Salivary Amylase

Activity of Salivary Amylase
Domingo, Guray, Hugo, Lorenzo, Mohammad Isa

Intro
Because everything has a start

Catalysis

 The process of increasing the rate of reaction with the use of a catalyst.
 Catalyst – any substance that increases rate of reaction upon addition to a certain reaction

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Enzymes

 Act on substrates in a reaction
 Highly specific
 Breaks down complex macromolecules, synthesizes compounds essential for the cell
 Active site
 Enzyme-substrate complex
 Speeds up reaction rates

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http://www.cas.muohio.edu/~wilsonkg/old/gene2005/syllabus_F03_23.jpg
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Enzymes

 Require cofactors for activity
 Classified according to the types of reaction they catalyze

– Oxidoreductase
– Transferase
– Hydrolase
– Lyase
– Isomerase
– Ligase

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Amylase

 An enzyme that breaks down starch into oligosaccharides through hydrolysis  Secreted by the human’s parotid glands and the pancreas
 α-Amylase
 β-Amylase
 γ-Amylase

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Factors that may affect catalysis rates

 Temperature
 pH
 Enzyme concentration
 Amount of substrate

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Materials and Methods

Solution Preparation

Saliva was collected.
1 ml of saliva was diluted to 10 ml with distilled water.
 10 % salivary amylase solution

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Estimation of salivary amylase activity

A mixture of 5.0 mL 1% starch, 2 mL 1% NaCl solution and 2 mL phosphate buffer put in a test tube and then placed in a water bath
At 38oC, 1 mL salivary enzyme solution added to the solution.
A drop from the digestion mixture mixed with 1 drop of iodine for every minute.  Achromic point was determined.

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Effect of enzyme concentration

The salivary amylase solution diluted to five lower concentrations: 2.5%,
2.0%, 1.5%, 1.0%, 0.75% and 0.5%.
 The same procedure done as previous using 1% concentration of starch solution. Reaction rates observed