Identifikasi Bakteri Pereduksi Logam Pb dalam Bioremediasi Sampel Air Sungai Citarum Menggunakan Analisis Gen 16s rRNA

Identification of Pb Metal Reducting Bacteria in Bioremediation from Citarum River Water Samples Using 16s rRNA Gene Analysis

Authors

DOI:

https://doi.org/10.33084/bjmlt.v5i2.5102

Keywords:

Bacillus cereus, Lead (Pb), 16S rRNA Gene Sequencing

Abstract

The bioremediation technique is an effort to remove the heavy metal Pb in samples of Citarum river water. High concentrations of heavy metals harm the environment, so Pb metal-reducing bacteria are needed as a pollution solution. Bioremediation uses bacteria. Bacteria are identified by molecular methods, namely sequencing analysis techniques, because they are more accurate. To determine the nucleotide sequence of Pb metal-reducing bacteria isolates and which bacterial species can reduce Pb metal. The methodology used is primary data collection, namely conducting research directly, namely the stages of bacterial purification through gram staining techniques, pure colonies, then proceeding to the DNA isolation stage. The isolated DNA is then electrophoresed. The electrophoresis results are then purified and proceed to the sequencing stage. The feasibility test results were to obtain pure DNA from the reducing bacteria. The next step was PCR amplification of the 16S rRNA gene of Pb-reducing bacteria. The research results started at the DNA isolation stage and obtained a purity of 1.80. The PCR results were in the form of amplification of the 16S rRNA gene of Pb-reducing bacteria. Electrophoresis obtained the results of DNA bands measuring ± 1400 bp—phylogenetic tree analysis with 99.60% relatedness to Bacillus cereus, which can reduce Pb. Bacillus cereus species can carry out detoxification mechanisms against the toxic effects of Pb metals, such as—biosorption, bioaccumulation, reduction, solubilization, precipitation (formation of extracellular complexes), and methylation.

Downloads

Download data is not yet available.

References

Ahmad, R.Z., 2018. Mycoremediation to remove heavy metal pollution in post-mining areas for farmland utilization.

Angraini, N., Agustina, T.E. and Hadiah, F., 2022. Pengaruh pH dalam Pengolahan Air Limbah Laboratorium Dengan Metode Adsorpsi untuk Penurunan Kadar Logam Berat Pb, Cu, dan Cd. Journal Ilmu Lingkungan, 20(2), pp.345-355.

Awad, G.E., Mostafa, H., Danial, E.N., Abdelwahed, N.A. and Awad, H.M., 2015. Enhanced production of thermostable lipase from Bacillus cereus ASSCRC-P1 in waste frying oil based medium using statistical experimental design. Journal of Applied Pharmaceutical Science, 5(9), pp.007-015.

Besser, J., Carleton, H.A., Gerner-Smidt, P., Lindsey, R.L. and Trees, E., 2018. Next-generation sequencing technologies and their application to the study and control of bacterial infections. Clinical microbiology and infection, 24(4), pp.335-341.

Besser, J., Carleton, H.A., Gerner-Smidt, P., Lindsey, R.L. and Trees, E., 2018. Next-generation sequencing technologies and their application to the study and control of bacterial infections. Clinical microbiology and infection, 24(4), pp.335-341.

Das, N. and Chandran, P., 2011. Microbial degradation of petroleum hydrocarbon contaminants: an overview. Biotechnology research international, 2011.

Dharmayanti, N.L.P.I., 2011. Filogenetika molekuler: metode taksonomi organisme berdasarkan sejarah evolusi. Wartazoa, 21(1), pp.1-10.

Gofar, N., 2012. Aplikasi Isolat Bakteri Hidrokarbonoklastik asal Rizosfer Mangrove pada Tanah Tercemar Minyak Bumi. Jurnal Lahan Suboptimal: Journal of Suboptimal Lands, 1(2).

Hamidah, M.N., Rianingsih, L. and Romadhon, R., 2019. Aktivitas antibakteri isolat bakteri asam laktat dari peda dengan jenis ikan berbeda terhadap E. coli dan S. aureus. Jurnal Ilmu dan Teknologi Perikanan, 1(2), pp.11-21.

Harahap, M.R., 2018. Elektroforesis: Analisis elektronika terhadap biokimia genetika. CIRCUIT: Jurnal Ilmiah Pendidikan Teknik Elektro, 2(1).

Ihsan, Y.N., Fellatami, K., Permana, R., Mulyani, Y. and Pribadi, T.D.K., 2020. ANALISIS BAKTERI PEREDUKSI KONSENTRASI LOGAM TIMBAL Pb (CH3COO) 2 MENGGUNAKAN GEN 16S Rrna. Jurnal Kelautan: Indonesian Journal of Marine Science and Technology, 13(2), pp.151-162.

Intarapanich, A., Kaewkamnerd, S., Shaw, P.J., Ukosakit, K., Tragoonrung, S. and Tongsima, S., 2015. Automatic DNA diagnosis for 1D gel electrophoresis images using bio-image processing technique. BMC genomics, 16(12), pp.1-11.

Irawan, B., 2013. Karsinologi Dengan Penjelasan Deskriptif dan Fungsional. Airlangga University Press.

Irmawati.2013 Perubahan keragaman genetik ikan kerapu tikus (cromileptes altivelis) generasi pertama pada stok hatchery.

Jiménez-Montenegro, L., Mendizabal, J.A., Alfonso, L. and Urrutia, O., 2022. DNA extraction procedures and validation parameters of a real-time PCR method to control milk containing only A2 β-casein. Food Control, 142, p.109259.

Joko, T., Kusumandari, N. and Hartono, S., 2011. Optimasi metode PCR untuk deteksi Pectobacterium carotovorum, penyebab penyakit busuk lunak anggrek. Jurnal Perlindungan Tanaman Indonesia, 17(2), pp.54-59.

Kepel, B., 2015. Fatimawali. Penentuan jenis dengan analisis gen 16SrRNA dan uji daya reduksi bakteri resisten merkuri yang diisolasi dari feses pasien dengan tambalan amalgam merkuri di Puskesmas Bahu Manado. Jurnal Kedokteran Yarsi, 23(1), pp.45-55.

Langden, S.S., Budiharjo, A. and Kusharyoto, W., 2017. TRANSFORMASI DAN KLONING PLASMID PJ804: 77539 PADA E. coli TOP’10. Jurnal Akademika Biologi, 6(1), pp.65-70.

Lokapirnasari, W.P., Dewi, A.R., Fathinah, A., Hidanah, S. and Harijani, N., 2017. Effect of probiotic supplementation on organic feed to alternative antibiotic growth promoter on production performance and economics analysis of quail. Veterinary world, 10(12), p.1508.

Lott, S.C., Wolfien, M., Riege, K., Bagnacani, A., Wolkenhauer, O., Hoffmann, S. and Hess, W.R., 2017. Customized workflow development and data modularization concepts for RNA-sequencing and metatranscriptome experiments. Journal of biotechnology, 261, pp.85-96.

Lubis, K., 2014. Cara pembuatan pohon filogeni. Jurnal Pengabdian Kepada Masyarakat, 20(75), pp.66-69.

Murtiyaningsih, H., 2017. Isolasi DNA genom dan identifikasi kekerabatan genetik nanas menggunakan RAPD (Random Amplified Polimorfic DNA). Agritrop: Jurnal Ilmu-Ilmu Pertanian (Journal of Agricultural Science), 15(1).

Muzzazinah, 2017. Metode filogenetik pada indigofera. Prosiding Seminar Nasional Pendidikan Biologi Dan Biologi, 25–40

Narita, V., Arum, A.L., Isnaeni, S. and Fawzya, N.Y., 2014. Analisis bioinformatika berbasis web untuk eksplorasi enzim kitosanase berdasarkan kemiripan sekuens. Jurnal Al-Azhar Indonesia Seri Sains dan Teknologi, 1(4), pp.197-203.

Nikunjkumar, B.D., Dhruvil. 2012. Molecular identification of bacteria using 16s rDNA sequencing. Gujarat University, Gujarat, India.

Nurhidayati, S., Faturrahman, F. and Ghazali, M., 2015. Deteksi bakteri patogen yang berasosiasi dengan Kappaphycus alvarezii (Doty) bergejala penyakit ice-ice. Jurnal Sains Teknologi & Lingkungan, 1(2).

Pelczar, M.J & Chan, E.C.S, 2019 Dasar-dasar mikrobiologi. UI Press, Jakarta.

Raven and Johson, 2018. Teknik Perancangan Primer Untuk Sekuen Gen Mdr-1 Varian 1199 Pada Sampel Buffy Coat Pasien Anak Dengan Lla .1: 105-111.

Sabbathini, G.C. and Pujiyanto, S., 2017. Isolasi dan identifikasi bakteri genus Sphingomonas dari daun padi (Oryza sativa) di area persawahan Cibinong. Jurnal Akademika Biologi, 6(1), pp.59-64.

Suhendrayatna. 2001. Bioremoval logam berat dengan menggunakan mikroorganisme: Suatu Kajian Kepustakaan. Makalah. Disampaikan pada Seminar Bioteknologi, Kagoshima University, Tokyo.

Varghese, N.P.P., Joy, 2014. Microbiology Laboratory Manual Book. Kerala Agricultural University. India.

Wicaksono ,N.B, 2013. Mesin PCR pada Instrumentasi Laboratorium Klinik.125-137

Wulandari, D. and Purwaningsih, D., 2019. Identifikasi dan karakterisasi bakteri amilolitik pada umbi Colocasia esculenta L. secara morfologi, biokimia, dan molekuler. Jurnal Bioteknologi dan Biosains Indonesia, 6(2), pp.247-258.

Yulaipi, S. and Aunurohim, A., 2013. Bioakumulasi logam berat timbal (Pb) dan hubungannya dengan laju pertumbuhan Ikan mujair (Oreochromis mossambicus). Jurnal Sains dan Seni ITS, 2(2), pp.E166-E170.

Zein MSA & Sulandari S, 2009. Investigasi Asal Usul Ayam Indonesia menggunakan Sekuens Hypervariable-1 D-loop DNA Mitokondria. Jurnal Veteriner.10(1):41 - 49.

Downloads

Published

2023-04-28

How to Cite

Murtafi’ah, N., & Aeni, S. R. N. (2023). Identifikasi Bakteri Pereduksi Logam Pb dalam Bioremediasi Sampel Air Sungai Citarum Menggunakan Analisis Gen 16s rRNA: Identification of Pb Metal Reducting Bacteria in Bioremediation from Citarum River Water Samples Using 16s rRNA Gene Analysis. Borneo Journal of Medical Laboratory Technology, 5(2), 303–315. https://doi.org/10.33084/bjmlt.v5i2.5102