Industrial applications of immobilized enzymes—A review

The use of immobilized enzymes is now a routine process for the manufacture of many industrial products in the pharmaceutical, chemical and food industry. Some enzymes, such as lipases, are naturally robust and efficient, can be used for the production of many different molecules and have a wide range of industrial applications thanks to their broad selectivity. As an example, lipase from Candida antarctica (CalB) has been used by BASF to produce chiral compounds, such as the herbicide Dimethenamide-P, which was previously made chemically. The use of the immobilized enzyme has provided significant advantages over a chemical process, such as the possibility to use equimolar concentration of substrates, obtain an enantiomeric excess > 99%, use relatively low temperatures (< 60 °C) in organic solvent, obtain a single enantiomer instead of the racemate as in the chemical process and use a column configuration that allows dramatic increases in productivity. This process would not have been possible without the use of an immobilized enzyme, since it runs in organic solvent [1]. Some more specific enzymes, like transaminases, have required protein engineering to become suitable for applications in production of APIs (Active Pharmaceutical Ingredients) in conditions which are extreme for a wild type enzyme. The process developed by Merck for sitagliptin manufacture is a good example of challenging enzyme engineering applied to API manufacture. The previous process of sitagliptin involved hydrogenation of enamine at high pressure using a rhodium-based chiral catalyst. By developing an engineered transaminase, the enzymatic process was able to convert 200 g/l of prositagliptin in the final product, with e.e. >99.5% and using an immobilized enzyme in the presence of DMSO as a cosolvent [2]. For all enzymes, the possibility to be immobilized and used in a heterogeneous form brings important industrial and environmental advantages, such as simplified downstream processing or continuous process operations. Here, we present a series of large-scale applications of immobilized enzymes with benefits for the food, chemical, pharmaceutical, cosmetics and medical device industries, some of which have been scarcely reported on previously. In general, all enzymatic reactions can benefit from the immobilization, however, the final choice to use them in immobilized form depends on the economic evaluation of costs associated with their use versus benefits obtained in the process. It can be concluded that the benefits are rather significant, since the use of immobilized enzymes in industry is increasing.