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Identifikasi Molekuler dan Analisis Filogenetik Khamir Penghasil Inulinase dari Nira Siwalan (Borassus flabellifer) Berdasarkan Sekuens Internal Transcribed Spacer (ITS)
ABSTRAK
Dewi Ayu Maryati, 24020116120040. Identifikasi Molekuler dan Analisis
Filogenetik Khamir Penghasil Inulinase dari Nira Siwalan (Borassus flabellifer)
Berdasarkan Sekuens Internal Transcribed Spacer (ITS). Dibimbing oleh
Wijanarka dan Rejeki Siti Ferniah.
Inulinase (E.C. 3.2.1.7) adalah enzim yang mampu menghidrolisis inulin
menjadi monomer fruktosa dan biasanya dihasilkan oleh mikroorganisme seperti
khamir. Penelitian ini menggunakan isolat khamir N1 dan N4 yang diisolasi dari
nira siwalan (Borassus flabellifer) yang diketahui dapat menghasilkan inulinase.
Tujuan dari penelitian ini yaitu untuk mengidentifikasi secara molekuler khamir
inulinulitik N1 dan N4 serta hubungan kekerabatannya dengan spesies lain
berdasarkan sekuens internal transcribed spacer (ITS). Tahapan dalam penelitian
ini yaitu isolasi DNA khamir menggunakan metode chelex, amplifikasi DNA
dengan primer ITS 4 dan ITS 5, elektroforesis, analisis sekuensing dan
penyusunan pohon filogenetik. Hasil isolasi DNA menunjukkan konsentrasi DNA
khamir N1 495,8 ng/μl dengan kemurnian 2,12 dan konsentrasi DNA khamir N4
90,2 ng/μl dengan kemurnian 2,32. Produk PCR dari amplifikasi fragmen ITS
menghasilkan ukuran pita ± 500 bp untuk khamir N1 dan ± 600 bp untuk khamir
N4. Hasil BLAST menunjukkan bahwa isolat khamir N1 memiliki homologi
99,45% dengan Candida parapsilosis dan khamir N4 memiliki homologi 97,68%
dengan Candida mucifera. Analisis filogenetik menunjukkan bahwa isolat khamir
N1 dan N4 tidak berkerabat dekat (berbeda klade). Khamir N1 berkerabat dekat
dengan MK638869.1 C. parapsilosis dan khamir N4 berkerabat dekat dengan
NR130678.1 C. mucifera.
Kata kunci: Siwalan (Borassus flabellifer), Inulinase, Khamir, ITS
ABSTRACT
Dewi Ayu Maryati, 24020116120040. Molecular Identification and Phylogenetic
Analysis of Inulinase-Producing Yeast from Nira Siwalan (Borassus flabellifer)
Based on Internal Transcribed Spacer (ITS) Sequences. Supervised by Wijanarka
and Rejeki Siti Ferniah.
Inulinase (E.C. 3.2.1.7) is an enzyme that is able to hydrolyze inulin to be a
fructose monomer and is usually produced by microorganisms such as yeast. This
research uses yeast N1 and N4 isolates isolated from siwalan juice (Borassus
flabellifer) which is known to produce inulinase. The purpose of this study is to
identify molecularly the inulinulitic yeast N1 and N4 and their kinship
relationships with other species based on internal transcribed spacer (ITS)
sequences. The method in this research are yeast DNA isolation using the chelex
method, DNA amplification with primers ITS 4 and ITS 5, electrophoresis,
sequencing analysis and phylogenetic trees preparation. The results of DNA
isolation showed the concentration of yeast DNA N1 495.8 ng/ μl with purity of
2.12 and the concentration of yeast DNA N4 of 90.2 ng/ μl with purity of 2.32.
The PCR product from the ITS fragment amplification produced a band size of ±
500 bp for N1 yeast and ± 600 bp for N4 yeast. Results of BLAST showed that
the isolates of Yeast N1 had homology 99.45% with Candida parapsilosis and
Yeast N4 had homology 97.68% with Candida mucifera. Phylogenetic analysis
showed that the isolates of yeast N1 and N4 were not closely related (different
clade). Yeast N1 is closely related to MK638869.1 C. parapsillosis and Yeast N4
is closely related to NR130678.1 C. mucifera.
Keywords: Siwalan (Borassus flabellifer), Inulinase, Yeast, ITS
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