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Title: | Ekspresi Gen INO1 pada Khamir Wickerhamomyces anomalus yang Terpapar Cekaman Etanol |
Other Titles: | The Expression of INO1 Gene in Wickerhamomyces anomalus Yeast Exposed to Ethanol Stress |
Authors: | Astuti, Rika Indri Putra, Ivan Permana Pradipthawan, Ahmad Zahrani |
Issue Date: | 2024 |
Publisher: | IPB University |
Abstract: | Bioetanol merupakan salah satu sumber energi terbarukan yang dapat diproduksi oleh khamir melalui proses fermentasi. Namun, proses ini sering menghadapi tantangan berupa cekaman etanol yang dapat menghambat pertumbuhan sel khamir. Penelitian ini menganalisis tingkat ekspresi gen INO1 pada Wickerhamomyces anomalus isolat BT2 dan BT5 dalam kondisi cekaman etanol untuk memahami mekanisme toleransi etanol pada khamir tersebut. Gen INO1 menyandikan myo-inositol-1-phosphate synthase yang terlibat dalam toleransi khamir terhadap cekaman etanol. Validasi toleransi etanol dilakukan secara kualitatif menggunakan metode spot test, diikuti dengan analisis reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) untuk mengukur tingkat ekspresi gen INO1. Hasil spot test menunjukkan bahwa BT2 memiliki toleransi tertinggi terhadap etanol hingga konsentrasi 10%, diikuti oleh BT5 dan S. cerevisiae sebagai pembanding. Analisis RT-qPCR mengungkapkan bahwa ekspresi gen INO1 pada BT2 meningkat hampir 14,3 kali lipat dalam kondisi cekaman etanol 10%, sementara BT5 dan S. cerevisiae mengalami penurunan ekspresi gen INO1. Hal ini menunjukkan bahwa toleransi etanol pada BT2 diduga diregulasi oleh gen INO1, sedangkan pada BT5 dan S. cerevisiae kemungkinan diregulasi oleh gen lain yang mempengaruhi kemampuannya untuk toleran terhadap cekaman etanol. Bioethanol is a renewable energy source that can be produced by yeast through the fermentation process. However, this process often faces challenges such as ethanol stress which can inhibit yeast cell growth. This study analyzes the level of INO1 gene expression in Wickerhamomyces anomalus isolates BT2 and BT5 under ethanol stress conditions to understand the ethanol tolerance mechanism in these yeasts. The INO1 gene encodes myo-inositol-1-phosphate synthase, which is involved in yeast tolerance to ethanol stress. Ethanol tolerance validation was performed qualitatively using the spot test method, followed by reverse transcriptase- quantitative polymerase chain reaction (RT-qPCR) analysis to measure the level of INO1 gene expression. Spot test results showed that BT2 had the highest tolerance to ethanol up to the concentration of 10%, followed by BT5 and S. cerevisiae as a comparison. RT-qPCR analysis revealed that INO1 gene expression in BT2 increased nearly 14,3-fold under 10% ethanol, while BT5 and S. cerevisiae experienced a decrease in INO1 gene expression. These results indicate that the INO1 gene likely regulates ethanol tolerance in BT2, whereas in BT5 and S. cerevisiae, other genes are possibly responsible for regulating their ability to tolerate ethanol stress. |
URI: | http://repository.ipb.ac.id/handle/123456789/155653 |
Appears in Collections: | UT - Biology |
Files in This Item:
File | Description | Size | Format | |
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cover_G3401201066_e0fbf0ca25864d61a2c7ea4dd28255ee.pdf | Cover | 298.42 kB | Adobe PDF | View/Open |
fulltext_G3401201066_00caa986dbb94d2db50487dc939af2a0.pdf Restricted Access | Fulltext | 1.26 MB | Adobe PDF | View/Open |
lampiran_G3401201066_f0af028cdca54733b597278986b7039a.pdf Restricted Access | Lampiran | 154.92 kB | Adobe PDF | View/Open |
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