- Title
- Production and molecular characterization of peroxidases from novel ligninolytic proteobacteria and bacillus strains
- Creator
- Falade, Ayodeji Osmund
- Subject
- Peroxidase Catalase
- Date
- 2018
- Type
- Thesis
- Type
- Doctoral
- Type
- PhD
- Identifier
- http://hdl.handle.net/10353/9702
- Identifier
- vital:34823
- Description
- Inadequate quantity is a major impediment to the industrial application of peroxidase and other industrial enzymes. Consequently, efforts are geared towards increasing peroxidase production by searching for new microbes with enhanced production capacity. In this study, three novel ligninolytic bacteria: Raoultella ornithinolytica OKOH-1 (KX640917), Ensifer adhaerens NWODO-2 (KX640918) and Bacillus sp. FALADE-1 (KX640922) were optimized for peroxidase production and their peroxidases characterized using molecular and biochemical approaches. Molecular analysis confirmed the presence of peroxidase genes in the three bacteria. BLAST result and phylogenetic analysis of the deduced amino acid sequences suggested that Raoultella ornithinolytica OKOH-1 peroxidase (RaoPrx) belongs to a DyP-type peroxidase family while peroxidases from Ensifer adhaerens NWODO-2 and Bacillus sp. FALADE-1 are catalase-peroxidases. The peroxidase genes are available in the GenBank with MF370527, MF374336 and MF407314 as respective accession numbers. Upon optimization, Raoultella ornithinolytica OKOH-1 exhibited the highest peroxidase production at pH 5, 35 oC and 150 rpm. Biochemical characterization showed that RaoPrx had a wide substrate specificity as it was able to oxidize all the tested substrates in this study (ABTS, veratryl alcohol, guaiacol and pyrogallol), except 2, 6-Dimethoxyphenol. However, highest activity by the enzyme was recorded with pyrogallol as substrate. The enzyme had an optimum activity at pH 6 and 50 oC and was very stable at high temperatures (50 oC – 70 oC). Its pH stability was over a pH range of 5.0 – 7.0. Moreover, RaoPrx activity was significantly enhanced by Ag+, Cu2+, Zn2+and Fe2+ while Ca2+, Mg2+, Ba2+, Al3+, Co2+, NaN3 and EDTA inhibited the activity of the enzyme. Nevertheless, RaoPrx exhibited a remarkable dye-decolourizing activity on congo red and melanin, indicating the biotechnological potential of the enzyme in dye decolourization and development of cosmetic agent. Generally, the results from this study suggest that ligninolytic bacteria hold a great potential for enhanced peroxidase production that could meet the increasing industrial demand.
- Format
- 250 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 | SOURCE1 | PhD (Biochemistry) FALADE - May 2018.pdf | 3 MB | Adobe Acrobat PDF | View Details |