Biofuel Enzyme Kit

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Overview

Kit contains sufficient materials for 8 student workstations (2–4 students per workstation).
Enzyme, cellobiase, 400 µl	1 vial
Substrate, p-nitrophenyl glucopyranoside, 90 mg	1 vial
Standard, p-nitrophenol (1 mM, 4 ml)	1 bottle
2x stop solution, 100 ml	1 bottle
10x resuspension buffer, 50 ml	1 bottle
Extraction buffer, 50 ml	1 bottle
Disposable plastic transfer pipets	40
1.5 ml microcentrifuge tubes	90
15 ml conical tubes	50
Disposable cuvettes	100
Instruction manual	1
Required Accessories Not Included in Kit
Instructor setup or lab equipment	Quantity Needed per Kit
500 ml bottle for 1x resuspension buffer	1
100 ml bottle for high concentration enzyme	1
100 ml bottle for low concentration enzyme	1
100 ml bottle for 3 mM substrate	1
150 ml bottle for 1.5 mM substrate	1
50 ml tubes or bottles for standards	5
Serological pipettor and pipet or Graduated cylinders to measure volumes 4–450 ml	1
Deionized or distilled water	1,000 ml
Lab tape (not Scotch tape)	1
Student workstation (4 students) All Activities	Quantity Needed per Station
Marking pens	8
Timers or stopwatches	8
Activity 2: Effects of Temperature
Beakers with 37°C water	8
Beakers or ice buckets with chipped ice	8
Thermometers	8
Activity 6: Mushroom Extracts
Beakers	16
Mortar and pestles	8
Balance	1
Weight boats or weigh paper	1
Optional Accessories	Quantity
100–1,000 µl adjustable micropipets	8
100–1,000 µl pipet tips	8
SmartSpec™ Plus spectrophotometer or similar	1
Water bath, 120 V	1
Digital dry bath, 120 V	1
DyNA Chill cooler	1
Mini incubation oven, 120 V	1
Mini centrifuge, 120 V	1

Biofuel Enzyme Kit Flowchart

Key Kit Features

Aligns with AP Biology Lab 2
Guides instruction on enzyme kinetics and biofuel energy sources
Contains no caustic reagents
Enables both qualitative and quantitative measurements of reactions
Complete core activity in one 45-minute session
5 additional enrichment activities

Curriculum Fit

Carbon Cycle

A carbon cycle with the inclusion of biofuel production and use. Plants remove carbon dioxide from the atmosphere and in the presence of sunlight can convert it to complex polysaccharides including cellulose. Plants such as switchgrasses and poplar trees can be harvested and processed in a cellulosic ethanol plant to produce ethanol to fuel planes, cars, and trucks. However, the tractors and other vehicles used to process the switchgrasses, trees, and plants and the vehicles running on cellulosic ethanol still produce carbon dioxide. Carbon dioxide also comes from respiration of animals, burning of fossil fuels, and breakdown of organic matter by microbes. Ideally, carbon dioxide produced by the manufacture and processing of biofuels will be reabsorbed by plants used to make the cellulosic ethanol. This is not the case with fossil fuel usage. The fossil fuels only contribute to carbon dioxide production.

Reveal the power of enzyme kinetics by illustrating their function through the real-world application of biofuel production. The Biofuel Enzyme Kit measures the enzymatic activity of cellobiase (part of the cellulase family) and identifies the optimal conditions for the enzyme. The reaction of cellobiase breaking down cellobiose is important in the process of making cellulosic ethanol, which is an efficient, more sustainable fuel to replace petroleum. Students test and calculate the conversion rate of a sugar substrate (p–nitrophenyl glucopyranoside) to p–nitrophenol and glucose in the presence or absence of cellobiase.

Features and Benefits

Aligns with AP Biology Lab 2
Guides instruction on enzyme kinetics and biofuel energy sources
Contains no caustic reagents
Enables both qualitative and quantitative measurements of reactions
Provides additional enrichment activities

After establishing the reaction rate in the presence of the enzyme, various conditions can be tested. Students can perform the following independent inquiries:

Determine the effect of temperature on the reaction rate
Determine the effect of pH on the reaction rate
Determine the effect of enzyme concentration on the reaction rate
Determine the effect of substrate concentration on the reaction rate
Test the ability of mushroom extracts to increase the reaction rate

Background Information

Cellulosic ethanol is a biofuel derived from plant matter. Cellulose is a polysaccharide found in the cell walls of plants. The breakdown of cellulose into sugar is a multistep process that is facilitated by a family of enzymes called cellulases. Each cellulase has its own unique role in processing cellulose from a long strand of glucoses down to single units that can then undergo microbial fermentation to produce ethanol.

More Information

One 45 min laboratory activity
Provides sufficient materials for eight student workstations, up to four students per workstation

Kit contains sufficient materials for 8 student workstations (2–4 students per workstation)
Enzyme, cellobiase, 400 µl	1
Substrate, p-nitrophenyl glucopyranoside, 90 mg	1
Standard, p-nitrophenol (1 mM, 4 ml)	1
2x stop solution, 100 ml	1
10x resuspension buffer, 50 ml	1
Extraction buffer, 50 ml	1
Disposable plastic transfer pipets	40
1.5 ml microcentrifuge tubes	90
15 ml conical tubes	50
Disposable cuvettes	100
Instruction manual	1

Biofuel Enzyme Kit

166-5035EDU
Enzyme kinetics kit, includes enzyme, substrate, standard, solution, buffers, disposable pipets, tubes, and cuvettes, for 32 students; education use only

List Price:		$158.75
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Number	Description	Options
10017703	Instruction Manual, Biofuel Enzyme Kit, Biotechnology Explorer, Rev B	Please login to download this document
5947	Instruction Manual, Biofuel Enzyme Kit Quick Guide, Biotechnology Explorer, Rev A	Click to download

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