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Lab #5 - PHS 2301 BIOMED I LAB 5 Enzyme Kinetics Previous...

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LAB 5 Enzyme Kinetics PHS 2301 - BIOMED I
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We studied Dissociation of p-Nitrophenol (p-NP) Least in an acidic pH – colorless solution Highest in alkaline pH – yellow solution ( λ max at 400nm ) Enzyme activity Optimal conditions (pH & Temp) Enzyme substrates Previous Labs
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Objectives To study the enzyme kinetics of Acid Phosphatase using the substrate p-nitrophenol phosphate disodium salt (p-NPPO4Na 2 ) To determine unknown concentrations using a standard curve To determine the Michaelis constant, K m and Maximum velocity, V max of Acid Phosphatase enzyme
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ENZYMES - APPLICATIONS Diseases – Important diagnostic tools Prostatic acid phosphatase - prostate cancer BioTechnology – Manipulate genes Restriction digestion enzymes – Cut DNA into fragments (We will learn more in the next labs) Drug metabolism – P450 enzymes Used in food processing and in chemical industry
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ENZYMES Enzymes are proteins that catalyze in biological reactions Substrate (S) + Enzyme (E) ES complex E + Products (P)
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ENZYME KINETICS When substrate concentration is low, Not all of enzyme is bound to substrate (Free enzyme is still available) As the substrate concentration is increased, more of the
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