based single-cell assays boost the chances to succeed by enabling ultra-high throughput (uHTP) screening of ≥107samples/day – if considering that success often correlates with numbers. We aimed at using this technique to showcase a proof-of-principal enzyme engineering campaign targeting an industrially relevant application. In addition, its potential for functional metagenomics was investigated. In conclusion we are convinced that the main objectives of the action were not just reached, but even exceeded based on the results obtained during the last 2 years. This was possible due to an outstanding support from all project participants and access to the newly established robotic facilities at JM. The microfluidic assay technology has been confirmed as a highly promising technique to not only access better, but also new enzymes for industrial applications. The action was recently selected by the Innovation Radar and its exploitation as a service to customers is currently being discussed at JM. Even after the end of the action, JM and the university are in close contact (including me advising students) and are discussing future collaboration opportunities.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

Enzymes have established as a new class of catalysts in the field of modern synthetic chemistry and continue to gain in importance. Directed evolution is currently one of the most promising approaches aiming at enzymes with desired catalytic activities and it's potentially directly correlates with the library size that can be screened. One of the most powerful approaches to overcome these limitations is arguable the recently introduced microfluidic droplet technology; this methodology not only allows to quickly screen millions of clones in a cost effective manner, but is also broadly applicable since fluorometric as well as colorimetric assays can be used. Interestingly, even though numerous publication highlight its potential, an unambiguous evidence of its ability to provide synthetically relevant biocatalysts still needs to be furnished. In addition, access to this technology is currently limited to a few academic research groups and thus, this approach requires further implementation to evolve as an easily manageable lab routine in the near future. This project is designed to unite three competencies: i) the expertise of the Hollfelder Group in regarding micro-engineering and protein engineering in droplets, ii) the empirical knowledge of (bio)chemists at Johnson Matthey in view of economically successful industrial applications of biocatalysts and iii) the strong track record of the experienced researched to successfully solve problems at the biology/chemistry-interface. The objective of the project is to perform a proof-of-principle study by improving a well-known alcohol dehydrogenase for the selective desymmetrization of a meso-diol, thereby giving access to a synthetically sophisticated alcohol. In addition, the final aim is not only to obtain an improved mutant which allows to perform the selected biotransformation efficiently, but also a comparison of varying evolution paths differing in the criteria of hit selection between mutagenesis rounds.

Оригинален текст от CORDIS (на английски).

Участници

JOHNSON MATTHEY PLC · LONDONКоординаторОбединеното кралство