INOFEA AG is a Swiss biotech company founded in 2014 in Basel by a group of entrepreneurs and scientists as a spin-off from the School of Life Sciences, University of Applied Sciences and Arts, Northwestern Switzerland (FHNW). Their high-quality laboratory is dedicated to develop and manufacture immobilized and shielded enzymes with benefits, other enzyme-technologies do not have.
For a more detailed introduction to INOFEA and their efficient enzyme solutions, click here to go to the About INOFEA page.
Prepared below are a series of frequently asked questions about enzymes and the range from INOFEA, take a look through and should you have any more questions or want to find out more, reach out to the team at Brenntag below.
Biocatalysis is a process in which a biological material is used to speed up, or catalyse, a chemical reaction. The biological materials used during biocatalysis are enzymes, protein molecules carrying a vital energy factor needed for reactions that occur in living organisms. Although enzymes have a natural origin, they are often engineered to improve their performance.
Classic metallic and organometallic catalysts deployed in chemical synthesis are often cheap and usually very robust. On the other hand, they are also limited in the specificity and complexity of the reaction they promote, and most of them are highly polluting and complex to dispose.
Enzymes offer incredible features in terms of specificity and specialization of the reaction they catalyse, and they are extremely friendly from an environmental point of view. But they are also fragile and costly to produce. A multitude of factors like acidity, temperature, or the presence of chaotropic agents can affect enzyme structure and their properties. These are the limits that enzzen® technology overcomes.
One of the major challenges in biocatalysis is to transform the physiological catalysts into process catalysts, able to perform an enzymatic reaction under the tough conditions of an industrial process. Enzyme stabilization and protection is INOFEA’s core business. An immobilized enzyme includes the attachment to a solid particle and our shielding increases the stability of enzymes to changes in temperature and pH, in the presence of organic solvents, making them fit for use under typical process conditions.
Any enzyme can be suitable, although we may experience different requirements to customize the immobilization and shielding conditions for each. Examples: for large substrates we recommend a partial layer, for hydrophilic substrates a hydrophilic layer, to enable that the enzymes meet the active site of the enzyme.
The technology is independent from the nature of the beads. We bind the enzyme covalently through a kind of linker to the surface of the carrier material. Other binding strategies are also possible, like through a tag (a kind of tail at one side of the enzyme molecule). The same “anchors” allowing the immobilization of the enzymes are used to secure the enzzen® shield. We are familiar with carries of different nature – mainly silica and polymers – and we keep gaining experience on further options.
Are any residues from the production of enzzen®-products in the final product? After each manufacturing step we do washing steps to ensure that the final immobilized and shielded enzyme does not contain remaining residues.
REACH is a regulation of the European Union, adopted to improve the protection of human health and the environment from the risks that can be posed by chemicals. Manufacturers and importers of substances at one metric ton per year or more must collect information on the properties of those substances and communicate this information to ECHA (European Chemical Agency) through a registration dossier. As enzymes are deployed in insignificant amounts– and we provide customized solutions counting as an individual substance each – we are below the amount requiring such registration.
Through a dedicated proof of concept developed by INOFEA’s R&D. We take the enzyme the customer is interested in, immobilize and shield it and measure various endpoints, such as immobilization yield and enzyme activity after immobilization and shielding. Additionally. INOFEA offers stability and recyclability tests.
It is difficult and potentially misleading to give a general average timing as the needs of our customers and the work to be done can differ. The shortest time to deliver a tangible proof of concept was 2 weeks and the longest so far 9 months while most around 4 months.
As the amount of work can differ significantly it is also the case for the cost, as a natural consequence. Also, in consideration of that we propose modular working packages giving our customer the option to consider if and how to proceed after we get an experience about each specific enzyme/process and we can have a rough estimation of the challenge in its whole.
Of course, we have well established relationships with validated partner CMOs allowing us a full scale up for each enzzen® solution we develop.
Yes, once the enzzen® technology is optimized and validated for the specific needs of our customers we can both offer to use the technology through a license agreement, or we can supply the turnkey enzzen® enzyme in industrial amount.
Enzymes used as biocatalyst (processing aid) in a production do not need to be manufactured under GMP as they are not present in the final product. It is indeed important to avoid enzyme-leakage, which is one of the advantages of enzzen® as it minimizes this risk.
Although we can mention to some extent the fields in which our technology is currently deployed, we are bounded by very strict non- disclosure conditions, in particular – but not only – for pharmaceutical manufacturing applications.
It is suitable and already deployed by our customers in both fields.
As any immobilized enzyme the mechanical resistance in a stirring reactor depends mainly by the nature and the features of the carriers. We carefully choose the beads material to match the requirements of the final application.
We carefully select the right carriers and the size of the beads to ensure the necessary resistance and to optimize the flow so that the pressure does not exceed some limits.
In general the purest the better, but it is not only about quantitative purity, also the nature of impurities matters. In example, as we immobilize enzymes – those are proteins – any other protein can compete in the immobilization subtracting “space” on the beads. Other substances can impact the immobilization, the shielding, and the performances of the enzzen® enzymes. This is an aspect taken in consideration by our R&D team from the early stages of a proof of concept.
The organosilane monomers forming the enzzen® shields can carry a wide range of functionalizations, allowing us to tune significantly the physical and chemical features of the shield surface, i.e., gaining hydrophobicity to ease the interaction between the enzyme’s active site and the substrate.
The implementation and application of our technology has some fix costs which can be easily absorbed, even negligible, by the benefit in term of recyclability and efficacy of valuable enzymes.
We have validated suppliers of enzymes, but we can also work on customer’s own enzymes, managing the customized application of enzzen® and in case providing scale-up options even for the enzyme itself.
Both immobilization and shielding can significantly enhance the activity of an enzyme, but they also limit its freedom, i.e., in terms of orientation and accessibility of the active site. It is important to study in depth each enzyme and its interaction with the beads and the shield to ensure the active site is properly exposed and the substrate access it. Even the substrate may require a very specific tune of the shield to provide the best affinity and accessibility of the active site.
Our experience and the knowledge of both the enzyme and its substrate are a good starting point, but it is always essential to undergo a well-planned series of experiments to validate any theoretical hypothesis. The shield is not a solid cage but more a soft cocoon and allows substrates to migrate to the active side through a kind of tunnel or pores.
Our technology is not an alternative to enzymes immobilization, but actually an improvement of it. We do immobilize the enzymes covalently following the state-of-the-art procedures well established in the industry but optimizing and integrating them in the perspective of the subsequent shielding step that makes our proposition unique.
This depends on many factors, such as the enzymes nature and the conditions of the reaction. What is quite constant is the significant improvement at same conditions compared with the soluble enzyme and the immobilized – but not shielded – enzyme.
Enzzen® strongly enhances the resistance of the enzymes to industrial conditions but can’t grant enzymes to be deployed for any amount of time in any harsh environment. Also, we encountered niche enzymes not suitable for the shielding (i.e., dimeric enzymes). Nevertheless, these are negligible exceptions and we experienced successful deployment in most cases.
Contact the team at Brenntag who will be more than happy to support and answer any questions you may have.
Catarina Gil Forte
Head of Inudstry Market Management, Synthesis & Biopharma
Phone: +31 12 192 48816
Email: catarina.gilforte@brenntag.com
