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All Journal Jurnal Akuakultur Indonesia Jurnal Ilmu-Ilmu Perairan dan Perikanan Indonesia Journal of Marine and Aquatic Sciences JURNAL PENGABDIAN KEPADA MASYARAKAT Jurnal Perikanan Universitas Gadjah Mada Jurnal Akuakultur Sungai dan Danau Jurnal Riset Akuakultur Jurnal Medik Veteriner Jurnal Penamas Adi Buana Journal of Aquaculture Science Journal of Aquaculture and Fish Health Grouper: Jurnal Ilmiah Perikanan Journal of Marine and Coastal Science Jurnal Litbang Provinsi Jawa Tengah Berkala Penelitian Hayati Journal of Science and Science Education (JoSSEd) Sewagati Depik Jurnal Ilmu-Ilmu Perairan, Pesisir dan Perikanan Depik Jurnal Ilmu-Ilmu Perairan, Pesisir, dan Perikanan Journal of Fisheries & Marine Jurnal Perikanan
Akhmad Taufiq Mukti
Departemen Akuakultur, Fakultas Perikanan Dan Kelautan, Universitas Airlangga, Jl. Mulyorejo, Surabaya
Agriculture, Biological Sciences & Forestry Arts Humanities Astronomy Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Earth & Planetary Sciences Economics, Econometrics & Finance Education Electrical & Electronics Engineering Energy Engineering Environmental Science Immunology & microbiology Languange, Linguistic, Communication & Media Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Physics Public Health Social Sciences Veterinary Other

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PENINGKATAN AKTIVITAS DAN EKSPRESI PROTEIN TOTAL ENZIM SELULASE REKOMBINAN SERTA POTENSI APLIKASINYA PADA PAKAN IKAN Agniny, Siswi Fililmi; Mahasri, Gunanti; Mukti, Akhmad Taufiq; Amin, Muhamad
Jurnal Riset Akuakultur Vol 20, No 3 (2025): September (2025)
Publisher : Politeknik Kelautan dan Perikanan Jembrana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/jra.20.3.2025.257-267

Abstract

Meskipun enzim selulase rekombinan banyak dikembangkan untuk meningkatkan efisiensi pemecahan serat, informasi mengenai bagaimana modifikasi seperti mutasi dan penambahan tag SKIK (Ser-Lys-Ile-Lys) dapat memengaruhi aktivitas enzim dan produksi protein total masih terbatas. Oleh karena itu, penelitian ini bertujuan mengevaluasi pengaruh mutasi dan penambahan tag SKIK terhadap aktivitas dan protein total enzim selulase rekombinan yang diproduksi oleh Escherichia coli BL21 (DE3). Penelitian dilakukan secara eksperimental menggunakan rancangan acak lengkap (RAL) dengan empat perlakuan, yaitu P1= wild type (WT), P2 = WT + SKIK, P3 = mutan, dan P4 = mutan + SKIK, masing-masing dengan empat ulangan. Produksi enzim dilakukan melalui kultur E. coli rekombinan yang membawa plasmid pET-22b-CellE dengan induksi laktosa monohidrat, kemudian dianalisis menggunakan uji aktivitas selulase metode 3,5-dinitrosalisilat (DNS) dan pengukuran protein total dengan metode Bradford. Data dianalisis menggunakan ANOVA satu arah dan uji lanjut Duncan pada tingkat signifikansi 5% (p < 0,05). Hasil penelitian menunjukkan bahwa perlakuan mutan + SKIK menghasilkan nilai tertinggi baik pada aktivitas enzim (10,66 U mL-1) maupun protein total (0,85 mg mL-1). Penambahan SKIK meningkatkan ekspresi dan kelarutan protein, sedangkan mutasi memperbaiki efisiensi katalitik enzim. Kombinasi keduanya memberikan efek sinergis dalam meningkatkan perfoma enzim selulase rekombinan. Penelitian ini merupakan yang pertama mengombinasikan penambahan tag SKIK dengan mutagenesis untuk meningkatkan ekspresi dan aktivitas katalitik enzim selulase pada E. coli. Temuan ini tidak hanya relevan untuk peningkatan produksi enzim pada industri bioteknologi, tetapi juga berpotensi diaplikasikan dalam formulasi pakan ikan. Although recombinant cellulase enzymes have been widely developed to enhance fiber-degradation efficiency, information regarding how modifications such as mutagenesis and the addition of the SKIK (Ser-Lys-Ile-Lys) tag influence enzymatic activity and total protein production remains limited. Therefore, this study aimed to evaluate the effects of mutagenesis and SKIK tagging on the activity and total protein yield of recombinant cellulase expressed in Escherichia coli BL21 (DE3). The research was conducted experimentally using a completely randomized design (CRD) with four treatments: P1 = wild type (WT), P2 = WT + SKIK, P3 = mutant, and P4 = mutant + SKIK, each consisting of four replications. Enzyme production was carried out using recombinant E. coli harboring the pET-22b-CellE plasmid induced with lactose monohydrate, and the resulting enzyme was analyzed for cellulase activity using the 3,5-dinitrosalicylic acid (DNS) method and total protein concentration using the Bradford assay. Data were analyzed using one-way ANOVA followed by Duncan’s post-hoc test at a 5% significance level (p < 0.05). The results showed that the mutant + SKIK treatment produced the highest values for both enzymatic activity (10.66 U mL-1) and total protein (0.85 mg mL-1). The addition of the SKIK tag enhanced protein expression and solubility, whereas mutagenesis improved catalytic efficiency. The combination of both modifications produced a synergistic effect, resulting in superior performance of the recombinant cellulase enzyme. This study is the first to combine SKIK tagging with mutagenesis to enhance the expression and catalytic activity of cellulase in E. coli. These findings are not only relevant for improving enzyme production in biotechnology industries but also hold potential applications in fish feed formulation.
Fish Biodiversity and Substrate Composition on Coral Reef Ecosystem at Penimbangan, Buleleng Regency, Bali Sapto Andriyono; Diah Ayu Safitri; Akhmad Taufiq Mukti; Mike van Keulen
Depik Vol 14, No 2 (2025): JUNE 2025
Publisher : Faculty of Marine and Fisheries, Universitas Syiah Kuala

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Abstract

The Penimbangan is one of the potential locations for underwater tourism in the Regency Buleleng, by unique dive spot. Planning and sustainable coastal development are essential for the continuity ecosystem, sea, and economy. Fish and coral studies in Penimbangan, District Buleleng, occurred from December 2022 to February 2023. Location observation shared four sites of data retrieval. The determination sites study uses purposive sampling based on the spread of coral naturally. Data was collected by Underwater Photo Transect (UPT) for coral and Underwater Visual Census (UVC) for fish. Transect area coral 50m long with quadratic transect (58cmx44cm), and classified categories using CPCe software. Transect area fish of 350 m2 (length 70 m, limit right and left 2.5 m), identified fish to level species. The most studied habitat associations in coral reefs are coral (HC) and fish in coral assemblages. Percentage composition substrate basis in Penimbangan, namely dead coral with algae (106.74), coral (90.27), and sand (70). Lifeform coral is found in as many as ten types; the most are CM (39.00), CE (23.20), and ACS (10.07). A total of 17 families with 70 reef fish species were recorded in this study. The family with the most species is Pomacentridae (25), followed by Chaetodontidae (9) and Labridae (8). The composition of fish will vary according to the type of habitat. Different coral habitats covered by benthic influence the abundance of certain trophic levels of reef fish in an area of water.
Co-Authors A. Shofy Mubarak Adriansyah, Ivan Agniny, Siswi Fililmi Agung Pamuji Rahayu Alim Isnansetyo Alvira Febrianti Pratiwi Anisa, Hosia Anjasmara, L. Dede Arta Satya Arafik Lamadi Ardiyan, Buyung Raga Aulia Ikhsan Syamsuri Aziz, Ahmad Saiful Boedi Setya Rahardja Browijoyo Santanamurti Catur Pujiono Danita Garneda Rezi DARMAWAN SETIA BUDI Denta Batara Kusuma Hendra Destya Twinandia Dewi, Nina Nurmalia Diah Ayu Safitri Dian Pebianti Dyah Hariani Dyah Hariani Edi Rukyanto Endah Sih Prihatini Endang Tri Wahyurini Fachriza Tri Aprilia Falatehan, Naufal Fasya, Arif Habib Firdaus, Fitrul Azizi Fuquh Rahmat Shaleh Gunanti Mahasri gustini, Maria A Herliana Arsianingtyas Imlani, Ainulyakin Isroni, Wahyu Jati, Ilham Shofananda Pramono Kenconojati, Hapsari Kismiyati , Lailatul Lutfiyah, Lailatul Laksmi Sulmartiwi Lia Oktavia Ika Putri Lutfianasari, Lutfianasari Luthfiana Aprilianita Sari M Wahyu Alfian Mahasari, Gunanti Maria A gustini Mariah, Sharven Rao Mike van Keulen Mohammad Faizal Ulkhaq Muhamad Ali Muhamad Amin Muhamad Amin Muhammad Arief Muhammad Arief Mutiara Rahmawati Nafisyah, Ayu Lana Nanik Siti Aminah Naufal Abiyu Pratama Nizar Afiansyah Loekman Panhar, Fio Surya Pratama Semendo Puguh Yugo Wijanarko Pungky Slamet Wisnu Kusuma Puspitasari, Febriyanti Rahmadhani, Desi Rahmahani, Jola Rahmawan, Dani Bagus Rozi Rozi Rr. Juni Triastuti Rr. Juni Triastuti Sabrina, Annisa Nur Safitri, Diah Ayu Saiful Bakhri Salman Aldo Alfaresi Samara, Syifania Hanifah Santanumurti, Muhammad Browijoyo Sapto Andriyono Sari, Luthfiana Aprilanita Sari, Putri Desi Wulan Septuresty Hartri Eka Slamet Wisnu Kusuma, Pungky South, Josie Sri Harweni Sri Subekti Suciyono, Suciyono Sudarno, Sudarno Van Keulen, Mike Veryl Hasan Vivaldy Phaza Muharta Woro Hastuti Setyantini Yudi Cahyoko