Ekstrak Kasar Isolat Bacillus sp Sebagai Agen Peluruh Biofilm
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Abstract
Abstract–Biofilms play an important role in the distribution and pathogenicity of bacteria. In our previous research, we demonstrated that our isolated Bacillus sp. (collection of Microbiology Lab, Universitas Surabaya) exhibited in vitro antibiofilm activity. Bacilli generally produce cellulase, which is one of the enzymes responsible for bacterial biofilm degradation. Based on this fact, this study aimed (1) to determine whether or not the crude extract of our Bacillus sp. isolate is able to degrade biofilms formed by Escherichia coli, Staphylococcus aureus, and water-crane bacteria, and (2) to determine whether such crude extract contains active cellulase. The research steps began with the preparation of Bacillus sp. crude extract, biofilm degradation assay, cellulase activity assay on CMC Agar, and cellulase detection by SDS-PAGE and electrophoresis zymogram. The crude extract of Bacillus sp. effectively degraded the biofilm 30 min after contact. The crude extract decreased biofilms of Escherichia coli 23,56%, Staphylococcus aureus 27,78%, and water-crane bacteria 37,98%. Based on the zymogram result, it was shown that the crude extract contained active cellulase, with a size of approximately 35 kDA. It was concluded that the crude extract of Bacillus sp. has potential as an anti-biofilm agent and exhibits active cellulase activity.
Keywords: Bacillus sp., crude extract, biofilm, SDS-PAGE, zymogram
Abstrak–Biofilm mempunyai peran penting dalam penyebaran dan sifat patogenitas bakteri patogen. Pada penelitian sebelumnya telah dibuktikan bahwa isolat Bacillus sp. (koleksi Laboratorium Mikrobiologi, Universitas Surabaya) mempunyai aktivitas anti-biofilm secara in vitro. Bacillus umumnya menghasilkan selulase sedangkan selulase diketahui dapat meluruhkan biofilm bakteri. Berdasarkan hal di atas, penelitian ini bertujuan (1) untuk mengetahui apakah ekstrak kasar isolat Bacillus sp. ini dapat meluruhkan biofilm Escherichia coli dan Staphylococcus aureus, dan bakteri air keran; dan (2) untuk mengetahui apakah dalam ekstrak kasar terdapat selulase. Metode penelitian ini meliputi penyiapan ekstrak kasar sel Bacillus sp., pengujian aktifitas meluruhkan biofilm, uji aktivitas selulase pada Agar CMC, dan analisis keberadaan aktivitas selulase pada ekstrak kasar Bacillus sp. menggunakan SDS-PAGE dan elektroforesis zymogram. Hasil penelitian menunjukkan bahwa ektrak kasar isolat Bacillus sp. bekerja paling baik pada perlakuan 30 menit setelah kontak dengan biofilm bakteri. Pemberian ekstrak kasar dapat menurunkan biofilm Escherichia coli sebesar 23,56%; dan biofilm bakteri Staphylococcus aureus sebesar 27,78%, dan biofilm bakteri air keran sebesar 37,98%. Berdasarkan hasil zymogram, telihat bahwa ekstrak kasar isolat Bacillus sp. memiliki selulase aktif dengan ukuran sekitar 35 kDa. Dapat disimpulkan bahwa ekstrak kasar isolat Bacillus sp. berpotensi sebagai agen peluruh biofilm dan memiliki selulase aktif.
Kata kunci: Bacillus sp., ekstrak kasar, biofilm, SDS-PAGE, zymogram
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