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All Journal Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences Mutiara Medika: Jurnal Kedokteran dan Kesehatan Emerging Science Journal
BUDI SAKSONO
Research Center for Biotechnology, Lembaga Ilmu Pengetahuan Indonesia
Agriculture, Biological Sciences & Forestry Environmental Science Health Professions Medicine & Pharmacology

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The Cytotoxicity of Agaro-Oligosaccharides and Neoagaro-Oligosaccharides on Macrophage Cells Pratitis, Visi Endah; Puspitasari, Pinki Anggrahini; Hanbal, Mayland Muhammad; Tsabitah, Khansa; Juliadmi, Dian; Saksono, Budi; Wijayanti, Nastiti
Mutiara Medika: Jurnal Kedokteran dan Kesehatan Vol 24, No 2 (2024): July
Publisher : Universitas Muhammadiyah Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18196/mmjkk.v24i2.21059

Abstract

Agarose is a polysaccharide from red algae. Enzymatic hydrolysis of agarose can produce Agaro-Oligosaccharidess (AOS) and Neoagaro-Oligosaccharidess (NAOS). Different times of enzymatic hydrolysis can produce different types of AOS and NAOS based on their degree of polymerization (DPs). This study aims to examine the cytotoxicity of AOS and NAOS with different hydrolysis times on the macrophage cell line RAW 264.7. The parameters used were the percentage of cell viability and IC50 value. The cytotoxicity test using MTT assay and One-way ANOVA were used as statistical tests. The results showed that AOS-0 at a concentration below 125 µg/mL was not toxic and showed moderate toxicity up to a high concentration of 1000 µg/mL, while AOS hydrolyzed for 24 h (AOS-24) was not toxic to RAW 264.7 cells at all concentrations tested. The different results were shown in all NAOS samples, which were highly toxic to RAW 264.7 cells in the 125 to 1000 µg/mL, indicating that it was concentration dependent. The results showed that different hydrolysis times caused differences in the structure of AOS and NAOS compounds and influenced the toxicity level. Research development for further studies on antioxidants and anti-inflammatory needs more attention to the sample type and hydrolysis time.
In-Silico Optimization of α-agarase for Sustainable Bioprospecting of Agaro-oligosaccharides: In-Silico Optimization of α-agarase for Sustainable Bioprospecting of Agaro-oligosaccharides Hanbal, Mayland Muhammad; Wijayanti, Nastiti; Tsabitah, Khansa; Zulfa, Amania; Tasania, Nadia Nisa; Saksono, Budi
Journal of Tropical Life Science Vol. 15 No. 2
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/gg6mkk56

Abstract

Enzymatic synthesis is a sustainable alternative to chemical methods, offering high substrate specificity, reduced byproducts, and environmentally friendly processes. Despite its advantages, agaro-oligosaccharides (AOS) production largely depends on chemical synthesis due to the limited exploration of α-agarase. Therefore, this study aims to identify, analyze, and redesign a suitable α-agarase candidate for AOS production using in-silico approaches. Biological databases from CAZy, UniProtKB, and PDB, along with journal references, were used to curate α-agarase candidates. Non-catalytic regions were removed to retain only the GH96 catalytic domain, with a His-tag added for easier purification. Subsequently, structural modeling using SWISS-MODEL was performed to facilitate blind docking with CB-Dock2. Modeling also facilitated physicochemical properties predictions incorporating OphPred, Protein-Sol, and SCooP for pH, solubility, and thermal stability. The results showed that AgaA33, obtained from Thalassomonas agarivorans JAMB-A33, was selected due to its high annotation score and optimal temperature. Structural modeling and blind docking confirmed that the functional domains were preserved after redesign. In-silico physicochemical assessments revealed that the redesigned enzyme exhibited improved pH tolerance and thermal stability, despite a slight reduction in solubility. This study showed the use of computational tools for enzyme redesign and showed the potential of α-agarase as a green and sustainable biocatalyst for AOS production. By combining database-driven candidate selection with in-silico structural and functional analyses, these results set the foundation for further optimization of α-agarase to meet industrial needs. Future efforts must focus on improving solubility and refining activity predictions to fully realize the enzyme’s potential for eco-friendly bioprocesses.
Agarose-Based Antibacterial Films from Gracilaria sp.: Isolation, Characterization, and Metal Nanoparticle Incorporation Ahmad, Ahyar; Zainuddin, Rahmaniah; Saksono, Budi; Anita, Sita Heris; Zulfiana, Deni; Ermawar, Riksfardini A.; Arfah, Rugaiyah; Natsir, Hasnah; Karim, Harningsih; Irmawati; Ramli, Siti R.; Karim, Abdul
Emerging Science Journal Vol. 9 No. 5 (2025): October
Publisher : Ital Publication

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.28991/ESJ-2025-09-05-03

Abstract

The incorporated metal nanoparticles in a polysaccharide-based film exhibit efficient antibacterial activity against harmful germs. However, previous studies have used a commercial polysaccharide for their film production. Therefore, this study aimed to develop a natural polysaccharide-based film extracted from the local algae Gracilaria sp. originating from Sinjai Regency, South Sulawesi, Indonesia. Firstly, the polysaccharide agarose was isolated and its properties compared with those of commercial agarose. A present low-cost isolation process produces agarose with 1.8% (w/w) of yield. Results also showed physicochemical properties similar to those of the commercial agarose. Secondly, the agarose-based antibacterial film was synthesized at 0, 0.5, and 1% glycerol concentrations. The synthesized film was incorporated with silver (Ag) and copper (Cu) nanoparticles (NPs). Morphological, mechanical, and physicochemical properties of the incorporated Ag-agarose and Cu-agarose films were characterized using Field Emission Scanning Electron Microscope (FESEM), Universal Testing Machine (UTM), and Fourier Transform Infrared Spectroscopy (FTIR), respectively. Results showed the film stiffness and tensile strength increased by incorporating either AgNPs or CuNPS. The interaction of AgNPs-agarose most likely involves physical bonds, while the interaction of CuNPs-agarose forms coordination bonds. An antibacterial test showed that the Ag-agarose nanocomposite inhibited the growth of Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus subtilis. In the meantime, Cu-agarose prevented the growth of Staphylococcus aureus. Overall, antibacterial activity was influenced by the interaction between metal nanoparticles and agarose, the concentration of metal nanoparticles, and the film's solubility. An agarose-based antibacterial film from Gracilaria sp. has the potential for use in various applications, including food packaging, pharmaceuticals, and other industries.
Co-Authors A.A. Ketut Agung Cahyawan W Abdul Karim Ahyar Ahmad Arfah, Rugaiyah Deni Zulfiana Ermawar, Riksfardini A. Hanbal, Mayland Muhammad Harningsih Karim Hasnah Natsir Irmawati Juliadmi, Dian Nastiti Wijayanti Pratitis, Visi Endah Puspitasari, Pinki Anggrahini Ramli, Siti R. Sita Heris Anita Tasania, Nadia Nisa Tsabitah, Khansa Wulandari, Puspita Suci Zainuddin, Rahmaniah Zulfa, Amania