Improved enzymes with new functions

Directed evolution: the most revolutionary protein engineering method known to date

We use directed evolution to customized enzymes for different purposes and applications. In this procedure, we first create genetic diversity by random mutagenesis and/or the recombination of parental genes.

Once the mutant libraries are expressed in a suitable host organism, we can screen them in high-throughput platforms to search for specific biochemical traits. These include activity under extreme pH or temperatures, changes in substrate preferences, tolerance in the presence of strong inhibitors…

In this way, we can strictly control selective pressure until the desired properties are attained.

Increased yield and efficiency of industrial processes

Our technological platform EVOSHUFFLER takes biocatalysts to the next level

With an ensemble of top-notch technologies in directed evolution based on the Saccharomyces cerevisiae machinery, we harness a variety of methods aimed at designing customized biocatalysts.

This unique approach offers huge versatility and ad-hoc solutions to different sectors, including the pharmaceutical, chemical, energy and environmental industries.


Get to know the latest work from our team

Engineering a Highly Thermostable High-Redox Potential Lacasse.

Mateljak I, and Alcalde M (2021)

ACS Sustainable Chem. Eng. 2021, 9, 29, 9632–9637

Functional Expression of Two Unusual Acidic Peroxygenases from Candolleomyces aberdarensis in Yeasts by Adopting Evolved Secretion Mutations.

Gómez de Santos, P; Hoang, MD; Kiebist, J; Kellner, H; Ullrich, R; Scheibner, K; Hofrichter, M; Liers, C and Alcalde, M (2021)

Applied and Environmental Microbiology 87 (19): e00878-21

Evolved peroxygenase-aryl alcohol oxidase fusions for self-sufficient oxyfunctionalization reactions.

Gómez-Fernández, B.J., Risso, V.A., Rueda, A., Sánchez-Ruiz, J.M. and Alcalde, M.(2020).

Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases.

Gómez-Fernández, B.J., Risso, V.A., Rueda, A., Sánchez-Ruiz, J.M. and Alcalde, M.(2020).

Applied and Environmental Microbiology 86 (14): e00778-20. JOURNAL COVER.

In vivo site-directed recombination (SDR): An efficient tool to reveal beneficial epistasis. In: Methods in Enzymology 643.

Viña-Gonzalez, J. And Alcalde, M. (2020).

Tawfik, D. Ed. Academic Press Elsevier Inc, Cambridge, MA, EEUU. Pages 1-12. ISBN 978-0-12-821149-6.

Laccase engineering by directed and computational evolution. In: Laccase in bioremediation of pollutants and xenobiotics.

Mateljak, I., Gomez-Fernandez, B. and Alcalde, M. (2020).

Bioremediation and Waste valorization. Schlosser, D., Ed. Springer Nature Switzerland. Pages: 191-206. ISBN: 978-3-030-47906-0

Recent developments in the use of peroxygenases – exploring their high potential in selective oxyfunctionalisations.

Hobisch, M., Holtmann, D., Gomez de Santos, P., Alcalde, M., Hollmann, F. and Kara, S. (2020).

Biotechnology Advances. In press.

Benchmarking of laboratory evolved unspecific peroxygenases for the synthesis of human drug metabolites.

Gomez de Santos, P., Cervantes, F.V., Tieves, F., Plou, F.J., Hollmann, F. and Alcalde, M. (2019).

Research projects

We are involved in publicly funded projects

R&D Projects

2019 – 2022


Synthetic chemistry using fusion chimeras engineered by directed and computational evolution.

2020 – 2023


Apply lignases to resolve end-of-life issues of thermoset composite plastics.

2020 – 2023

Madrid + Circular

Hub de Innovación de Economía Circular de la Comunidad de Madrid.



2020 – 2021

RIS 3 Línea 1.

2020 – 2023

PTQ 2020 – 2023

2021 – 2022

EvoEnzyme. Enzimas Evolucionadas: Soluciones a medida para el sector farmacéutico, de plásticos, químico, energético y medioambiental.

2021 – 2024

PTQ 2021 – 2024

Acceleration programs

Technological platforms