Pretreatment Selulase Biomassa Mikroalga (Nannochloropsis oculata) untuk Meningkatkan Efektivitas Ekstraksi Protein dengan Metode Ultrasonikasi

Kharisma, Arnelita (2026) Pretreatment Selulase Biomassa Mikroalga (Nannochloropsis oculata) untuk Meningkatkan Efektivitas Ekstraksi Protein dengan Metode Ultrasonikasi. Other thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Nannochloropsis oculata merupakan mikroalga dengan kandungan protein tinggi yang berpotensi sebagai sumber protein alternatif. Namun, dinding selnya yang didominasi selulosa hingga 80% dari total polisakarida menjadi hambatan utama dalam proses ekstraksi protein. Selulase dari Trichoderma harzianum merupakan enzim yang mampu mendegradasi selulosa secara spesifik tanpa menyebabkan denaturasi protein, sehingga berpotensi digunakan sebagai agen pretreatment untuk meningkatkan efisiensi ekstraksi protein mikroalga, namun pengaruh variasi volume selulase terhadap degradasi dinding sel dan kadar protein hasil ekstraksi pada N. oculata melalui metode ultrasonikasi belum pernah dieksplorasi. Penelitian ini bertujuan untuk mengetahui pengaruh variasi volume selulase T. harzianum pada tahap pretreatment terhadap kadar gula pereduksi dan kadar protein hasil ekstraksi biomassa N. oculata melalui metode ultrasonikasi. Selulase diproduksi melalui submerged fermentation dengan media yang mengandung carboxymethylcellulose (CMC) sebagai sumber karbon melalui tahapan kultur starter, aklimatisasi, dan produksi. Sebagian crude selulase dikarakterisasi melalui pemurnian parsial menggunakan amonium sulfat serta penentuan titik isoelektrik, sedangkan sisanya diaplikasikan untuk pretreatment biomassa kering N. oculata dengan variasi volume enzim 0, 50, 100, dan 150 µL. Biomassa hasil pretreatment selanjutnya diekstraksi menggunakan metode ultrasonikasi dengan ultrasonic cell disrupter BIOBASE UCD-500. Parameter yang diamati meliputi kadar gula pereduksi dan kadar protein hasil ekstraksi. Data dianalisis menggunakan ANOVA satu arah dengan uji lanjut Tukey (p<0,05). Hasil penelitian menunjukkan bahwa variasi volume selulase berpengaruh signifikan terhadap kedua parameter yang diamati. Kadar gula pereduksi meningkat dari 92,67±3,33 mg/mL pada kontrol menjadi 1956,0±24,2 mg/mL pada volume enzim 150 µL, mengindikasikan terjadinya hidrolisis selulosa dinding sel secara progresif. Kadar protein hasil ekstraksi juga meningkat dari 2,65±0,17 mg/mL menjadi 11,49±0,50 mg/mL, setara dengan peningkatan 4,33 kali lipat. Volume enzim 150 µL merupakan perlakuan optimal yang menghasilkan kadar protein dan kadar gula pereduksi tertinggi, menunjukkan bahwa kombinasi pretreatment selulase dan ekstraksi ultrasonikasi merupakan strategi yang efektif untuk meningkatkan efisiensi ekstraksi protein dari biomassa N. oculata.
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Nannochloropsis oculata is a microalga with high protein content that holds potential as an alternative protein source. However, the high cellulose content of its cell wall, accounting for up to 80% of total polysaccharides, poses a major barrier to efficient protein extraction. Cellulase from Trichoderma harzianum is an enzyme capable of specifically degrading cellulose without causing protein denaturation, making it a promising agent for pretreatment to enhance the efficiency of microalgal protein extraction; however, the effect of cellulase volume variation on cell wall degradation and extracted protein content in N. oculata via ultrasonication has never been explored. This study aimed to investigate the effect of varying T. harzianum cellulase volumes during the pretreatment stage on reducing sugar content and extracted protein content from N. oculata biomass using the ultrasonication method. Cellulase was produced via submerged fermentation using a medium containing carboxymethylcellulose (CMC) as the carbon source, through the stages of starter culture, acclimatization, and production. A portion of the crude cellulase was characterized through partial purification using ammonium sulfate precipitation and isoelectric point determination, while the remainder was applied for pretreatment of dry N. oculata biomass at enzyme volume variations of 0, 50, 100, and 150 µL. The pretreated biomass was subsequently extracted using the ultrasonication method with a BIOBASE UCD-500 ultrasonic cell disrupter. Parameters observed included reducing sugar content and extracted protein content. Data were analyzed using one-way ANOVA followed by Tukey's post hoc test (p<0.05). The results showed that cellulase volume variation had a significant effect on both parameters. Reducing sugar content increased from 92.67±3.33 mg/mL in the control to 1956.0±24.2 mg/mL at an enzyme volume of 150 µL, indicating progressive hydrolysis of cell wall cellulose. Extracted protein content also increased from 2.65±0.17 mg/mL to 11.49±0.50 mg/mL, representing a 4.33-fold increase. An enzyme volume of 150 µL was identified as the optimal treatment, yielding the highest protein and reducing sugar contents, demonstrating that the combination of cellulase pretreatment and ultrasonication extraction is an effective strategy for improving protein extraction efficiency from N. oculata biomass.

Item Type:	Thesis (Other)
Uncontrolled Keywords:	Nannochloropsis oculata, Selulase, Submerged fermentation, Trichoderma harzianum, Ultrasonikasi, Nannochloropsis oculata, Cellulase, Submerged fermentation, Trichoderma harzianum, Ultrasonication.
Subjects:	Q Science > QH Biology > QH301 Biology Q Science > QK Botany > QK568 Microalgae Q Science > QR Microbiology
Divisions:	Faculty of Science and Data Analytics (SCIENTICS) > Biology > 46201-(S1) Undergraduate Thesis
Depositing User:	Arnelita Kharisma
Date Deposited:	06 Jul 2026 00:45
Last Modified:	06 Jul 2026 00:45
URI:	http://repository.its.ac.id/id/eprint/134297

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