Keanekaragaman Molekuler Dengan Pendekatan Filogenetik Pada Isolat Bakteri Resisten Kromium dari Lahan Eco Urban Farming

Sari, Adisya Prima Nurmalita (2018) Keanekaragaman Molekuler Dengan Pendekatan Filogenetik Pada Isolat Bakteri Resisten Kromium dari Lahan Eco Urban Farming. Masters thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Salah satu logam berat yang banyak digunakan di industri adalah kromium (Cr), kromium heksavalen merupakan agen oksidator kuat dan sangat mudah larut dalam air sehingga bersifat toksik. Beberapa isolat bakteri yang diisolasi dari lahan Eco Urban Farming ITS resisten terhadap kromium sampai dengan 200 mg/L, yaitu isolat A1a, A3, A5, A7 dan A9. Bakteri resisten terhaap Cr dapat melalui mekanisme bioadsorpsi atau membentuk protein transporter (ChrA) dan protein regulator (ChrB) yang dikode oleh gen chrA dan chrB. Penelitian ini bertujuan untuk mengetahui keanekaragaman molekuler bakteri resisten kromium dari lahan eco urban farming sehingga dapat diidentifikasi strain atau spesiesnya. Uji resistensi terhadap kromium dilakukan dengan isolat ditumbuhkan pada media NA-K2Cr2O7. Viabilitas isolat terhadap kromium dilakukan melalui pengukuran optical density menggunakan spektrofotometer UV-Vis dan divisualisasikan dengan kurva pertumbuhan. Bioakumulasi kromium diukur menggunakan Atomic Absorption Spectroscophy (AAS). Identifikasi spesies menggunakan marka gen 16S rRNA. Profil protein menggunakan SDS-PAGE. Deteksi gen chrA dan chrB menggunakan primer spesifik. Semua isolat mampu tumbuh dengan baik pada media NA-K2Cr2O7 hingga konsentrasi ≤250 mg/L. Pola pertumbuhan isolat pada media yang mengandung kromium relatif lebih rendah dibandingan kontrol. Isolat A9 memiliki efisiensi bioakumulasi tertinggi, yaitu 97% dari 10 mg/L kromium yang dipaparkan, sedangkan isolat A1a memiliki efisiensi bioakumulasi terendah, yaitu 64.6% dari 30 mg/L kromium yang dipaparkan. Identifikasi spesies dengan marka gen 16S rRNA menunjukkan isolat A1a adalah Bacillus aerius, isolat A3 adalah Enterobacter tabaci, isolat A5, A7 dan A9 adalah Bacillus cereus. Kemotaksonomi dengan pendekatan profil protein menunjukkan hasil yang sama dengan rekonstruksi pohon filogenetik berdasarkan marka gen 16S rRNA. Gen chrA dan chrB tidak terdeteksi pada semua isolat. ============================================================ Chromium (Cr) is one of heavy metal widely used in industries. Hexavalent chromium is a strong oxidizing agent and easy to dissolved in the water so that it is toxic to living things. Some of bacteria isolated from Eco Urban Farming of ITS Surabaya are resistant to chromium until 200 mg/L, isolate A1a, A3, A5, A7 dan A9. Bacterial resistance to chromium could be through bioadsoprtion mechanism or through transporter protein (ChrA) and regulator protein (ChrB) which encoded by chrA and chrB gene, respectively. The aim of this research is to obtain the molecular diversity of chromium-resistant bacteria in order to identify strain or species. Resistance assay was done by growing the isolates on the NA-K2Cr2O7 media. The viability of isolates to chromium was done by measuring optical density using spectrophotometer UV-Vis and visualized by growth curve. Chromium bioaccumulation was measured by Atomic Absorption Spectroscopy (AAS). Identification of species was done using 16S rRNA gene marker. Protein profile was done by SDS-PAGE electrophoresis. The presence of chrA and chrB gene was confirmed using specific primers. Each isolate grew well on NA-K2Cr2O7 media until ≤250 mg/L. Growth pattern of isolates on media containing chromium was relatively lower than control. Isolate A9 has highest bioaccumulation efficiency, which could accumulate 97% of 10 mg/L chromium exposed. Isolate A1a has the lowest bioaccumulation efficiency, which could accumulate 64.6% of 30 mg/L chromium exposed. Identification based on 16S rRNA gene marker showed that isolate A1a is Bacillus aerius, isolate A3 is Enterobacter tabaci, isolate A5, A7, and A9 is Bacillus cereus. Chemotaxonomy based on protein profile using SDS-PAGE showed the same result with phylogenetic tree construction based on 16S rRNA gene marker. chrA and chrB gene were not detected in all isolates.

Item Type: Thesis (Masters)
Uncontrolled Keywords: 16S rRNA, bioakumulasi, kromium, profil protein, resistensi
Subjects: Q Science > Q Science (General)
Q Science > QP Physiology > QP624 Molecular biology.
Divisions: Faculty of Mathematics and Science > Biology
Depositing User: Sari Adisya Prima Nurmalita
Date Deposited: 02 May 2018 03:24
Last Modified: 03 May 2018 08:53
URI: http://repository.its.ac.id/id/eprint/51137

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