Analysis of natural microbial enzyme activities found in a proprietary fermentation product
Abstract
Chemical processes are harmful to people, animals, and the environment and are being replaced with more environmentally friendly strategies. Biocatalysts, such as enzymes, are safer for everyday use. Enzymes have multiple benefits that make them extremely attractive. Enzymes are currently used in the antibiotic industry, food industry and textile industry. Additionally, enzymes work effectively and efficiently at breaking down various inorganic and organic materials. This includes paper, human waste and particles regularly found in stains. Enzymes are now commonly found as components in detergents and industrial cleaners as they reduce time, water and energy consumption. The most common type of enzymes employed in the cleaning industry are hydrolases. The three most common hydrolase enzymes found in cleaning products are lipases, proteases and invertases. Lipases break down triacylglycerols into glycerol and free fatty acids. Lipases are important in the cleaning industry as they break down animal and vegetable fat and oil stains as well as aid in the removal of soil stains. Proteases break down proteins and peptides at their amide bonds. Proteases are imperative in the cleaning industry due to their ability to break down many types of stains. Invertases are the third hydrolase enzyme used in the cleaning process. Invertases break down sucrose into fructose and glucose. Invertases are used in cleaning products since sugars are common in stains. The studies performed in this thesis were designed to determine if/which enzymes were components of a proprietary microbial fermentation product manufactured by a local company. Proteins were detected in the product suggesting the potential for enzymatic activity. Assays were designed to test for lipase, protease, and invertase activity in "fermentation broths" obtained directly from the manufacturer and also "starter cultures" produced from inoculation of fresh proprietary material (organism). Lipase activity was detected in starter culture material via both colorimetric and agar plate methods. Protease activity was found in a colorimetric assay for some fermentation broths but was at the lower limit of detection. Protease activity was also found in starter cultures using an agar plate method. Invertase activity was detected via a colorimetric assay that measures the level of free glucose. Invertase activity was found in both fermentation broth and starter culture materials. Invertase activity was quantifiable and could potentially be used as a measure of quality control during production of the product. In summary, enzymatic activities were detected in one or both of the materials suggesting that one or more of these enzymes are an active component in the proprietary cleaning product.
Degree
M.S.
Advisors
Visalli, Purdue University.
Subject Area
Biology|Biochemistry
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