Biochemistry Practical Report Enzyme Kinetics

The kinetic effect of enzyme and substrate concentrations on rates of reaction.

Introduction

Why and how was this study performed?

Scientific purposes and objectives

Background information – general theory (notes & books)

Methods & Material

Refer to biochem prac notes

General view of the experimental procedures

Tabulate all materials and reagents used WITH concentrations of reagents and makes of apparatus

Table 1: Effect of enzyme concentration on the reaction rate.

Tube | mL 0.30mg/mL Substrate | mL 0.10 M Buffer | mL 0.15M NaCl | mL 200µg /mL Lysozyme |

1 | 1.90 | 2.10 | 2.00 | 0.00 |

2 | 1.90 | 2.05 | 2.00 | 0.05 |

3 | 1.90 | 2.00 | 2.00 | 0.10 |

4 | 1.90 | 1.95 | 2.00 | 0.15 |

5 | 1.90 | 1.90 | 2.00 | 0.20 |

Table 2: Effect of substrate concentration on the reaction rate.

Tube | mL 0.30mg/mL Substrate | mL 0.10 M Buffer | mL 0.15M NaCl | mL 200 µg /mL Lysozyme |

1 | 0.00 | 3.80 | 2.00 | 0.20 |

2 | 0.20 | 3.60 | 2.00 | 0.20 |

3 | 0.30 | 3.50 | 2.00 | 0.20 |

4 | 0.40 | 3.40 | 2.00 | 0.20 |

5 | 0.90 | 2.90 | 2.00 | 0.20 |

6 | 1.30 | 2.50 | 2.00 | 0.20 |

7 | 1.90 | 1.90 | 2.00 | 0.20 |

8 | 3.80 | 0.00 | 2.00 | 0.20 |

Table 3:Experiment 1—Preparation of buffer solution Experiment 2—Preparation of NaCl solution

Reagent formula | K2HPO4 | KH2PO4 |

Mass (g) | 0.674 | 0.834 |

Mass NaCl (g) |

0.438 |

Results

Table 4: Absorbance values at different enzyme concentrations. (Experiment 3)

| Absorbance at 450nm: |

Tube | 0sec | 15sec | 30sec | 45sec | 60sec |

1 | 0.298 | 0.298 | 0.298 | 0.298 | 0.298 |

2 | 0.292 | 0.283 | 0.277 | 0.270 | 0.264 |

3 | 0.270 | 0.265 | 0.252 | 0.239 | 0.230 |

4 | 0.288 | 0.262 | 0.241 | 0.225 | 0.216 |

5 | 0.292 | 0.264 | 0.242 | 0.223 | 0.214 |

Table 5: Absorbance values at different substrate concentrations. (Experiment 4)

| Absorbance at 450nm: |

Tube | 0sec | 15sec | 30sec | 45sec | 60sec |

1 | 0.005 | 0.005 | 0.005 | 0.005...