Topic 2 and 3
Movement in and out and Enzymes
Human biology june 2023
4HB
12/11/2022
Topic 2
Movement in and out
+ part 1 enzymes
DR. Nihal Gabr
Movement in and out of the cells
Diffusion
1. Diffusion : random movement of molecules from area of higher concentration ( concentrated solution) to
a solution of lower concentration (diluted solution )
2.Factors affecting rate of diffusion 1.Temperature …increase kinetic energy
2.Steepness of the gradient
3.Thickness of the membrane
-
10
Difference = 90
100
4.Size of molecule ….starch / sucrose too large molecules
Difference =20
that can’t diffuse through a membrane , maltose / glucose
200 180 diffuse .
5. Surface area to volume ratio
ammustu 2: 1
3:1
6:1
S.A = Lx W
Total S.A = L. X W x 6
Osmosis
Net movement of water molecules from diluted solution ( ……..) which has higher water potential to concentrated
1
solution (….) which has a lowerv water potential through a partially permeable membrane
2 Then mention the CHANGE observed
Size
Mass -
Volume Water level
Shrinkage Swell /
Remain the Importance of osmosis to human
g
Crenated burst
same size
1. Allow movement of water molecules between
my cells , for metabolic reactions / enzymatic
activity
2. To keep the concentrations of the cell
content constant .
Active transport
Movement of molecules from area of low concentration to area of high concentration against
the concentration gradient using energy from ATP from respiration through carrier proteins
Topic 3 Enzymes
Mechanism of action
Biological catalyst that speed up chemical reaction without itself being changed
Protein in nature
With specific shape of active site Key
Complementary to a substrate
Fit together to form ESC ........
---
_
---
-
~
S
By lock and key mechanism.
O -
-....-
LOCK
Factors affecting enzyme activity
1. Temperature Lev Hydrogen bonds
Enzymatic activity li
2. PH - Maintain shape of the active site
3. Substrate concentration / volume Upon changing pH / temperature
Rate of reaction away from optimum
4. Enzymes concentration / volume
5. Inhibitors a
seemli Hydrogen bonds will start to break
down
Enzyme denature …losing shape
of active site …so active site
Temperature …optimum + near body temperature becomes no longer complementary
to substrate so no ESCs formed
1. Temperature
Explain :
B 1. At 40C : optimum temperature at which the
enzyme works fastest ( near body temperature )
&.
Enzyme
2. Above 40C …..the enzyme will start to
activity Steep decrease
denature
The hydrogen bonds holding the active site
starts to break
Casuing the change in shape of active site
becoming no longer complementary to
substrate , so can’t fit together , so no Enzyme
Temp/ C '
substrate complex
In all enzyme question…..substrate , enzyme ( name) + break down + name product
2.pH
pH =7 is optimum
M
Far away from optimum the enzyme denature where the hydrogen
Enzyme
bonds holding the shape of active site broken down
activity
……………….etc.
7 PA
13/ 11/ 2022
Part 2
Enzymes inhibitors
Immobilised enzymes
Dr. Nihal Gabr
3. Substrate concentration
Limiting factor ….factor present
Rate of B in short supply , so limit the rate of
reaction reaction
~
A Boog
Substrate concentration is
longer a limiting factor
Enzyme concentration is a
Substrate is limiting factor
limiting factor
Substrate concentration
Study
A… The substrate concentration is the limiting factor. As the substrate concentration increases, the
number of collision between enzyme and substrate increase ..so more enzyme substrate
complex ..so more active sites become become occupies so more ESC..so rate of reaction
increase
B……… levels off , where at high substrate concentration , substrate becomes no longer limiting
factor . Where all active sites are occupied so rate of reaction reaches to maximum …so enzyme
concentration becomes limiting factor
4. Inhibitors A. Competitive inhibitor 1. Competitive inhibitor has similar shape to the substrate.
Complementary to shape to the active site
2. So compete with substrate on the active site / so fit and
bind to the active site .
Following the lock and key mechanism ( which includes the
enzymes is the lock and the substrate is the key)
3. So competitive inhibitor stop the key ( substrate ) from
binding to the lock ( enzyme ) .
V max
4. So slowing down the rate of reaction at low substrate
F
Rate of Without inhibitor concentration .
reaction
With the
competitive V max ….point at which all active sites are saturated ( where the
inhibitor enzyme concentration is the limiting factor )
Substrata concentration
Describe the effect of competitive inhibitor on the enzyme activity :
https://youtu.be/rCVRC-AQ54M
- at lower substrate concentration
Lower rate of reaction with inhibitor has larger effect
- so it takes longer time to reach to the maximum rate of reaction
- at higher substrate concentration , the inhibitor has no effect .
+ data .
Read
Oxidation
Methanol ……………………formic acid + formaldehyde
Enzyme
Solution
Ethanol
Competitive inhibitor ( antidote )
Inhibit oxidation of methanol
B. Non Competitive inhibitor
1. Non competitive inhibitor bind to another site other than active site
Causing a change in the shape of active site
So substrate can no longer bind to the active site
of my
So fewer ESCs
So slow down the rate of reaction
At higher substrate concentration …no effect as the substrate can no longer bind to the active site .
- ~
Same enzyme concentration
Inhibitor X
Keep increasing substrate concentration
t
Reached V max Cant reach V max
Competitive Non competitive
How to figure out whether this reaction is inhibited by competitive or non competitive inhibitor ?
Get two beakers , add same concentration of enzymes , keep at same temp and pH
Add in one with inhibitor and the other without .
Then keep increasing the substrate concentration and observe :
1. Lower rate of reaction at low substrate concentration , showing that inhibitor has a larger effect at low
substrate concentration .
Then at higher substrate concentration , the inhibitor will show no effect and ythe enzyme will reach to
maximum rate of reaction ( vmax)
SO THIS IS A COMPETITIVE INHIBITOR
2. Any increase in substrate concentration will not show effect on rate of reaction ( action of inhibitor)
because the substrate cant fit in the active site
So THIS IS A NON COMPETITIVE INHIBITOR .
Competitive Non competitive
Reversible Irreversible
Has similar shape of substrate X
Complementary shape to active site X
Bind to active site Bind to another site other than the active site
Can reach to ( maximum rate ) by increasing Slow down and doesn’t reach to maximum rate by
substrate concentration increasing substrate concentration
Immobilised enzymes Read
go
Advantages of using immobilised enzymes :
1. Recycle enzyme …so can be used more than once
2. No contamination of the product with enzymes
3. Thermostability ( protect the enzyme from changes in temperature) and protect from changes in pH
4. You can prepare a large column , so will expose the substrate to larger enzyme concentration so increasing
rate of reaction .
5- investigation
Notice : Beads are
How to prepare lactose-reduced milk: formed by sodium
2- Add the alginate 3- then the mixture is poured alginate , having
3
enzyme mixture to through a sieve to separate the lactase trapped in
2 100cm3of 1.5% calcium beads from the solution. them.
chloride solution drop by Beads are washed with distilled
drop water.
1- Mix 2cm3 of enzyme
with 8cm3 2% sodium
r
- insoluble calcium
alginate solution then alignate beads are
ab
mix throughly using a formed. the beads are
stirring rod. left to set for a 5
4
minutes. 4- pack
the beads
in a clean
6- The milk leaving the syringe is
tested using glucose test strips. After a
while, glucose will be present in the
milk that has passed through the
column of beads. As a Control, the
milk is tested before treatment – it
5- pour
lG 10cm3
syringe.
iha
some milk
will not contain any glucose..
gently over Nylon
the beads in gauze.
9 5
the syringe.
.N
Safety precautions:
Wear eye protection and avoid skin contacts
with the lactase, alignate solution and gel.
Don’t taste the whole milk or the lactose free
Dr
milk.
1. Suggest why you need to wash the beads before putting them into the syringe barrel?
To remove any calcium chloride from them, which might affect the reaction.
2. What was the purpose of the piece pf nylon gauze in the syringe barrel?
To hold the beads in place, so they don't fall into the syringe and block it.
3. How quickly did the liquid move through the immobilised enzyme?could you speed of up?
Might this affect your yield of glucose?
You would expect to get higher yield of glucose if the milk flowed through more slowly, as it would remain
in contact with the enzyme for longer.
4. At stage 2, it is important that the tip of the syringe does not touch the calcium chloride
solution, or it will block with gel/ beads)
040
Examples 01093850599
1. Lactose free milk
Some people they lack lactase enzyme in their small intestine.
So lactose milk ( disaccharide) is not broken to glucose and galactose upon
consumption of any milk product ,
So cant be absorbed from small intestine into blood
Suffere from lactose intolerance
Abdominal bloating , flatulence , nausea
Taking pills containing the Lactose free products using
lactase ezyme immobilised enzymes
Damaged by acidity of the stomach
+ expensive enzyme ( lactase ) .
2. Breaking down sucrose to produce invert syrup
Sucrose ………invertase enzyme ………………….glucose + fructose
Produced by yeast cells
Trapped in alginate beads
Monosaccharides
Sweeter than sucrose
+ low calorie
3. Testing stripes for testing glucose in urine:
Advantages
This method
is specific Immobilised enzyme 1. Glucose oxidase
for glucose 2. Peroxidase
( not
affected by
other
sugars )
Urine pass over the cardboard containing the immobilised enzymes
Glucose bind ti immobilised enzyme ( glucose oxidase ) to be oxidised into gluconic acid and H2O2
Glucose oxidase
GLUCOSE + O2………………………………GLUCONIC ACID + H2O2
H2O2 ….oxidise a colorless organic substance ( XH2) to colored one using PEROXIDASE enzyme
H2O2 + XH2………………….2H2O + X colored compound
The higher the glucose concentration, the darker the color
