Isolation and Identification of Lactic Acid Bacteria from Spontaneously Fermented Kunun-zaki using RAPD-PCR Analysis
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Abstract
Sixty lactic acid bacteria (LAB) were isolated from eighteen types of kunun-zaki produced naturally using millet, sorghum and maize each singly or in combinations with malted grains (millet, sorghum, maize, wheat and paddy rice) and characterized using conventional methods (biochemical and cultural) and molecular analyses. Seven morphotypes (Mt1-Mt7) were revealed among the 60 LAB isolates. Twenty LAB were selected based on the differences in their biochemical and physiological properties and these were subjected to DNA isolation and purification. The Agarose gel electrophoresis of the twenty (20) extracted DNA revealed very pure DNA electrophoretic pattern. The concentration of the DNA sample ranged between 0.7 mg/ml to 4.0 mg/ml and the purity index ranged from 1.5 to 1.9; indicating they were all very pure and can easily be used in Polymerase chain reaction (PCR) analysis. A highly polymorphic primer, OPB20 primer, was selected and used to analyze the 20 LAB DNA of which seven (7) isolates were identified to be Lactobacillus brevis, twelve (12) isolates were Lactobacillus plantarum and one isolate was identified as Lactobacillus acidophilus and these organisms could be potentially useful industrially.
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