- Title
- In silico analysis, isolation and kinetic characterisation of red algae (Rhodophyta) catalases
- Creator
- Nodangala, Sinovuyo
- Subject
- Red algae Marine algae
- Date Issued
- 2018
- Date
- 2018
- Type
- Thesis
- Type
- Masters
- Type
- MSc
- Identifier
- http://hdl.handle.net/10353/14626
- Identifier
- vital:40024
- Description
- Catalase (EC: 1.11.1.6) is produced by almost all aerobic organism ranging from bacteria to human and is an essential antioxidant enzyme that catalyses the conversion of hydrogen peroxide to water and molecular oxygen, therefore neutralising hydrogen peroxide toxicity. The present study aimed to purify and characterise catalase from Pachymenia orbitosa, a marine red algae found in the Algoa Bay region of South Africa. Bioinformatics analysis was performed to confirm the presence of a catalase gene in the red algae genome. In silico analysis of the Chondrus crispus genome was performed to predict the functional and structural characterisation of the protein encoded by the gene. The Pachymenia orbitosa catalase enzyme was purified to homogeneity using 60% ammonium sulphate precipitation and Sephacryl S-200 gel filtration chromatography. InterProScan confirmed that the Chondrus crispus genome encodes a catalase protein, which is from the mono-functional and heme-dependent catalase-like superfamily. The purified Pachymenia orbitosa catalase enzyme displayed a specific activity of 23 000 units per mg of protein with a 14.94% recovery and 222.91 fold purification. Sodium dodecyl sulphatepolyacrylamide gel electrophoresis indicated a single band, with a molecular weight of around 50kDa. The catalase enzyme exhibited a maximum activity at pH 7 and a temperature of 30℃. It was stable up to 40℃ and rapidly denatured at temperatures above this. The Km and Vmax values for the purified catalase, using hydrogen peroxide as a substrate, were determined from the Lineweaver-Burk plot to be 22.22mM and 1.11x10-4mM.min-1 , respectively, while from the Hanes-Woolf plot, to be 23.4mM and 1.17x10-4mM.min-1 , respectively. The Heme catalase inhibitor (ferricyanide) inhibited the enzyme activity markedly, while sodium chloride and citric acid had only a slight inhibitory effect. Copper sulphate showed a slight stimulatory effect. The physiochemical properties suggest a good application potential in both the pharmaceutical and food industries
- Format
- 105 leaves
- Format
- Publisher
- University of Fort Hare
- Publisher
- Faculty of Science and Agriculture
- Language
- English
- Rights
- University of Fort Hare
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UFH Department of Biochemistry & Microbiology
| Thumbnail | File | Description | Size | Format | |||
|---|---|---|---|---|---|---|---|
| View Details Download | SOURCE1 | NODANGALA Dissertation.pdf | 3 MB | Adobe Acrobat PDF | View Details Download |