Analysis Of Bacteria Community In The Plastic Waste

Dhebby Purba; Veibe Warouw; Rizald Rompas; Deiske Sumilat; Reni Kreckhoff; Elvy Ginting

doi:10.35800/jip.8.2.2020.29877

Analysis Of Bacteria Community In The Plastic Waste

Dhebby Purba, Veibe Warouw, Rizald Rompas, Deiske Sumilat, Reni Kreckhoff, Elvy Ginting

Abstract

Plastic waste is a general problem for the environment. Therefore, more ways of plastic waste management are required. One of the plastic waste management process is microbiological biodegradation using bacteria. This research aims to identify the bacterial community in Polyethylene Terephthalate type plastic waste by molecular analysis. In order to identify the community of bacteria, it is necessary to isolate the DNA of uncultured bacteria. In this research, the isolation of the DNA of uncultured bacteria from plastic waste was carried out by the Qiagen DNeasy® PowerSoil Pro Kit handbook. The DNA genome was subsequently examined using electrophoresis gel and UV-Vis spectrophotometer. The DNA genome was then amplified using Polymerase Chain Reaction and the bacteria were identified using Next Generation Sequencing (NGS) analysis. On the results of this study, the uncultured bacteria DNA from plastic waste was isolated with purification value of 1. The amplified DNA genome was detected by DNA bands through gel electrophoresis gel at ~1500 bp. The bacteria in the plastic waste were Proteobacteria, Cyanobacteria, Actinobacteria, Chlorobi, Bacteroidetes, Firmicutes, Chloroflexi, Thaumarchaeuta and Gemmattimonadetes with the highest relative abundance of Proteobacteria.

Keywords : Bacteria, Deoxyribo Nucleic Acid ( DNA), Plastic Waste, Uncultured

Abstrak

Sampah plastik merupakan masalah bagi lingkungan oleh sebab itu diperlukan pengelolaan yang baik. Salah satu pengelolaan sampah plastik yakni dengan cara biodegradasi secara mikrobiologi menggunakan bakteri. Penelitian ini bertujuan untuk mengetahui komunitas bakteri pada sampah plastik, khususnya jenis Polyethylene Terephthalate secara molekuler. Untuk analisis bakteri secara molekuker, diperlukan tahapan isolasi DNA bakteri. Pada penelitian ini isolasi DNA bakteri tanpa kultivasi dari sampah plastik dilakukan dengan mengikuti prosedur ekstraksi DNeasy® PowerSoil Pro Kit, Qiagen dengan menggunakan proses pengikisan pada kedua permukaan plastik sebagai tahapan preparasi sampel. Hasil ekstraksi DNA selanjutnya diuji menggunakan elektroforesis gel dan spektofotometer UV-Vis. Genom DNA kemudian diamplifikasi menggunakan Polymerase Chain Reaction dan bakteri diidentifikasi dengan Next Generation Sequencing analysis. Hasil penelitian menujukkan bahwa, DNA bakteri tanpa kultivasi dari sampah plastik berhasil diisolasi dan diamplifikasi yang ditunjukkan dengan terbentuknya pita DNA lewat hasil elektroforesis gel pada panjang 1500 bp. Bakteri yang terdapat pada plastik jenis PET adalah Proteobacteria, Cyanobacteria, Actinobacteria, Chlorobi, Bacteroidetes, Firmicutes, Chloroflexi, Thaumarchaeuta dan Gemmattimonadetes dengan kelimpahan relatif tertinggi adalah Proteobacteria

Kata kunci: Bakteri, Deoxyribo Nucleic Acid ( DNA), Sampah Plastik, Tanpa Kultivasi

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DOI: https://doi.org/10.35800/jip.8.2.2020.29877