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Identifikasi Molekuler Isolat Monascus sp. Hasil Isolasi Angkak Berdasarkan Gen Internal Transcribed Spacer (ITS) dan Pengukuran Kandungan Pigmen
ABSTRAK
Identifikasi kapang Monascus berdasarkan karakter morfologi rentan terhadap kesalahan
identifikasi, karena persamaan bentuk dan warna. Dewasa ini, identifikasi molekuler telah
dilakukan untuk melengkapi identifikasi secara morfologi, misalnya identifikasi kapang
menggunakan gen Internal Transcribed Spacer (ITS). Tujuan dari penelitian ini yaitu
mengidentifikasi isolat kapang Monascus yang diisolasi dari angkak dengan menggunakan
gen Internal Transcribed Spacer (ITS), melakukan analisis hubungan kekerabatan,
melakukan pengukuran pertumbuhan sel dan produksi pigmen. Sekuens ITS Monascus
diamplifikasi menggunakan primer ITS 1 dan ITS 4, sedangkan analisis filogenetik molekuler
menggunakan metode Neighbor Joining (NJ). Berdasarkan persamaan homologi melalui
metode Basic Local Alignment Search Tool (BLAST) dan analisis pohon filogenetik, isolat
kapang teridentifikasi sebagai Monascus purpureus sebesar 100%. Pengukuran pertumbuhan
sel kapang Monascus dilakukan menggunakan berat kering sel. Pengukuran pigmen
dilakukan menggunakan spectrophotometer dengan panjang gelombang 390 nm untuk
pigmen kuning dan 500 nm untuk pigmen merah. Pengukuran produksi pigmen mengalami
peningkatan hingga hari ke-13. Jumlah produksi pigmen ekstraseluler warna kuning
sebesar 37,358 U/g; sedangkan pigmen warna merah sebesar 2,6545 U/g. Jumlah
produksi pigmen intraseluler warna merah sebesar 7,4175 U/g, sedangkan pigmen
warna kuning 30,176 U/g.
Kata kunci: Monascus, angkak, ITS, filogenetik molekuler, pigmen
ABSTRACT
Identification of Monascus based on their morphological characteristic, is problematic due to
several specimens shared similarity on morphology and colours. Nowadays, molecular
identification has been done to complement morphological identification, for example mold
identification using the Internal Transcribed Spacer (ITS) gene. The aims of study were to
identify Monascus isolates resulted from angkak using Internal Transcribed Spacer (ITS)
gene, phylogenetic analyzing, measurement cell growth and production of pigment.
Sequences of ITS Monascus was amplified using ITS 1 and ITS 4 primer, whereas the
molecular phylogenetic analyses was done using Neighbor Joining (NJ) method. Based on
homology search by Basic Local Alignment Search Tool (BLAST) program and phylogenetic
tree analyses, Monascus isolate was identified as Monascus purpureus (100%). Measurement
of Monascus cell growth was carried out measuring dry weight cell. Measurement of pigment
was done using spectrophotometer at 390 nm wavelength for yellow pigment and 500 nm for
red pigment. The measurement of pigments production showed increased content until day
13. The amount of yellow extracellular pigment production was 37.358 U / g; whereas the red
color pigment was 2.6545 U / g. The amount of red intracellular pigment production was
7.4175 U / g, while the yellow pigment was 30.176 U / g.
Keywords: Monascus, angkak, molecular phylogenetic, pigment
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