Isolation and Identification of Indigenous Lactic Acid Bacteria by Sequencing the 16S rRNA from Dangke, A Traditional Cheese from Enrekang, South Sulawesi
DOI:
https://doi.org/10.3923/pjn.2017.384.392Keywords:
16S rRNA, dangke, identification, isolation, lactic acid bacteriaAbstract
Background and Objective: Dangke is a traditional cheese from South Sulawesi that has been developed by the people of the Enrekang district throughout history. The microbiota of this cheese consists of a wide variety of bacterial species. The majority of which belongs to Lactic Acid Bacteria (LAB) genera. The indigenous LAB of dangke could be a potential source of starter cultures and probiotics. The aim of this study was to isolate the LAB from dangke and identify them by 16S rRNA sequencing. Methodology: The dangke from Enrekang, South Sulawesi were collected. The LAB were identified by morphology (Gram staining and cell form), physiology (growth and viability in 6.5% NaCl and temperatures of 15, 37 and 45°C), biochemistry (catalase-negative test and CO2 production) and survival at low pH (2, 3, 4 and 7.2) and in bile salts (0.3%). Results: The results showed that 30 isolates were identified as LAB with Gram-positive, catalase-negative and rod-shaped characteristics. Ten LAB isolates from dangke had highest tolerance to low pH and bile salts. The isolates that were resistant to low pH and bile salts were A123K, A113L, A323L, B111K, B212K, B221L, B312K, B323K, C113L and C222L. The 16S rRNA gene could be amplified by Polymerase Chain Reaction (PCR) from 5 isolates (A323L, B111K, B323K, C113L and C222L) to obtain a single band on a 1% agarose gel. Conclusion: Identification by 16S rRNA gene sequencing showed all isolates were identified as Lactobacillus fermentum with a similarity index of approximately 99-100%.
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