Hydrogen peroxidase coursework

They do so by take downing the activation energy. Enzymes contain an active site where a substrate, in this instance, the H peroxide binds to it and interrupt into H2O and O.

Hydrogen peroxidase coursework

Nakajima, in Methods in Enzymology1 Introduction Peroxidases are some of the most common and ubiquitous enzymes. Many peroxidases contain an iron-porphyrin derivative heme in their active site, and they can catalyze oxidation of a wide variety of organic compounds using hydrogen peroxide H2O2.

Despite the benefits of peroxidases, their full-scale use is still restricted due to their facile inactivation in the presence of H2O2 under supraphysiological conditions. In addition, their poor thermal and environmental tolerance reduces the range of practical uses of peroxidases.

Many efforts have been devoted to overcoming these problems.

Materials and Methods

Random mutagenesis based on evolutionary engineering has also improved the stability and enzymatic activity of known peroxidases under catalytic conditions Cherry et al. To date, these biological approaches have been reasonably successful for obtaining peroxidases bearing desirable stability and activity.

This methodology allows successive molecular modifications leading to further development of desired properties. Despite this advantage, studies on such engineered proteins have been scarce, possibly due to the intrinsic fragility of the scaffold proteins examined.

In this study, we decided to exploit a heme-protein from a thermophile as a scaffold for protein engineering. Cytochrome c Cyt c from Thermus thermophilus HB8 is a small kDa heme-protein expressed in the periplasmic space Than et al.

The mutagenesis we performed on Cyt c consisted of two steps. First, an active site on the heme iron was introduced by replacing Met69 with alanine M69A and a general acid—base catalyst was deployed by replacing Val49 with aspartic acid V49D Fig.

This result demonstrated the advantage of using an intrinsically thermostable protein as a scaffold to produce artificial peroxidases capable of acting at high temperatures.

Effect of Substrate Concentration on the Rate of Activity of Catalase | Owlcation

However, the mutant showed insufficient persistence in catalytic activity not due to denaturation, but due to heme degradation during the reaction cycle.

The major active intermediate was an oxo-ferryl heme with a protein radical localized predominantly on Tyr45, which is located close to the heme group and is connected to a heme propionate group through a hydrogen bond. The active intermediate could be transformed into a precursor for heme degradation by reaction with H2O2.

In the following sections, we describe the processes of transforming Cyt c to a thermally tolerant peroxidase based on spectroscopic characterization and kinetic analyses of reaction intermediates. We propose that proteins from thermophiles are useful resources not only for exploring a protein of desired function but also for constructing thermally stable bio-based materials amenable to rational molecular design based on the chemistry of a target function.Hydrogen peroxide reacts with ferrous horseradish per- oxidase and converts it to oxyperoxidase in a sequence of two reactions.

The first is the reaction of ferrous peroxidase with HzOz to form Compound II; the second is the reaction of Compound II with HzOz to form oxyperoxidase. The Decomposition of Hydrogen Peroxide To learn about Avogadro’s Hypothesis and the Ideal Gas Law.

To learn about gas phase Stoichiometry.

Hydrogen peroxidase coursework

Of course, the pressure and temperature of the gas must simultaneously be determined as well. Hydrogen peroxide is a strong oxidizing agent used in aqueous solution as a ripening agent, bleach, and topical anti-infective.

Introduction

It is relatively unstable and solutions deteriorate over time unless stabilized by the addition of acetanilide or similar organic materials.

The approach utilizes hydrogen peroxide as the enzyme’s primary substrate, a hypothetically relevant substrate given its presence in physiological conditions. Kinetic analysis of the A2E degradation has revealed a first-order constant, k cat /K M, of min-1 µM-1, a value comparable to several substrates of hydrogen peroxidase.

The peroxidase breaks down hydrogen peroxide to form water and oxygen, which binds to the indicator guaiacol in this experiment to prevent the cell from denaturing. The substrate allows the enzyme to react by changing it to its highly specific active site.

Apr 30,  · Here is an example of A-level biology coursework on the effect of substrate concentration (hydrogen peroxide) on the rate of activity of the enzyme yunusemremert.coms: 5.

Hydrogen peroxidase coursework
Peroxidase - an overview | ScienceDirect Topics