Lab 3 (enzyme action) Bio 1

enzymes importance

accelerate and steer chemical processes in organisms

The active site of an enzyme is (3)

catalytically active region of an enzyme, shaped to fit a specific substrate, destroyed if the shape changes drastically

how do high temps and strong acids affect an enzymes shape

cause gross untwisting and disorganization of the structure which destroys the active site and abolishing the enzymes catalytic power

importance of enzymes configuration

chemically reactive groupings make up the catalytic active region (active site)

ONPG (colorless) + lactase =

galactose and ONP (yellow)

lactose + lactase =

galactose and glucose

2 examples of agents that can alter an enzymes shape

high temperatures, strong acids

How does a spectophotometer help?

if less light goes through the solution there must be lots of ONPG since it means a yellow color which causes more absorbance of light

what is the roles of Na2CO3 in activity 1 and 2

inactivate enzyme

what stops the production of galactose and ONP?

sodium carbonate (Na2CO3)

how can we determine how much enzyme is present in a sample of unknown concentration?

using the relationship between enzyme concentration and ONP production (amount vs activity --> standard curve)

Lactase catalyzes the conversion of

ONPG into galactose and ONP

3 steps of making a blank tube in activity 2 (produce a standard curve of enzyme amount vs activity)

(1) 1 ml of .6 units/ml enzyme solution (2) 3 ml of Na2CO3 and mix (3) 1 ml ONPG and mix

3 steps of making a blank tube in activities 1 and 3(investigate the activity of the enzyme lactase / temperature on enzyme activity )

(1) 1 ml of lactase (2) 3 ml of Na2CO3 and mix (3) 1 ml ONPG and mix

what is lactose

A disaccharide made up of glucose and galactose

Why do we add NaCO3 to our tubes?

To stop enzymatic production of ONP

what is lactase

enzyme that catalyzes the digestion of lactose

Why is ONP useful in this experiment?

it's yellow so we can measure how much ONP is produced by the intensity of yellow in the mixture

what happens when ONPG binds to the lactase active site?

its broken down into galactose and ONP

In this lab, higher enzymatic activity will cause:

more ONP production and higher absorbance

Why is ONPG an ideal substrate for lactase

they are similar in structure so it can readily bind to the lactase active site

what are 3 things modified in the experiments?

time, enzyme concentration, temperature

why is lactase important for lactose

to be used as the glycosidic bonds must be