NEW — Version 3.0

EVO UPOkit 3.0

Enzymes for the chemical and pharmaceutical sectors

The first screening kit in the market with a selection of evolved fungal unspecific peroxygenases (UPOs). P450 chemistry. H₂O₂ simplicity. Designed to achieve scalable production of chemicals and human drug metabolites.

16

evolved UPO
variants

160

reactions
per kit

H₂O₂

sole
oxidant

2

UPO
families
What is in the kit
16 evolved UPO variants (lyophilized)

Glass vials — short and long-type UPO families from different fungal species

2 reaction buffers

Optimised for UPO activity across different substrates

2 hydrogen peroxide vials

Pre-calibrated for assay-ready use

Positive control substrate

UPO standard substrate for activity validation

Step-by-step protocol

With technical support from our scientists

160 reactions per kit. Custom formats and immobilized enzyme versions available on request.

The UPO advantage

Why UPOs over
P450 monooxygenases?

UPOs emulate the selective oxyfunctionalization chemistry of human P450s — without their complex cofactor requirements. The result: the same powerful chemistry, dramatically simpler to use.

P450 monooxygenases
Requires NADPH cofactor
Expensive, unstable — needs regeneration system.
Membrane-bound enzymes
Difficult to express, purify and scale.
Low stability
Short half-life, sensitive to process conditions.
Complex assay setup
Multiple components, specialised equipment needed.
Evo UPOkit peroxygenases
H₂O₂ as sole oxidant
Simple, cheap, no regeneration system required.
Secreted, soluble enzymes
Easy to produce, purify and handle.
Engineered thermostability
Evolved variants — stable under process conditions.
Assay-ready format
Protocol included — results from day one.
Catalytic repertoire

Selective oxyfunctionalizations
on complex organic compounds.

The UPOs in EVO UPOkit perform an extensive range of selective reactions on pharmacological compounds — emulating the role of human liver P450 monooxygenases.

Hydroxylations

Alkyl and aromatic hydroxylations — introduction of –OH groups with high regioselectivity. Key reaction for human drug metabolite synthesis.

e.g. dextromethorphan → dextrorphan · naproxen · tolbutamide metabolites

O/N-Dealkylations

Ether and ester cleavage (O-dealkylation) and N-dealkylation reactions — critical for phase I drug metabolism studies.

e.g. O-demethylation · N-deethylation · ester cleavage

Epoxidations

Aliphatic and aromatic epoxide formation

S-Oxidations

Sulfide to sulfoxide and sulfone

N-Oxidations

Nitrogen oxidation reactions

Brominations

Selective aromatic bromination

case studies

Published applications.
Real results.

Applications documented in peer-reviewed publications — from human drug metabolite synthesis to late-stage functionalization.

Human drug metabolites · Pharma
In-house · HDM synthesis
Synthesis of authentic human drug metabolites from dextromethorphan, naproxen and tolbutamide

Two engineered UPO variants produced authentic HDMs from three pharmaceutical compounds of interest — demonstrating direct application in drug discovery and metabolite reference standard production

EvoEnzyme in-house study · data available on request

Enantioenrichment · Fine chemistry
Angewandte Chemie 2024
Peroxygenase-Enabled Reductive Kinetic Resolution for the Enantioenrichment of Organoperoxides

UPO variants from our portfolio used for reductive kinetic resolution — demonstrating the versatility of evolved peroxygenases beyond standard oxyfunctionalization

Shen et al. · Angewandte Chemie 63(21), e202401590 · 2024

Bioactive compounds · Food/health
New Biotechnology 2024
Exploiting UPO versatility to transform rutin into more soluble and bioactive products

UPO-mediated transformation of the flavonoid rutin — producing hydroxylated derivatives with improved aqueous solubility and enhanced biological activity

Muñiz-Mouro, Gullón & Eibes · New Biotechnology 83, 197–204 · 2024

Your substrate
Exploratory assay

Have a specific substrate you’d like to test?

Send us your compound — we run the exploratory assay at our facilities and deliver a full report with the best UPO candidates and optimisation recommendations.

From kit to scale

The kit is the first step.

Step 1

Screen with the kit

Test 16 UPO variants on your substrate with the included protocol

You are here
Step 2

Identify your hit

Best variant on your substrate — confirmed by activity data

Screening output
Step 3

Order in bulk

Hit variant from 100 mg to kg — Pichia or yeast expression

Scale-up
Step 4

Evolve further

Custom directed evolution campaign to optimise further

Optional

A Pichia pastoris expression system has been adapted and production protocols established for bioreactor secretion of soluble, very stable, extracellular peroxygenases at larger scale.

Additional services

Beyond the kit.
Full support.

Scale-up production of evolved peroxygenases

Larger quantities of any evolved UPO available on request. Established bioreactor protocols for soluble, extracellular peroxygenase secretion — with downstream handling and secure supply.

Tailored EVO UPOkit

Custom kit formats for specific testing scenarios — adapted for multiple chemical entity screening. Immobilized enzyme versions also in development for flow chemistry applications.

In-house testing and analysis

We run the exploratory assay at our facilities for a specific biotransformation or challenge. After careful analysis, a detailed report identifies the best candidate enzymes and optimisation recommendations.

Peroxygenase customization via Directed Evolution

Once the best candidate is chosen, we offer further development through ad-hoc directed evolution campaigns — including HTP assay design, library creation via EVOSHUFFLER® and scale-up production.

Late-stage oxyfunctionalization services

The EVO UPOkit toolbox is particularly useful in drug discovery pipelines for late-stage functionalization — introducing functional groups and screening chemical libraries to generate new molecular entities.

Not sure where to start?

Our scientists will analyse your specific needs and recommend the best approach — whether it’s the kit, an exploratory assay or a full custom project.

The initial development of the EVO UPOkit was supported by a grant co-funded by the European Regional Development Fund (ERDF/FEDER) under the 2019 call for the development of young innovative technology-based companies (Start-ups) and high innovation-intensity SMEs, within the framework of the regional strategy for Research and Innovation for Smart Specialisation (RIS3), under the ERDF/FEDER operational programme of the Community of Madrid. CAM RIS3 Line 1. Project No. S-2019/L1-274.

Ready to try EVO UPOkit
on your substrate?

Request information, order a kit, or send us your substrate for an exploratory assay — our scientists will get back to you within 24–48 hours.