IA on efficiency of immobilized enzyme amylase (fungal extract) in alginate beads, in starch hydrolysis.

1. Efficiency of immobilized enzyme amylase (fungal extract) in alginate beads, in starch hydrolysis
Sodium alginate reacts with calcium chloride to form calcium alginate
These beads can be used to immobilise enzymes so it can be recycle/reuse
Encapsulation of drugs/enzymes can be done using alginate beads.
3% (w/v) sodium alginate and 2% (w/v) calcium chloride investigated.
Sodium alginate + calcium chloride
forming alginate beads.
Procedure:
Weigh 3g of sodium alginate (SA) in 100ml water - 3% SA.
Weigh 2g calcium chloride (CC) in 100ml water – 2% CC.
0.05g amylase powder mixed with 3ml of 3% SA.
Pipette 1ml 3% SA/amylase and drop it into 2%CC, making 10 beads in total.
10 alginate beads added to starch/I2 mix in next slide.
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3% (w/v) SA used
3% (w/v) sodium alginate used
10 beads of same size taken out Beads measured with colorimeter
3% SA with 0.05g enzyme + 2% CC
Pipette tip used to form the beads

2. Procedure:
1% (w/v) starch – 1g in 100ml water
1% (w/v) iodine – 1 g in 100ml water
1% (v/v) Enzyme amylase obtained from fungal extract
1ml starch + 20ul I2 was added into a cuvette. (Blue black)
10 alginate beads with enzymes immobilized was added into cuvette.
Ini and final after abs taken over 3 mins.
Rate of hydrolysis – decrease in abs over time. (Day 1)
Keep the beads in fridge and measure the abs for Day 2.
Starch react with I2 to form a blue black product.
Blue black colorization can be monitored using visible spec.
Enzyme amylase will breakdown starch to maltose.
As starch is broken down, I2 wont be able to bind with starch.
Resulting in colorless product.
Rate of starch hydrolysis – measured by change in blue black over time (3mins).
Starch + I2 + enzyme → maltose + simple sugars
Reaction mechanism
Starch + I2 (Blue black) I2 (yellow)
10 beads of same size taken out Beads added to cuvette, and abs taken
Efficiency of immobilized enzyme amylase (fungal extract) in alginate beads, in starch hydrolysis

3. Go to expt – press calibrate Insert cuvette (blue black) and press collect
Insert a blank containing water
Press stop, and click on rainbow icon. Select abs vs time. λ max at 614nm will
be automatically chosen
Abs vs time
Efficiency of immobilized enzyme amylase (fungal extract) in alginate beads, in starch hydrolysis

4. 3% SA added to 2% CC seems
to be the best
1% SA + 1% CC 2% SA + 2% CC 3% SA + 2% CC
Procedure:
Weigh 3g of sodium alginate (SA) in 100ml water - 3% SA.
Weigh 2g calcium chloride (CC) in 100ml water – 2% CC.
0.05g amylase powder mixed with 3ml of 3% SA.
Pipette 1ml 3% SA/amylase and drop into 2%CC, making 10 beads in total.
Ini and final abs taken over 3 mins period.
Rate of hydrolysis – decrease in abs over 3 mins (Day 1).
Keep the beads in fridge and measure abs again for Day 2.
Beads able to breakdown starch after 1 day, with 79% efficiency
Efficiency of immobilized enzyme amylase (fungal extract) in alginate beads, in starch hydrolysis
Day Ini abs Final abs Rate/
absmin-1
Efficiency
%
1 2.067 0.921 0.382 100
2 2.381 1.470 0.303 79
0
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1 2
Efficiency
Day
Day vs Efficiency