Microbiological Testing of Enzymes

What are Enzymes?

Enzymes are specialized proteins that accelerate the rates of chemical reactions by lowering the activation energy; the substrates are converted into products.  There are numerous different types of commercially available enzymes such as protease, amylase, cellulose and lipase.

Where are they used?

Many different industries use enzymes in the production of their products.  They range from animal nutrition, brewing industry, food processing, detergents, dairy industry, paper industry, biofuels, contact lens cleaners, photographic industry, carbohydrate processing and biochemicals.  Enzymes are also an integral part of the pharmaceutical and Nutraceutical industry.  In laundry detergents enzymes that degrade proteins causing stains, such as those found in grass stains, red wine and soil can be used.

Lipases are another useful class of enzymes that can be used to dissolve fat stains and clean grease traps or other fat-based cleaning applications.  Nutritional enzymes have risen to prominence during the past several years, driven by customers that would like to improve their digestive health. The biggest category for supplemental enzymes is still human digestion, an area that has become increasingly more important for maintaining optimum health. Supplementation of single enzymes, such as lactose to treat lactose intolerance, is probably the most familiar area of enzyme use. However, general dietary supplementation with a broad range of digestive enzymes is becoming increasingly more common. Supplemental enzymes can also be used to enhance the effect of herbs and botanicals. The active component of herbs and other botanicals can remain trapped within the cell wall matrix or may be in a form that is not easily broken down and absorbed. Select enzymes are added to these products to break open certain bonds to free the bioactive compounds.  The application of enzymes technology to pharmaceutical products is also a growing field.

How do they Work?

Amylases in the detergents can help by aiding in the breakdown of starch stains.  Also in the biofuel industry cellulases are used to break down cellulose into sugars that can be fermented, and then further processed to ethanol.  Trypsin is used in baby food to predigest the food for the babies.   Proteases can also be used in contact lens cleaners to remove proteins on the lens.  The figure below show a schematic diagram of how enzymes work:

Most enzymes require microbiological testing similar to other industrial products.   BioLumix has an easy method to complete this goal.  The versatility of the BioLumix system allows for a wide range of enzymes to be tested for a broad array of microorganisms; such as Total Aerobic Count, E. coli, Salmonella, Staphylococcus aureus, Pseudomonas aeruginosa, Lactic Acid Bacteria, Coliforms, Enterobacteriaceae, and Yeast and Molds. Liquid products allow a direct sample to be pipetted directly into the vial, while solid samples require a dilution step first.

The BioLumix Rapid Automated Method:

Enzymes traditionally can be very difficult to test due to their unique properties.  The two most common difficulties when working with enzymes is solubility and pH levels of the product.  When using the BioLumix system, pH is adjusted once the product has first been diluted at 1:10 in TSB or buffer during the preparation before the sample is placed into the vial.  Solubility issues can be solved in a few different ways. One way is by allowing the product to sit in solution and give it time to breakdown, and another way it to warm the product in a water bath or an incubator.

During enzyme testing at BioLumix, on plates the enzyme created a precipitate that looked very similar to colonies, which could lead to difficulties in reading the plate.  The plate needed to be  sub-cultured to determine if it was growth or just the precipitate.  The precipitate did not interfere with the BioLumix vials and provided a clear answer.  Additionally, the results were obtained two days sooner.  An oil based enzyme can also cause difficulties when using traditional plating methods versus the BioLumix system.  In the BioLumix system the oil does not have an effect on the result and can give a definitive answer.  While on plates, the oils can sometimes appear on top of the agar and give the appearance of colonies.

Another less common issue is gas production by the product when it is mixed with a liquid.  If the product is known to release gas the easy solution is to allow the product to off gas before adding it to the appropriate vial.  Then after an hour of being in the liquid, the sample can be stomached or mixed again to release the remaining gas.  While in traditional methods, the gas production can give false positive results in tests with Durham tubes, which would then require additional days to complete.

BioLumix Advantage:

The BioLumix system is designed to accelerate product release with a simplified, automated approach. This yields fast, accurate, real-time results while reducing costs. The BioLumix offers real-time results of contaminated samples saving hours, possibly days. Completed Total Aerobic Count, Coliform or Enterobacteriaceae results are obtained within 18-22 hours; Yeast and Mold results are obtained within 48 hours; the objectionable organisms can be detected within 24 hours after pre-enrichment. Also these assays are simpler to perform than the standard methods saving time, labor and money. The system is unaffected by product interference, delivering accurate results with faster product release.  When referring to batches of product that will be released days early due to the BioLumix system, which allows the storage and production less time to run and hastens the turnaround time of the product, the overall process of enzyme production will decrease.

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