Octaviany, Evi and Irfan Mustafa,, S.Si., M.Si., Ph.D and Dr. Suharjono,, MS (2019) Efektivitas Kombinasi Bakteri Hidrokarbonoklastik Dengan Saponin Sapindus Rarak Dalam Menurunkan Kadar Total Petroleum Hidrokarbon. Magister thesis, Universitas Brawijaya.
Abstract
Surfaktan memiliki peran penting dalam meningkatkan biodegradasi minyak bumi. Salah satu surfaktan alami yang jarang dieksplorasi dalam membantu biodegradasi limbah minyak bumi adalah saponin. Tujuan penelitian ini untuk menganalisis pengaruh kombinasi saponin komersial dari tanaman Sapindus rarak dengan bakteri hidrokarbonoklastik terisolasi dalam menurunkan kadar total petroleum hidrokarbon. Bakteri hidrokarbonoklastik diisolasi dari sampel tanah tercemar minyak bumi di Desa Wonocolo, Kabupaten Bojonegoro, Provinsi Jawa Timur, dengan metode enrichment dalam medium Bushnell-Haas broth dengan konsentrasi minyak bumi 0,5 %. Isolat bakteri yang diperoleh diseleksi dengan dua tahap. Tahap pertama, isolat diinokulasikan dalam medium nutrient broth yang mengandung saponin 0 %, 8 %, dan 12 % untuk memperoleh isolat bakteri yang toleran terhadap saponin. Tahap kedua, isolat diinokulasikan ke dalam medium Bushnell-Haas broth yang mengandung saponin 0,5 % untuk memperoleh isolat bakteri yang tidak dapat mendegradasi saponin. Isolat yang lolos seleksi dibuat kurva pertumbuhannya dalam medium Bushnell-Haas broth dengan konsentrasi minyak bumi 0,5 % dan dipilih berdasarkan laju pertumbuhan tercepat. Isolat terpilih digunakan pada uji biodegradasi minyak bumi 0,5 % dalam medium Bushnell-Haas broth menggunakan Rancangan Acak Lengkap (RAL) faktorial dengan perlakuan keberadaan isolat terpilih dan saponin 0,5 %. Penurunan total petroleum hidrokarbon (TPH) minyak bumi dihitung berdasarkan kandungan TPH dengan metode gravimetri. Data TPH dianalisis ragam ANOVA (Analysis of Variance) pada taraf α 0,05. Bakteri unggul diidentifikasi berdasarkan sekuen 16S rDNA dan dikonstruksikan dalam bentuk pohon filogeni. Bakteri sebanyak 34 isolat telah diisolasi dan dimurnikan dari tanah tercemar minyak bumi. Hasil seleksi tahap pertama, mendapatkan sembilan isolat yaitu IHT1.3, IHT1.5, IHT3.8, IHT3.14, IHT3.17, IHT3.19, IHT3.20, IHT3.23, dan IHT3.24 yang memiliki toleransi tinggi terhadap saponin 12 %. Berdasarkan hasil seleksi tahap berikutnya, tiga dari sembilan isolat tersebut (IHT1.3, IHT1.5, dan IHT3.24) tidak memiliki aktivitas degradasi saponin konsentrasi 0,5 %. Ketiga isolat bakteri tersebut dianalisis kurva pertumbuhannya. Isolat IHT3.24 memiliki laju pertumbuhan paling cepat sehingga digunakan pada uji berikutnya. Perlakuan kombinasi isolat IHT3.24 dan saponin mampu menurunkan TPH minyak bumi sebesar 38 % setelah inkubasi 30 hari. Nilai ini lebih tinggi daripada nilai perlakuan yang hanya diberi bakteri yaitu sebesar 20 %. Data ini menunjukkan bahwa kombinasi bakteri dan saponin lebih efektif dalam mendegradasi minyak bumi dibandingkan dengan ix degradasi minyak bumi oleh bakteri tanpa pemberian saponin. Isolat bakteri IHT1.3, IHT1.5, dan IHT3.24 berdasarkan sekuen 16S rDNA secara berurutan teridentifikasi sebagai Ochrobactrum pseudogrignonense (99,6 %), Pseudomonas mendocina (99,2 %), dan Ochrobactrum pituitosum (97,8 %). Hasil penelitian ini menunjukkan bahwa penambahan saponin Sapindus rarak lebih efektif meningkatkan kemampuan bakteri hidrokarbonoklastik dalam mendegradasi minyak bumi dari Desa Wonocolo hingga dua kali lipat dibanding bakteri tanpa pemberian saponin. Saponin sebagaimana surfaktan diketahui mampu mensolubilisasi minyak bumi sehingga bakteri dapat lebih cepat menyerap substrat tersebut. Kombinasi tersebut dapat dikembangkan sebagai agen bioremediasi ekosistem tercemar minyak bumi
English Abstract
Surfactants have an important role in enhancing the biodegradation of petroleum. A natural surfactant that is rarely explored in helping biodegradation of petroleum waste is saponins. The purpose of this research is to understand the effect of combining saponin from Sapindus rarak plant with isolated hidrocarbonoclastic bacterium in decreasing the level of total petroleum hydrocarbons. Hydrocarbonoclastic bacteria were isolated from crude-oil contaminated soil at Wonocolo Village, Bojonegoro Regency, East Java Province. Bacteria were isolated using enrichment culture method with three consecutive stages in Bushnell-Haas broth medium containing 0.5 % crude oil. Isolated bacteria were screened within two stages. The first stage, isolates were inoculated into nutrient broth medium containing 0 %, 8 %, and 12 % saponin to obtain saponin-tolerant isolates. The second stage, isolates were inoculated into Bushnell- Haas broth medium containing 0.5 % saponin to obtain isolates incapable of degrading saponin. The isolates that passed the screening were made their growth curves in Bushnell- Haas broth medium with 0.5 % crude oil and selected according to the fastest growth rate. Selected isolate was used in biodegradation test of 0.5 % crude oil in Bushnell-Haas broth medium. The test was done with factorial Complete Random Design with two treatments, i.e. existence of isolated bacterium and 0.5 % saponin. Reduction of crude-oil total petroleum hydrocarbons (TPH) was calculated based on the level of TPH. Data of TPH were statistically analyzed using Analysis of Variance at the level of α 0.05. Superior bacteria were identified based on 16S rDNA sequences and constructed in the form of phylogeny trees. Thirty-four bacterial isolates were obtained from the crude-oil contaminated soil. Nine isolates (IHT1.3, IHT1.5, IHT3.8, IHT3.14, IHT3.17, IHT3.19, IHT3.20, IHT3.23, and IHT3.24) were highly tolerance to 12 % saponins. Among them, three isolates (IHT1.3, IHT1.5, and IHT3.24) were unable to degrade saponin at a concentration of 0.5 %. Growth curves of those isolates were analyzed. Isolate IHT3.24 showed the fastest growth rate so that it was used on the next test. A treatment that combining isolate IHT3.24 and saponin showed 38 % reduction of crude oil TPH within 30 days. The value was higher than that used the isolate without addition of saponin that was 20 %. This data demonstrated that a combination of a bacterium and saponin was more effective in degrading crude oil than a bacterium it self without saponin. Isolates of IHT1.3, IHT1.5, and IHT3.24 were identified as Ochrobactrum pseudogrignonense (99,6 %), Pseudomonas mendocina (99,2 %), and Ochrobactrum pituitosum (97,8 %), respectively. xi The result of this experiment indicated that addition of Sapindus rarak saponin improved the ability of a hydrocarbonoclastic bacterium in degradation of crude oil twice as much as the bacterium without saponin. Saponins as surfactant act as crude-oil solubilizing agents so that the bacterium can uptake crude oil faster
Other obstract
-
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/628.5/FMIPA/s/2019/041903802 |
| Subjects: | 600 Technology (Applied sciences) > 628 Sanitary engineering > 628.5 Pollution control and industrial sanitation |
| Divisions: | S2/S3 > Magister Biologi, Fakultas MIPA |
| Depositing User: | Nur Cholis |
| Date Deposited: | 22 Aug 2022 02:05 |
| Last Modified: | 22 Aug 2022 02:05 |
| URI: | http://repository.ub.ac.id/id/eprint/193399 |
![[thumbnail of EVI OCTAVIANY.pdf]](http://repository.ub.ac.id/style/images/fileicons/text.png) |
Text
EVI OCTAVIANY.pdf Download (6MB) |
Actions (login required)
 |
View Item |