Bioinspired alkene cleavage - BioZone

The aim of this project is to develop a biological (enzyme-assisted) method for splitting carbon / carbon double bonds without the use of the dangerous ozone. The so-called ozonolysis is a widely used method for the production of so-called "carbonyl compounds". Such carbon / carbon double bond containing compounds can be starting materials to make such as flavor compounds like vanillin or fragrances like Lilial. Ozonolyis is mainly used on a laboratory scale. The reason for this is the toxicity of ozone and the danger of the intermediate products, which are not safe: Some of them have already caused industrial reactors to explode. We want to find and investigate a safe alternative with enzymes that can split C = C double bonds at room temperature, in water and by means of oxygen. We use a high-throughput detection method that is specifically designed for the detection of aldehydes (carbonyl compounds) in order to examine different protein sequences for their ability to split C = C double bonds. This detection method is highly sensitive, completely independent of structure, easy to use and extremely specific for aldehydes. With this detection method in combination with a special technique, it is possible to examine up to a million protein sequences in a day. It is thus possible to examine known enzymes for their substrate diversity and new enzymes for their activity in a very short time, as quantitatively as possible. After suitable enzymes have been found, we will use them for the production of potential new active pharmaceutical ingredients or flavorings.

The BioZone project focused on developing an environmentally friendly alternative for producing aldehydes from unsaturated hydrocarbons. These "aldehydes" are important chemical compounds used in the fragrance and flavouring industry. One example is vanilla, which is a must-have in every kitchen, especially during the Christmas season. Another example is cinnamaldehyde, which is also a commonly used spice with a characteristic smell. The chemical production of these compounds is expensive, often involves highly toxic or dangerous reagents, and is not very efficient. That is why we have developed a method in this project that is based on enzymes, known as "biocatalysts". To this end, we produced new enzymes, tested them on a laboratory scale and further developed the best candidates for real industrial application. In addition, we attempted to gain a deeper insight into the function of this biocatalyst. To do this, we crystallised the enzyme and thus elucidated its exact structure. With this knowledge, we further improved the enzyme and finally used it in combination with other biocatalysts to produce vanilla from the renewable raw materials lignin or ferulic acid. We have thus succeeded in establishing a more sustainable method of producing vanilla. The next steps are now to transfer this method to an industrial context.