Professional Documents
Culture Documents
Biofuel Enzyme Kit Protocol
Biofuel Enzyme Kit Protocol
Highlights:
Using a
colorimetric
substrate to
track reaction
rate
cellobiose
p-nitrophenyl glucopyranoside
Cellobiase
breakdown of pnitrophenyl
glucopyranoside
glucose
p-nitrophenol
Basic
conditions
Clear
Yellow
Biofuel Enzyme
Kit
Procedure
Overview
Example of
Standards'
Absorbance
Readings
Standard
S1
Amount of
p-nitrophenol
(nmol)
0
Absorbance
410 nm
0
S2
12.5
0.2
S3
25
0.4
S4
50
0.8
S5
100
1.6
Qualitative
Determination
of Amount of
Product
Formed
Quantitative
Determination of
p-nitrophenol
Amount
Read Samples
Analyze Results
Quantitative
Determination
of p-nitrophenol
Amount
Calculating
initial reaction
rate with and
without an
enzyme
present
Amount of p-nitrophenol
produced (nmol)
Time (min)
50 nmol - 0 nmol
4 min - 0 min
= 12.5 nmol/min
Conditions
affecting
reaction rate
pH
Temperature
Substrate Concentration
Enzyme Concentration
Effects of pH
Prepare and
run reactions
Calculating
initial
reaction rate
at different
pH values
Amount of p-nitrophenol
produced (nmol)
Time (min)
Further
activities
included in
the kit