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ERT 314
BIOREACTOR SYSTEM
COMPARISON
ON
ENZYMATIC
ACTIVITY BETWEEN IMMOBILIZED AND
FREE CELL REACTOR FOR ETHANOL
PRODUCTION IN BATCH CULTURE
DATE OF EXPERIMENT
22ND APRIL 2015
PREPARED FOR
DR. KU SYAHIDAH BINTI KU ISMAIL
GROUP MEMBERS
(B4)
Experiment 5
Comparison Enzymatic Activity between Immobilized and Free Cell Reactor for
Ethanol Production in Batch Culture
1.0 Objectives
1.1 To comparison enzymatic activity between immobilized and free cell reactor for
ethanol production in batch culture
1.2 To understand the principles of cell immobilization by using gel entrapment
method in alginate gel
1.3 To analyze the effect of immobilization on ethanol production
2.0 Introduction
Malt Extract
3.2
Peptone
3.3
Dextrose (D-glucose)
3.4
3.4
3.6
3.7
3.8
Distilled Water
3.9
Tap water
3.10
Ice
3.11
DNS
3.12
3.13
Pipette
3.14
Beaker
3.15
Autoclave
3.16
Incubator
3.17
Chemical balance
3.18
Spectrophotometer
3.19
3.20
3.21
Retort Stand
3.22
Syringes
4.0 Procedures
4.1
Inoculation of yeast
1.
0.8% (w/v) malt extract, 1.0% (w/v) peptone and 2 % (w/v) dextrose
(D-glucose) is prepared for a working volume of 100ml.
This solution is put in a 100 ml Erlenmeyer flask.
The flask is cotton-plugged and covered with aluminium foil.
The medium is sterilized in batch autoclave at 1210C for 15 minutes.
2.
3.
4.
5.
before inoculation.
6. 100ml of the medium is measured and poured it into a conical flask.
7. 2 loops of yeast is scraped from agar plate and is inoculated it into the
medium and are left in the incubator to be shaken overnight.
5% (v/v) of yeast culture is pipette into the flask respectively.
The fermentation broth is incubated in incubator shaker at 150rpm.
8.
9.
4.2
4.3
Time
0
Immobilised
Enzyme
15 min
30 min
45 min
60 min
Glucose
(g/L)
Glucose
(g/L)
Free Enzyme
Expected Graph
Comparison in term of Absorbance between Immobilized Yeast Cell and Free Yeast Cell
over Time
12
10
8
Absorbance, nm
6
4
Time, min
10
12