Isolation, Identification and Phylogenetic Analysis of Some Bacteriocins Producing Paenibacillus Species from Sheep Milk and Locally Fermented Alcoholic Beverage

Samples of sheep milk and locally fermented alcoholic beverage (burukutu) were investigated for the presence of bacteriocin producing Paenibacillus species. Serial dilutions were made and the appropriate dilutions inoculated on MRS agar using spread plate method. Gram positive rods bacteria were isolated from the samples and identified as Paenibacillus yanchengensis from burukutu sample and Paenibacillus yonginensis from sheep milk sample. The cell free extracts from Paenibacillus yanchengensis grown in a broth had antimicrobial activity against Listeria monocytogenes, Staphylococcus aureus and Salmonella sp. but inactive against E. coli while cell free extract from Paenibacillus yonginensis grown in MRS broth had antimicrobial activity against Listeria monocytogenes and Staphylococcus aureus but inactive against Salmonella sp. and E. coli. The bacteria universal primer (27F, 1492R) for 16S rRNA gene was used to confirm the bacteria status of these isolates and the PCR products of 16S rRNA gene primer was sequenced using Sanger method. The BLAST analysis of the sequences also identified the two isolates as Paenibacillus yanchengensis from burukutu sample and Paenibacillus yonginensis from sheep milk sample. Consequently, the BLAST result was used to construct the phylogenetic trees for the two Paenibacillus species and phylogenetic analysis.