Detergent enzymes purification processes

In biochemical assays, additives such as detergents, DMSO, urea, BSA, and glycerol are commonly used to improve reaction performances and enzyme stability. However, these additives also act as crystallization disturbing agents preventing the formation of optimal crystals for the MALDI process. Analytical sensitivity and mass accuracy can be affected. The challenge is to develop bioassays that can perform optimally without crystallization disturbing additives. Often, it is necessary to use elaborate purification processes prior to analysis. 

The product quality demands are highly dependent on the application of the enzyme. Detergent enzymes typically have requirements for their physical appearance (color, odor) and also for compatibility with the detergent. Impurities related to the process (e.g., the presence of unfermented sugar from the media) and microorganism (e.g., metabolites produced during the fermentation) can influence the quality of the product. Dealing with these issues typically requires addition of one or more purification steps in the process. 

Detergent enzymes are produced by bacterial strains, conditioned to resist alkaline pH s and therefore to produce alkaline-resistant enzymes. Moreover, they are exocellular enzymes, secreted by the bacteria into the surrounding medium. Thus, they can be isolated without breaking the bacterial cells. That makes the purification process easier and less costly. These bacterial proteases are less specific and will degrade almost all kinds of proteins. 

After recovery and purification of the enzymes, the third important process step is to make a final formulation of the enzymes. The enzyme concentrate, whether it is present in an aqueous solution or in a dried form, has to be transformed into a stable form that fulfils the detergent producers requirements. Obviously, the chosen formulation needs to be compatible with the detergent and a range of different quality parameters should be defined. 

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