The Effects of varying p-Nitrophenyl Phosphate concentrations on Alkaline Phosphatase Activity Nathan Overholt Louis Settembrino March 5, 2002 Abstract: The enzyme Alkaline Phosphatase was set up in solutions of varying p-nitrophenyl phosphate concentrations. A fixed time assay was used to find the Michaelis constant and Vmax. All solutions were incubated at 30 C° for 30 mins. Absorbances were measured at 405 nm, and the Km was found to be 0.002 M (pNPP) and the Vmax to be 0.433 A/min.
Introduction: Alkaline Phosphatase: Alkaline Phosphatase is an enzyme that comes from liver, bone, placenta, and intestine. This enzyme hydrolyses phosphate esters, is secreted into the serum by osteoblasts and is used as a diagnostic marker for increased metabolic activity. Growing bones need alkaline phosphatase. Any condition of bone growth will cause alkaline phosphatase levels to rise. The condition may be normal, such as a childhood growth spurt or the healing of a broken bone; or the condition may be a disease, such as bone cancer, Paget's disease, or rickets. During pregnancy, alkaline phosphatase is made by the placenta and leaks into the mother's bloodstream. This is normal. Some tumors, however, start production of the same kind of alkaline phosphatase produced by the placenta. These tumors are called germ cell tumors and include testicular cancer and certain brain tumors. (Nordensen, 2002) Healthy livers usually carry alkaline phosphatase away along with other harmful substances out through the bile duct. If the liver becomes diseased in some sort, then the bile duct may become blocked producing large amounts of built up alkaline phosphatase and eventually this leaks out into the bloodstream. There are different forms of alkaline phosphatase called isoenzymes. The determination of the particular isoenzyme will determine the target problem area. (Sorensen, 2002) Enzymes: Enzymes are catalysts, meaning, they help a reaction proceed without being consumed. The substrate...
Cited: Delaunay J, Fischer S, Piau JP, Tortolero M, Schapira G. Apr. 2, 1993, Properties of a membrane-bound phosphatase activity in normal and abnormal red blood cells. (Abstract) Nordenson, Nancy J. 2002. Alkaline Phosphatase Test.
http://www.ahealthyme.com/topic/topic100586408 Ozegowski JH, Muller PJ. Dec. 25, 1984, Metabolism of phosphate-limited Streptomyces cultures. I. Purification and characterization of alkaline phosphatase produced by Streptomyces hygroscopicus. (Abstract) Sorensen, Ralph A. Spring 2002, Alkaline Phosphatase Activity, http://www.gettysburg.edu/~rsorense/Celllab02/ap.htm
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