Phylogenetic analysis of bacteria producing antimicrobial agent isolated from Kukup Sand Beach Ambarwati Ambarwati, Broto Santoso, Aan Sofyan
Universitas Muhammadiyah Surakarta
Abstract
Microorganisms are a source of producing secondary metabolites, including bacteria as producers of antimicrobial substances. The purpose of this study was to molecularly identify antimicrobial-producing bacteria isolated from the Kukup sand Beach, Indonesia. The steps in this study included: rejuvenation of 9 bacterial isolates, antimicrobial testing of bacterial isolates against the tested bacteria, DNA isolation of the 9 bacterial isolates, PCR, 16S rRNA gene sequencing, reconstruction and phylogenetic tree analysis of the 9 bacterial isolates. The results showed that among of 9 bacterial isolates, 8 isolates were able to inhibit minimum one of 4 test bacteria isolates with a diameter of inhibition area between 7 and 22.5 mm. Isolate ARA-2 was an isolate that was able to inhibit the growth of S. aureus ATCC 25923 with the greatest resistance (22.5 mm), while isolate BR2-9 had the weakest inhibition against B. subtilis FNCC 0060, with a diameter of inhibition zone of 7 mm. One isolate was able to inhibit 2 test bacteria, namely isolate BR1-27 which was able to inhibit B. subtilis FNCC 0060 with an inhibition area diameter of 17.5 mm and P. auroginosa ATTC 27853 with an inhibition area diameter of 7.5 mm. Based on the results of the phylogenetic tree analysis, it was known that isolate ARA-2 was a sister clade of Bacillus flexus strain IFO15715 with 100% similarity. Meanwhile, isolate BR2-9 was a sister clade of Bacillus cereus strain CCM 2010 with 100% similarity. And isolate BR1-27 has the closest kinship with Paenibacillus lautus strain JCM 9073 with a similarity of 82%.
Keywords: Streptomyces, Kukup Sand Beach, Antimicrobial, Phylogenetic Tree
