Enzymes are therefore dependent on substrate concentration, sil microorganisms’ metabolic ability, sil environmental conditions such as temperature and pH and the number of microorganisms present in the soil (Lichtfouse, 228). Akaline Phosphatase is an example of extracellular enzyme produced by many soil microorganisms and exported out into the soil solution. Te main function is to eliminate the phosphate molecule from the organic compounds such as nucleic acids and phospholipids. Tis makes phosphate soluble hence can be easily absorbed by the cells. Tis is very important because phosphate is often limiting nutrient for microbial growth in the soil.
Masurements of soil enzyme activities are therefore useful indicators of soil biological activity. Mny studies have also referred to soil enzyme activity as an index for soil health and quality of land (Verchot and Borelli, 629). Ezyme activity has also been used in studies that investigate the impact of human activities on the biochemical cycles in ecosystem. Te purpose of this study is to measure the amount of active enzyme in the soil samples by applying chromogenic substrate assay. Te principle Alkaline Phosphatase assay is that alkaline phosphatise catalyzes the conversion of a colourless para-nitrophenol phosphate to para-nitrophenol which has a bright yellow colour.
Te rate of enzyme activity in this experiment is calculated from the spectrophotometer readings from the amount of para-nitrophenol formed. Te basic enzyme assay applied in this study is the addition of known amount of soil to a solution containing a standard concentration of substrate then measuring the rate at which substrate is converted to a product while keeping enzyme environmental factors such as temperature, inic and pH constant.
Te results of this study will also be standardized by calculating the dry weight of the soils. Te materials used in this experiment were categorized s equipment, mdia and reagents and supplies. Euipment included: 370C incubator, Blance, Auminium weighing dishes, sissors, 16x100mm test tubes, 5ml pipettes and pumps, 440nm Spectrophotometer, dying oven and benchtop centrifuge. Mdia and reagents included: Bffer of pH 10, 05M CaCl2, pra-nitrophenol Phosphate (PNPP) in buffer which served as the test solution and 2mM p-nitrophenol. Spplies included: Pastic flowerpots, mrkers, dy, seved flter paper and yeast extract.
Al the supplies, mdia and...
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