Investigations with phosphatase Paul Beaumont Science & Plants for Schools Homerton College, Cambridge.

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Investigations with phosphatase Paul Beaumont Science & Plants for Schools Homerton College, Cambridge

OCR specification Enzymes – Learning Outcomes: (c) describe and explain the effects of pH, temperature, enzyme concentration and substrate concentration on enzyme action. (d) follow the time course of an enzyme-catalysed reaction by measuring rates of formation of products or rate of disappearance of substrate (e) explain the effects of reversible inhibitors (both competitive and non-competitive) on the rate of enzyme activity. 5.1 Module 2801: Biology Foundation

Phosphatase Phosphatase enzymes from plants: a versatile resource for post-16 courses Meatyard, B. (1999), J. Biol. Educ., 33(2),

Phosphatases Found in wide range of plant and animal tissues Release phosphate groups Phosphate available for ATP and nucleotide synthesis, membrane construction. Two main groups – acid and alkaline

The Assay

Experimental Outline Preparing a standard curve (Part A) Preparing the enzyme extract (Part B) Running the assay (Part C)

PPP = phenolphthalein phosphate (substrate) PP = phenolphthalein (product)

Kit contents

Additional materials

Standard curve Solution Number Volume of PP stock, (cm 3 ) Volume water, (cm 3 ) Volume Na 2 CO 3 (cm 3 ) PP concentration (mol dm -3 ) x x x x x 10 -5

Colorimeter R = ReferenceT = Test Direction of Beam

Standard curve

Germination

Creating the extract

Maceration

Loading centrifuge tube

Centrifugation

Retain supernatant

Running the Assay

Running the assay

Results

Results

Phosphatase Investigations Phosphatase Investigations Effect of temperature Effect of pH Phosphatase levels from different species Phosphatase levels in different locations Effect of phosphate concentration