Kusuma, Nava Indah and Nasrullah Bai Arifin,, S.Pi., M.Sc. and Aulia Rahmawati,, S.P., M.Sc. (2023) Identifikasi Isolat Thraustochytrids dari Habitat Mangrove di Kabupaten Sidoarjo Sebagai Penghasil Asam Lemak. Sarjana thesis, Universitas Brawijaya.
Abstract
Produksi lemak dari mikroalga secara komersil dapat menggantikan minyak ikan sebagai produsen utama omega-3. Kebutuhan minyak ikan di Indonesia sebesar 338.932 kg diikuti permintaan pasar minyak ikan mencapai 10.056.744 kg. Thraustochytrids merupakan salah satu mikroorganisme dengan kandungan lipid tinggi, dapat ditemukan pada ekosistem mangrove dan laut. Produksi lipid dari Thrausrochytrids ini dapat mencapai 80% dari berat kering sel. Meninjau potensi dari Thraustochytrids perlu adanya uji asam lemak untuk mengetahui senyawa yang terdapat pada speies yang diidentifikasi. Penelitian dilaksanakan pada bulan Februari – Maret 2023 di Laboratorium Parasit dan Penyakit Ikan Universitas Brawijaya Malang dilanjutkan sekuensing DNA di PT. Genetika Science Indonesia. Analisis profil asam lemak dilakukan pengujian di Universitas Widyamandala Surabaya. Metode penelitian yang digunakan yaitu deskriptif dengan meneliti 4 sampel isolat Thraustrochytrids. Kegiatan penelitian meliputi identifikasi secara morfologi dan molekuler dilanjutkan dengan uji asam lemak Thraustochytrids dengan prosedur yang diterapkan pada tempat pengujian. Kultur Thraustochytrids pada media padat YPG. Identifikasi morfologi dilakukan pengamatan jaringan ektoplasma, bentuk dan ukuran sel vegetatif, bentuk dan ukuran zoosporangium serta dinding sel. Identifikasi molekuler dengan menggunakan PCR (Polymerase Chain Reaction) dengan set primer 18S Univ F dan 18S Univ R dilanjutkan sekuensing DNA. Tahap PCR meliputi ekstraksi, amplifikasi dengan primer dan elektroforesis. Isolat yang diuji terdeteksi adanya Thraustochytrids sehingga dilanjutkan sekuensing DNA. Pengujian asam lemak dilakukan untuk mengetahui senyawa dalam isolat yang diuji. Uji asam lemak menggunakan Direct Transesterification dan dianalisis menggunakan Gas Cromatography and Mass Spectroscopy (GC- MS). Hasil penelitian ini menunjukkan keempat sampel uji ditemukan spesies Aurantiochytrium acetophilum dengan panjang pita DNA berkisar antara 1.700 base pair – 2.100 base pair. Nilai presentase kesamaan (% identity) yang didapatkan yaitu pada isolat 1 sebesar 94,15%, isolat 2 sebesar 95,15%, isolat 3 sebesar 95,22%, isolat 4 sebesar 95,45%. Hasil uji profil asam lemak masing- masing isolat 1, 2,3 menghasilkan 2 senyawa asam lemak dan pada isolat 4 menghasilkan 20 asam lemak. Hasil analisis didominasi oleh asam lemak esensial yang merupakan inti dari kandungan Thraustrochytrids. Berdasarkan penelitian ini dapat disimpulkan bahwa identifikasi 4 isolat berdasarkan morfologi dan molekuler ditemukan spesies Aurantiochytrium acetophilum. Isolat kemudian diuji asam lemaknya dengan metode direct transesterifikasi menggunakan GC-MS. Terdeteksi adanya asam lemak esensial yaitu DHA terdapat pada isolat 4.Perbedaan hasil asam lemak dapat terjadi dikarenakan kandungan yang terdapat pada sampel uji terlalu sedikit sehingga tidak dapat terdeteksi.
English Abstract
Commercial production of fats from microalgae can replace fish oil as the main producer of omega-3. The need for fish oil in Indonesia is 338,932 kg, followed by market demand for fish oil reaching 10,056,744 kg. Thraustochytrids are one of the microorganisms with high lipid content, which can be found in mangrove and marine ecosystems. Lipid production from Thrausrochytrids can reach 80% of the dry weight of the cell. Reviewing the potential of Thraustochytrids, it is necessary to carry out a fatty acid test to determine the compounds present in the identified species. The research was conducted in February - March 2023 at the Laboratory of Parasites and Fish Diseases, University of Brawijaya Malang, followed by DNA sequencing at PT. Indonesian Science Genetics. Analysis of the fatty acid profile was carried out at Widyamandala University, Surabaya. The research method used was descriptive by examining 4 samples of Thraustrochytrids isolates. Research activities included morphological and molecular identification followed by testing for Thraustochytrids fatty acids with procedures applied at the test site. Thraustochytrids culture on YPG solid media. Morphological identification was carried out by observing the ectoplasmic tissue, the shape and size of the vegetative cells, the shape and size of the zoosporangium and the cell wall. Molecular identification using PCR (Polymerase Chain Reaction) with primer sets 18S Univ F and 18S Univ R followed by DNA sequencing. The PCR stage includes extraction, amplification with primers and electrophoresis. The tested isolates detected the presence of Thraustochytrids so that DNA sequencing was continued. Fatty acid testing was carried out to determine the compounds in the isolates tested. Fatty acid test using Direct Transesterification and analyzed using Gas Chromatography and Mass Spectroscopy (GC-MS). The results of this study indicated that the four test samples were found to be Aurantiochytrium acetophilum species with DNA band lengths ranging from 1,700 base pairs – 2,100 base pairs. The percentage of similarity (% identity) obtained was isolate 1 of 94.15%, isolate 2 was 95.15%, isolate 3 was 95.22%, isolate 4 was 95.45%. The results of the fatty acid profile test for each isolate 1, 2,3 produced 2 fatty acid compounds and isolate 4 produced 20 fatty acids. The results of the analysis are dominated by essential fatty acids which are the essence of the Thraustrochytrids. Based on this study it can be concluded that the identification of 4 isolates based on morphology and molecular found Aurantiochytrium acetophilum species. The isolates were then tested for their fatty acids using the direct transesterification method using GC-MS. The presence of essential fatty acids, namely DHA, was detected in isolate 4. Differences in the yield of fatty acids could occur because the content contained in the test sample was too small so that it could not be detected.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | 0523080179 |
| Subjects: | 600 Technology (Applied sciences) > 639 Hunting, fishing & conservation > 639.3 Culture of cold-blooded vertebrates |
| Divisions: | Fakultas Perikanan dan Ilmu Kelautan > Budidaya Perairan |
| Depositing User: | soegeng sugeng |
| Date Deposited: | 14 Sep 2023 01:49 |
| Last Modified: | 14 Sep 2023 01:49 |
| URI: | http://repository.ub.ac.id/id/eprint/202921 |
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