Biofuel Enzyme Reactions Kit for AP Biology

Biofuel Enzyme Reactions Kit for AP Biology


Explore enzyme kinetics associated with the production of cellulosic ethanol, a biofuel. This ThINQ! Kit provides support for an open or guided inquiry format, enabling a student-driven learning approach. Covers Big idea 2 and 4.

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Enzyme Lab Activity for AP Biology and Biotechnology Classrooms

The Biofuel Enzyme Reactions Kit for AP Biology is a student-driven, inquiry-based kit that characterizes the activity of the cellobiase enzyme. In this ThINQ! Investigation, students will play the role of bioengineer to explore the power of enzyme kinetics and optimize conditions that will maximize the rate of sugar conversion for the production of biofuel.

Features and Benefits

  • Aligns with AP Biology Big Ideas 2 and 4
  • Introduction to enzyme kinetics with real world application to biofuel energy
  • Enables both qualitative and quantitative measurements of reactions
  • Contains no caustic reagents

Classroom Experience

Students first extract cellobiase from mushrooms to discover one way that nature uses enzymes. They then generate questions and hypotheses about enzyme function under different environmental conditions — temperature, pH, and concentrations — that influence enzyme function. Students can take a structured, guided, or open approach to inquiry as they test their hypotheses. Finally, with data in hand, students use evidence to create a list of conditions for optimal enzymatic activity for biofuel production.

Timeline and Contents Overview

The flexible curriculum will fit within a broad range of available laboratory times. A possible arrangement is:

  • Day 1: Pre-lab activity and background review
  • Day 2: Investigation 1
  • Day 3: Investigations 2–5 (divided into groups)
  • Day 4: Investigation 6 and post-lab assessment
  • Day 5: Science Case Study

Background Information

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

A key step in the production of cellulosic ethanol is the breakdown of cellobiose by the cellobiase enzyme. 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.

Select the kit which is right for you and your students:


ThINQ! Biofuel Kit

Classic Biofuel Kit

Standards alignment

AP Biology

General Biology, Biotechnology, CTE

Pedagogical support

Open, guided, and structured inquiry

Structured inquiry

Main learning objectives

Focus on student-driven questions and problem solving

Mastery of technical skills

Initial experiment

Compare reaction rates of mushroom extracts

Focuses on inquiry process

Compare reaction rates with and without purified enzyme

Uses well characterized samples to obtain reproducible results

Kit contents

Materials for 8 student workstations to perform 1 initial experiment and up to 5 additional enrichment activities

Explore features of the Classic Biofuel Enzyme Kit.



Kit contains sufficient materials for 8 student workstations
(2–4 students per workstation)
Enzyme, cellobiase, 1 ml
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 (DPTPs)
1.5 ml microcentrifuge tubes
15 ml conical tubes
1.5 ml standard disposable polystyrene cuvettes
Curriculum, including Teacher's Guide and Student Manual
Student Instruction Manual, available for download




Use the filters below to refine results!

Enzyme kinetics kit, includes enzyme, substrate, standard, solution, buffers, disposable pipets, tubes, and cuvettes for 32 students; inquiry-driven curriculum; education use only



Consumables refill reagents for use with Biofuel Enzyme kit (1665035EDU) and Biofuel Enzyme Reactions Kit (17001235EDU), contains enzyme, substrate, standard, 2x stop solution, 10x resuspension buffer, extraction buffer; education use only.



Number Description Options
Biofuel Enzyme Reactions Kit for AP Biology Student Manual, Rev A
Biofuel Enzyme Reactions Kit for AP Biology Teacher Manual Preview,Ver A
Biofuel Enzyme Reactions Kit for AP Biology Teacher Model Process,Rev A
AP Biology Alignment, ThINQ!™ Biofuel Enzyme Reactions Kit for AP Biology
Biofuel Enzyme Reactions Kit for AP Biology Planning Guide
Biofuel Enzyme Reactions Kit for AP Biology Quick Guide, Ver A
Experimental Design and Planning Worksheet
Mushroom Habitat Flashcards, Rev A
Biofuel Enzyme Reactions Kit Science Case Study, Ver A
Map of AP-Aligned Bio-Rad Kits with Learning Objectives
Science, Food, and Environment AP Strand Poster, Ver A