Key expertise

Biocatalysis

Biocatalysis is the usage of enzymes as catalysts of chemical reactions. Enzymes are proteins which form a structure containing an active site, responsible for their catalytic activity. Thanks to this active site, enzymatic transformations are usually selective and allow enantioselectivity and regioselectivity.

Biocatalysis

Biocatalysis allows to access to greener synthesis pathways

Performed under milder conditions, such as aqueous and room temperature, requiring no or residual metal content. Moreover, enzymes are biodegradable catalysts of the reaction and easy to dispose.

Enzymes involved in biocatalytic processes are not anymore limited to lipases and ketoreductases. Nowadays, sources of enzymes for process chemistry applications have expanded: Seqens offers an access to its catalog of enzymes covering all common biocatalysis mediated chemical transformations such as reduction, oxidation, chiral transformation, hydratation, hydrolysis, esterification.

Enzymes / Biocatalysts

SEQENS creates value in bringing best-in-class expertise & knowledge in biology and chemistry tools to develop innovative and greener competitive bioprocesses aiming at manufacturing pharmaceuticals, personal care, chemicals, and phytochemicals.

We offer our customers

Possibilities to identify biocatalysts among a unique biodiversity of microorganisms, with more than 4500 strains (bacteria, Archaea, Fungi, microalgae…) including over 4300 sequenced (meta)genomes. This collection is encompassing a large diversity of microorganisms from extreme environments (deep hydrothermal springs, salt marshes, volcanic environment, oil field…)
More than 500 well characterized enzymes in collection, available on demand, covering 28 families of enzymes across main classes of enzymes (eg oxido-reductases, hydrolases, transferases, biomass treatment, cofactor recycling)
State-of-the-art proprietary technologies for molecular engineering/assay development and screening
Development and optimization of biocatalysis assays from enzyme selection to the industrial scale-up of greener chemical process

Our methodology

Seqens has developed a robust methodology for the development of biocatalytic processes, in two steps: a rapid proof of concept with selection and identification of the first hits. Then the optimization of enzymes as well as optimization of the biocatalytic process toward scale-up industrial development.

Proof of Concept (3 months)

Feasibility study – Proposal

The goal of the feasibility study is to determine the potential interest of the biocatalysis to perform the targeted chemical reaction. SEQENS team will prepare a proposal, typically within two weeks, based on this evaluation. Confidentiality agreement will be in place for this step.

Diversity exploration- Enzymes selection (50 new enzymes)

SEQENS will select 50 new enzymes specifically for the target and will add several more from its own catalogue.

Enzymes expression – Analytical methods (transfer or development)

Enzymes selected in the previous step will be expressed using a microbial host system (bacteria or yeast). The analytical method will be transferred, if provided by the customer, or developed in order to prepare the screening assay.

Screening assay development – Miniaturization

A screening assay will be developed based on the analytical method and adapted to miniaturize the reaction in order to test a large number of enzymes and conditions.

Hit identification

Based on the results of the screening, the best enzymes and conditions (the “hits”) will be used as a starting point for the development of the biocatalytic process.

Optimization and scale-up

Challenge of reaction conditions – Selection of the best candidate (1.5 month)

Conditions of screening assay previously developed will be used in order to refine the conditions to be used. Primary and secondary physicochemical parameters will be challenged for optimal ratio enzyme/substrate/co-factors.

Enzyme evolution – Directed evolution – Computational modeling – (6 months for IP generation)

When a non-natural chemical reaction is targeted, sometimes wild type enzymes have to be improved in order to reach performances compatible with industrialization. This is performed using enzyme engineering, in order to adapt the enzyme structure to its targeted non-natural substrate or conditions (temperature or salt resistance) via modifications of its amino acid sequence. Different methods can be used, such as semi-rational methods which allow to combine in silico modelling, in order to predict the amino acid positions or area to be mutated, with the screening of a large number of enzyme variants. Performing enzyme engineering allows to increase the performances of the enzyme and to generate intellectual property rights on the new variants generated.

Optimization of the reaction conditions (DOE) (2 months)

If needed, the best reaction conditions can be defined using a Design of Experiments approach.

Scale-up of biocatalytic reaction

The biocatalytic reaction will be scaled up, as well as the fermentation process for the production of the enzyme for tailor-made enzymes (more details in the “Production of the biocatalysts: fermentation scale-up” section).

Biocatalysis reaction at pilot scale

The biocatalytic process will then be transferred to the Seqens’Lab for further upscaling towards industrialization.

In silico protein analysis for protein engineering

To optimize enzymes for biocatalysis, it can be important to understand how their conformational dynamics affect their catalytic activity. Conformational dynamics refer to the movements of a protein’s atoms and the resulting changes in its shape

These movements can affect the active site of the enzyme, which is the region where the substrate binds, and the chemical reaction takes place. By understanding the conformational dynamics of enzymes, the key amino acid residues can be identified, and targeted modifications can be applied specifically to favor interactions between residues and substrate having a positive impact on the activity of the enzyme.

There are several techniques in computing that have been used for enzyme evolution, each with its advantages and limitations. The choice of the best technique depends on the specific problem being addressed

In silico design of smart libraries

Homology modeling of the proteins
Identification of hotspots for saturation mutagenesis (HotSpot Wizard)
Analysis of multiple-point mutation combined in active site cavity/tunnel (Carver/Carverdock)
Combining docking, phylogenetic analyses & Rosetta design calculations, QM/MM modeling

Provide better knowledge on catalytic cavity and substrate/co-factor and enzyme interactions)
Prediction of distal active site mutations by conformationally driven Shortest Path Map (SPM)-based enzyme evolution approach

Production of the biocatalysts: fermentation scale-up

For the production of tailor-made enzymes / biocatalysts, a fermentation process has to be developed at the required scale. The most adapted microbial host (bacteria or yeast) will be selected depending on the enzyme form (for example, purified enzyme or whole cells) which is targeted

Seqens has internal fermentation capabilities up to 300L scale, which is compatible with some commercial applications, and has a network of external partners for the implementation of fermentation processes at a larger scale (from m3 scale).

1/ Strain selection, media optimization

Lab-scale (5 – 200 ml) Benchtop bioreactors 1L
Design of Experiments (DoE)
Feed stock (Glucose, Glycerol…)
Fermentation parameters optimization
6x 1L Applikon bioreactors

2/ Fementation process development

Benchtop bioreactor 3L & Pilot plant 40L – 300L
2x 3L Applikon bioreactors
2x 40L Applikon bioreactors
300L Global Process Concept (GPC)
Biomass separation by centrifugation
Downstream process: cell disruption, membrane separation,…

3/ Tech Transfer, Scale-up

Production
Process book
Biocatalyst manufacturing: at SEQENS or transfer to external production site
Biocatalyst reaction: Integration within SEQENS plants or to customer site

Découvrez d'autres services R&D

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Bio
disponibilité

Biodisponibilité

40% des médicaments échouent en essais précliniques en raison de problèmes de biodisponibilité. SEQENS dispose d'une solide expertise pour vous aider à résoudre les problèmes de biodisponibilité

Biologie Synthétique

Biologie de synthèse pour des innovations bioinspirées et durables

Société du groupe SEQENS, Alganelle est une entreprise de biotechnologie innovante à la pointe de la biologie synthétique et de l'ingénierie métabolique

Caracté
risation

Caractérisation des APIs

Les capacités de SEQENS pour la caractérisation physique des principes actifs pharmaceutiques comprennent un ensemble complet de technologies avancées

Immuno
conjugués

Immunoconjugués

Le domaine des immunoconjugués est en train d'émerger, avec la possibilité de réaliser deux fonctions avec un seul composé (bifonctionnel), ce qui permet généralement des thérapies ciblées ou des propriétés de biodisponibilité améliorées

Lipides

Dans l'industrie pharmaceutique, les lipides sont des excipients essentiels pour le développement de systèmes d'administration de médicaments et peuvent être utilisés dans la plupart des formes de dosage