Sains Malaysiana 49(9)(2020): 2129-2139

http://dx.doi.org/10.17576/jsm-2020-4909-11

Optimization of Enrichment and Isolation Media of Thermophilic Hydrocarbonoclastic Bacteria Isolated from Oil Reservoir of West Java for Microbial Enhanced Oil Recovery (MEOR) Application

(Pengoptimuman Media Isolasi dan Pengkayaan Bakteria Hidrokarbonoklastik Termofili yang Dipencilkan daripada Takungan Minyak Jawa Barat untuk Aplikasi Perolehan Kembali Minyak Galakan Mikrob (MEOR))

DEA INDRIANI ASTUTI, ISTY ADHITYA PURWASENA*, GREGORIUS GILANG SATRIO, KELVIN RACHMAD ANDIKA, GHAIDA ZAINIYA MILLATI, TERA WAHYU DWININGRUM & EVITA MERIANI

Department of Microbiology, School of Life Science and Technolog, Bandung Institute of Technology, Jalan Ganesha 10, Bandung West Java, Indonesia

Received: 15 January 2020/Accepted: 10 May 2020

ABSTRACT

Microbial enhanced oil recovery (MEOR) is a tertiary oil recovery process which aims to increase the crude oil recovery uptake to 55% from its original oil in place. In its application, indigenous microorganisms are commonly used as the main biological agents, but the isolation processes become more complicated due to environment’s extreme condition. This study aims to determine the optimum formulation for the enrichment and isolation medium, in order to obtain a highly diverse of indigenous hydrocarbonoclastic bacteria. The isolation process was carried out in two stages; the isolation of mild oil fraction degrading bacteria, and isolation of the heavy oil fraction degrading bacteria which confirmed by the SARA (saturated, aromatic, resinic, and asphaltic) fraction chromatography analysis. Four enrichment and five isolation media were formulated based on the distinction in the macro-and micronutrient composition. Overall, a total of 92 isolates were successfully isolated. The highest bacterial diversities were achieved in the enrichment medium B (crude oil used as carbon source which supplemented with sodium lactate) and isolation medium I (nutrient agar with twice agar concentration), each with 37 and 33% of obtained isolates, respectively. The newly formulated medium can be further utilized to isolate various types of indigenous bacteria from the oil well which were able to degrade SARA fraction from crude oil. The result showed that ABG2-7 as the isolate with the highest activity on SARA degradation. The amount of 15% inoculum of ABG2-7 isolate was able to degrade heavy crude oil, thus might serve as potential bacteria for MEOR application.

Keywords: Bacterial diversities; enrichment media; isolation media; MEOR

ABSTRAK

Perolehan kembali minyak galakan mikrob (MEOR) merupakan proses perolehan kembali minyak tertier yang bertujuan untuk meningkatkan perolehan kembali minyak mentah sehingga 55%. Mikroorganisma asli umumnya digunakan sebagai agen biologi tetapi proses pengasingan mikroorganisma tidak mudah untuk dilakukan kerana keadaan persekitaran yang ekstrim. Penyelidikan ini bertujuan untuk menentukan pengoptimuman media pengasingan dan pengkayaan bakteria bagi mendapatkan bakteria hidrokarbonoklastik yang asli. Proses pengasingan dilakukan dalam 2 tahap iaitu pengasingan bakteria yang mendegradasi pecahan minyak secara berat atau ringan dan disahkan dengan melakukan analisis kromatografi SARA (ketepuan, aromatik, resin dan asfaltena). Sebanyak 4 media pengkayaan dan 5 media pengasingan diformulasikan berdasarkan komposisi pembezaan pada mikro dan makro nutrien. Hasilnya, sebanyak 92 sampel pencilan berjaya diasingkan. Sampel pencilan tertinggi adalah pada sampel media B (minyak mentah yang mempunyai natrium laktat digunakan sebagai sumber karbon) dan media I (agar bernutrien dengan kepekatan 2 kali ganda) masing-masing dengan 37 dan 33% pencilan. Melalui medium ini juga, terdapat pelbagai jenis bakteria asli daripada takungan minyak boleh diasingkan kerana bakteria ini mampu mendegradasikan SARA daripada minyak mentah. Hasil menunjukkan sampel pencilan ABG2-7 memiliki kemampuan yang terbaik dalam mendegradasi SARA. Jumlah inokulum paling optimum daripada pencilan ABG2-7 yang digunakan untuk mendegradasi pecahan berat minyak mentah adalah sebanyak 15%, sekaligus berpotensi sebagai bakteria yang boleh digunakan untuk aplikasi MEOR.

Kata kunci: Kepelbagaian bakteria; media perkayaan; media pengasingan; MEOR

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*Corresponding author; email: isty@sith.itb.ac.id