Isolasi dan Analisis In-Silico Fragmen cDNA Fatty Acid Desaturase 2 (Fad2) Kemiri Sunan (Reutealis Trisperma)

Prasetyowati, Indah (2018) Isolasi dan Analisis In-Silico Fragmen cDNA Fatty Acid Desaturase 2 (Fad2) Kemiri Sunan (Reutealis Trisperma). Undergraduate thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Komposisi asam lemak dominan pada minyak Kemiri Sunan (R. trisperma) adalah asam stearat (9%), asam palmitat (10%), asam oleat (12%), asam linoleat (19%), serta asam α-eleostearat (51%). Kandungan asam lemak tidak jenuh jamak (PUFA), yaitu asam linoleat dan asam α-eleostearat, dapat menurunkan stabilitas oksidasi biodiesel R. trisperma. Beberapa studi menunjukkan bahwa enzim Fatty Acid Desaturase 2 (FAD2) berperan dalam regulasi desaturasi asam lemak, namun informasi genetik FAD2 R. trisperma hingga saat ini masih belum diketahui. Penelitian ini bertujuan untuk mengisolasi, mengkarakterisasi, serta mengetahui hubungan fragmen FAD2 R. trisperma dengan spesies Euphorbiaceae yang lain. Isolasi fragmen cDNA dilakukan dengan pendekatan Reverse Transkriptase-PCR. Sekuen DNA diperoleh melalui sekuensing kemudian dianalisis secara in-silico untuk mengetahui identitas fragmen, lokasi subseluler, serta hubungan kekerabatan cDNA FAD2 R. trisperma. Hasil penelitian menunjukkan bahwa sekuen parsial FAD2 R. trisperma yang berhasil diisolasi adalah sebesar 923 bp. Karakteristik FAD2 setelah dianalisis secara in-silico diprediksi mengkode 260 asam amino, memiliki similaritas domain dengan Omega-6 fatty acid desaturase, serta berlokasi pada membran retikulum endoplasma. Fragmen gen FAD2 R. trisperma menunjukkan kedekatan dengan Vernicia fordii (HM755946.1). ============= The predominant fatty acid of Kemiri Sunan (R. trisperma) are stearic acid (9%), palmitic acid (10%), oleic acid (12%), linoleic acid (19%), and α-eleostearic acid (51%). The presence of polyunsaturated fatty acid (PUFA), linoleic acid and α-eleostearic acid, will decrease the oxidation stability of R. trisperma biodiesel. Although several studies have suggested that fatty Acid Desaturase 2 (FAD2) enzyme is involved in the regulation of fatty acid desaturation, little is known about the genetic information of FAD2 in R. trisperma. The objectives of this study were to isolate, characterize, and determine the relation of R. trisperma FAD2 fragment among Euphorbiaceae. Isolation of cDNA fragment was performed by Reverse Transcriptase-PCR. The DNA sequence obtained through sequencing used for further analysis. In-silico analysis included identify the fragment identity, subcellular localization, phylogenetic construction between R. trisperma FAD2 cDNA fragment and Euphorbiaceae. The results showed that 923 bp partial sequence of R. trisperma FAD2 was successfully isolated. Based on in-silico analysis, the characteristics of FAD2 were predicted encode 260 amino acids, had a domain similarity with Omega-6 fatty acid desaturase, and located in the endoplasmic reticulum membrane. R. trisperma FAD2 fragment was more closely related to Vernicia fordii (HM755946.1).

Item Type: Thesis (Undergraduate)
Uncontrolled Keywords: biodiesel; Fatty Acid Desaturase; in-silico; Reutealis trisperma
Subjects: Q Science > QK Botany
Q Science > QP Physiology > QP624 Molecular biology.
Divisions: Faculty of Mathematics and Science > Biology > 46201-(S1) Undergraduate Thesis
Depositing User: Indah Prasetyowati
Date Deposited: 26 Jan 2018 02:19
Last Modified: 26 Jan 2018 02:19
URI: http://repository.its.ac.id/id/eprint/50729

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