EGUNJOBI, ADEYINKA KAMORU and Covenant University, Theses (2017) TANNASE PRODUCTION BY BACTERIAL ISOLATES FROM DETERIORATED BAMBARA (Vigna subterranean (L) Verde) NUTS. Masters thesis, COVENANT UNIVERSITY.
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Abstract
Tannase are hydrolytic enzymes that can be obtained from fungal and bacterial sources. They are used in the preparation of wine, beer, instant tea, coffee flavoured soft drinks and for clarification of juice. They are also used for treatment of waste water contaminated with polyphenolic compounds such as tannins which has been associated with environmental pollution. The immense potential of tannase has made it very important to optimize the production process to achieve maximum yields. Thirty-five grammes of Bambara nuts were thoroughly sorted and cleaned. Serial dilutions and subculturing of samples was carried out to obtain pure bacteria isolates. The isolates were screened for tannase production. Tannase producing bacterial isolates were identified based on clear zones of hydrolysis on tannin supplemented agar plates. They were further characterized by subjecting the isolates to various biochemical and molecular tests. The activity of the tannase was determined after termination using bovine serum albumin solution. Partial purification was carried out by ammonium sulphate precipitation. The optimum conditions of the tannase were determined using three parameters of temperature, pH and substrate concentration. Seven bacterial isolates (Six Gram positive and One Gram negative) were obtained. Two of the bacterial isolates (Isolate code G and C) were tannase producing bacteria with the highest clear zones of hydrolysis. Optimum conditions were ascertained at pH 6.0, temperature of 37oC and substrate concentration of 1.0% for isolate G while isolate code C had pH 6.0, temperature of 35oC and substrate concentration of 1.2% (w/v). Molecular identification of the tannase producing bacterial isolates revealed isolate code G as Bacillus pumilus strain AJ-2 (Accession number MF083685) and isolate code C has having 94% similarity with Aeromonas sanarellii strain A2-67. The Michaelis-mentens constant (Km) which is also known as the dissociation constant is the substrate concentration at half maximum velocity. Calculated from the Lineweaver-Burk plot, the apparent Km for the hydrolysis of tannic acid by tannase producing bacteria isolate code C and G were approximately 0.18mg/ml and 0.06mg/ml respectively. The results of this study established that Bambara nuts are host to tannase producing bacteria. This is the first report of tannase from the strains of bacterial isolates studied.
| Item Type: | Thesis (Masters) |
|---|---|
| Subjects: | Q Science > Q Science (General) Q Science > QH Natural history > QH301 Biology |
| Divisions: | Faculty of Medicine, Health and Life Sciences > School of Biological Sciences |
| Depositing User: | Mrs Hannah Akinwumi |
| Date Deposited: | 02 Jun 2020 10:41 |
| Last Modified: | 02 Jun 2020 10:41 |
| URI: | http://eprints.covenantuniversity.edu.ng/id/eprint/13351 |
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