Pogostemon cablin Benth. var ‘Sidikalang’ leaves are the main source of patchouli oil which is used in the cosmetics and perfume industry because of its better fixative nature and provides the foreign exchange of around 60 % of Indonesia’s essential oil export revenues and is the largest patchouli oil supplier in the world with a contribution of 90 %. But apparently, several studies found that patchouli oil also has pharmacological activity such as aromatherapy, anti-influenza, antioxidants, and anti-microbial. This proves that leaves patchouli oil can be used as alternative medicine. Patchouli oil is composed of 20 main constituents, but the most important role as a determinant of the quality of patchouli oil is sesquiterpene patchouli alcohol (PA). The right extraction method is the most important factor so that the quality of patchouli oil is well maintained during the extraction process. Some methods that have been used for patchouli oil extraction include soxhlet, hydrodistillation, microwave-assisted hydrodistillation, microwave–air hydrodistillation, and water–bubble distillation. Microwave-assisted hydrodistillation and water–bubble distillation methods are more efficient methods for patchouli oil extraction. The microwave-assisted hydrodistillation method produces a yield of 1.9437 to 2.7227 % with a PA content of 26.32 % within 120 min to 126 min, and the water–bubble distillation method produces a yield of 2.40 % with a PA content of 61.53 % within 360 min.
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
Burkholderia cenocepacia H111 is an opportunistic pathogen which is difficult to eradicate due to its virulence factors, specifically biofilm. The ability of B. cenocepacia to form biofilm is regulated by quorum sensing (QS) system using a signal molecule. Environtmental bacteria often showed the ability to inhibit the QS of other species because they have special and unique metabolites or enzymes. In previous research several environtmental bacteria was isolated from few places in Java Island. However, until now the potential of these bacteria have not been investigated yet. This study was conducted with the aim to investigate the effect of these environmental bacterial isolates, i.e. Bacillus sp., Stenotrophomonas maltophilia, Enterobacter ludwigii and Kosakonia radicincitans on quorum sensing system of B. cenocepacia H111, particularly on quantities of acyl homoserine lactones (AHLs) and biofilm that were produced. The quantity of AHL molecule is measured by biosensor assay using Escherichia coli JM109 pSB401 while the quantity of biofilm formation was quantified by crystal violet method. The result showed that supernatants of all these environtmental bacteria have no impact in decreasing of AHL nor biofilm. On the other hands, all the crude extracts showed a reduction either to AHL molecule and biofilm formation. The Bacillus sp. crude extract revealed the most reducing effect, with 84 % for AHL molecule and 71 % for biofilm. It can be concluded that crude extracts of all environmental bacteria have quorum quenching activity to B. cenocepacia H111 cells, specifically in reducing AHL levels and biofilm quantity. Crude extract from Bacillus sp. gave the highest inhibitory effect.
Isolat 22 adalah kode isolat bakteri resisten logam Merkuri (Hg) yang didapatkan pada sampel air limbah pertambangan logam (tailing). Isolat 22 adalah isolat yang paling berdaya tahan tinggi terhadap Hg, dari total 24 isolat yang berhasil diisolasi. Pada paper ini akan dilaporkan hasil identifikasi spesies bakteri isolat 22 menggunakan teknik PCR - sekuensing gen 16S rRNA dan karakter pertumbuhan dan resistensi isolat 22 terhadap Hg. Dari hasil sekuensing 16S rRNA, diketahui bahwa isolat 22 sangat dekat kekerabatannya dengan spesies Stenotrophomonas maltophilia (identity 99 % dan query covered 90 %). Isolat 22 tumbuh dengan baik pada medium NB-Hg dengan kadar Hg 50 dan 100 ppm. Semakin tinggi kadar Hg dalam medum, semakin lama fase lag, walaupun jumlah sel pada medium di fase awal sama.
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