Nutritional profiling and in silico analysis of pharmacological activities from local rice Pulu Mandoti fermented with Pleurotus spp.

Pulu Mandoti, a local red rice (Oryza sativa L.) variety popular among Sulawesi residents, has gained recognition for its perceived health benefits, especially as a preferred dietary option for individuals with diabetes or those seeking to prevent obesity. Given the increasing consumption of mushrooms, particularly Pleurotus species, renowned for their nutritional and medicinal attributes, this study delves into the transformative effects of Pleurotus spp. fermentation on Pulu Mandoti, the indigenous rice variety. Proximate analysis disclosed elevated dry matter (91.99 ± 0.61%), crude protein (8.55 ± 0.15%), and crude fat (1.34 ± 0.05%) in Pleurotus cystidiosus fermentation compared to Pleurotus ostreatus and Pleurotus djamor. Concurrently, antioxidant and antidiabetic activities were notably improved in all Pleurotus fermentations. Pulu Mandoti fermented with P. cystidiosus outperformed other treatments, aligning with molecular docking results pinpointing 11-Eicosenoic acid, methyl ester, and butylated hydroxytoluene as optimal interactors with antioxidant receptors 5O0x and 2CKJ. Butylated hydroxytoluene demonstrated interactions with the antidiabetic receptor 2QV4, along with 9-Octadecenoic acid, methyl ester. These compounds, previously unreported in Pleurotus, displayed promising attributes as antioxidants and antidiabetic agents. Furthermore, the investigation delved into the fatty acid profiles, emphasizing the diverse range of potential bioactive compounds in fermented Pulu Mandoti. The findings of this research present a potential functional food rich in natural antioxidants and antidiabetic compounds, highlighting the yet undiscovered capabilities of Pleurotus spp. fermentation in augmenting the nutritional composition and bioactivity of indigenous rice varieties, specifically Pulu Mandoti.

Endophytic Aspergillii and Penicillii from medicinal plants: a focus on antimicrobial and multidrug resistant pathogens inhibitory activity.

The rise of multidrug resistance among microorganisms, where they develop resistance against formerly efficacious drugs, has led to increased disease prevalence and mortality rates, posing a growing challenge. Globally, antibiotic resistance has made a significant impact, causing millions of fatalities each year. Endophytic fungi have gained considerable attention in research due to their potential to produce a wide variety of secondary metabolites, including natural substances with antimicrobial capabilities. The genera Aspergillus and Penicillium stand out as the most prevalent species of endophytic fungi. Filamentous fungi, such as these are responsible for the production of 45% of known microbial metabolites. This review focuses on exploring the bioactive substances produced by endophytic fungi from these two genera, particularly in conjunction with medicinal plants. Emphasis is placed on their antimicrobial activity and their ability to inhibit multidrug-resistant pathogens. As the need for alternative treatments to combat drug-resistant infections continues to grow, endophytic fungi have the potential to provide a valuable source of bioactive molecules for medical applications.

Profile and in silico analysis of metabolite compounds of the endophytic fungus Alternaria alternata K-10 from Drymoglossum piloselloides as antioxidants and antibacterials.

Endophytic fungi are known for producing secondary metabolites with valuable biological activities, including antiviral, anticancer, antibacterial, and antioxidant properties. This study aims to evaluate an endophytic fungus from Dragon Scales leaves (Drymoglossum piloselloides) and analyze its metabolites as antioxidants and antibacterials. In this study, an endophytic fungus was isolated from the leaves of Dragon Scales (D. piloselloides) and identified using molecular analysis of the Internal Transcribed Spacer (ITS) ribosomal RNA locus. The fungus was authenticated as Alternaria alternata strain K-10. Crude extracts were obtained using n-hexane and ethyl acetate and analyzed via GC-MS Shimadzu-QP 2010 Ultra with NIST spectral library. Antibacterial activity was observed against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa using the paper disc method, showing inhibition zones of 8.7-9.3 mm and 8.8-9.4 mm for ethyl acetate and n-hexane extracts, respectively. Ethyl acetate and n-hexane extracts exhibited strong antioxidant potential against 2,2-diphenyl-1-picrylhydrazil (DPPH) radical (IC50 values of 50.99 µg mL-1 and 74.44 µg mL-1, respectively). GC-MS analysis revealed 40 compounds in both extracts, some of which, including 2-ethylhexyl ester benzoic acid, benzo-b-dihydropyran-6-hydroxy-4-4-5-7-8-pentamethyl, diethyl phthalate, and octadecanoic acid, were identified through in silico analysis and found to possess antioxidant properties. These findings hold implications for potential applications of the plant and its biological constituent to be developed as lead compounds in the medical sector.

Production of codon-optimized Factor C fragment from Tachypleus gigas in the Pichia pastoris GS115 expression system for endotoxin detection.

BACKGROUND: Factor C (FC) is widely used as a standard material for endotoxin testing. It functions as a zymogenic serine protease and serve as a biosensor that detects lipopolysaccharides. Prior investigations involving molecular docking and molecular dynamics simulations of FC demonstrated an interaction between the C-type lectin domain (CLECT) and the ligand lipopolysaccharide (lipid A). In this study, our aim was to assess the stability of the interaction between fragment FC and the lipid A ligand using protein modeling approaches, molecular docking, molecular dynamics simulation, and gene construction into the pPIC9K expression vector. METHODS AND RESULTS: The FC structure was modelled by online tools. In this case, both molecular docking and MD simulations were applied to identify the interaction between protein and ligand (lipid A) including its complex stability. The FC structure model using three modeling websites has varied values, according to a Ramachandran plot study. When compared to other models, AlphaFold server modeling produced the best Ramachandran findings, with residues in the most advantageous area at 88.3%, followed by ERRAT values at 89.83% and 3D Verify at 71.93%. From the docking simulation of FC fragments with three ligands including diphosphoryl lipid A, FC-Core lipid A, and Kdo2 lipid A can be an activator of FC protein by binding to receptor regions to form ligand-receptor complexes. MD simulations were performed on all three complexes to assess their stability in water solvents showing that all complexes were stable during the simulation. The optimization of recombinant protein expression in Pichia pastoris was conducted by assessing the OD value and protease activity. Induction was carried out using 1% (v/v) methanol in BMMY media at 30°C for 72 h. CONCLUSIONS: Protein fragments of Factor C has been proven to detect endotoxins and serve as a potential biomarker. Molecular docking simulation and MD simulation were employed to study the complex formation of protein fragments FC with ligands. The expression of FC fragments was successfully achieved through heterologous expression. We propose optimizing the expression of FC fragments by inducing them with 1% methanol at 30°C and incubating them for 72 h. These optimized conditions are well-suited for upscaling the production of recombinant FC fragments using a bioreactor.

Properties and Application of Edible Modified Bacterial Cellulose Film Based Sago Liquid Waste as Food Packaging.

Bacterial cellulose (BC) based on sago liquid waste has been developed to be used as food packaging. This study investigated the physicochemical and mechanical properties of modified BC film and its application as food packaging. The modified BC film performed carboxymethyl cellulose (CMC) as a stabilizer and glycerol as a plasticizer. Films were prepared by casting technique using BC as the primary material and composites with various concentrations of CMC and glycerol (0.5%, 1%, and 1.5%, v/v). BC film was applied as the packaging of meat sausage, and the quality of meat sausage was measured based on weight loss, moisture content, pH, protein content, and total microbial count. The addition of CMC and glycerol influences the physical and mechanical properties of BC composites film. The best mechanical properties of edible BC film were collected by adding 1% CMC and 1% glycerol with a tensile strength of 17.47 MPa, elongation at a break of 25.60%, and Young's modulus of 6.54 GPa. FTIR analysis showed the characteristic bands of BC, and the addition of CMC and glycerol slightly changed the FTIR spectrum of the composites. The utilization of modified BC-based sago liquid waste film as the packaging of meat sausage could maintain sausage quality during 6 days of storage at room temperature. Therefore, edible BC film has the potential to be used as food packaging.

Isolation and enzyme bioprospection of bacteria associated to Bruguiera cylindrica, a mangrove plant of North Sumatra, Indonesia.

Mangrove-associated bacteria are of industrial interest due to their diverse and versatile enzyme properties. This study investigates the culturable bacteria from a wide range of habitat in a Bruguiera cylindrica mangrove ecosystem in North Sumatra. Screening of extracellular hydrolytic enzymes showed multiple potential traits in amylase, cellulase, chitinase, phosphatase, protease, and urease production by bacterial isolates. Molecular identification based on 16S rDNA region of a potential strain, Vibrio alginolyticus Jme3-20 is then reported as a newly proteolytic agent. The strain also showed a stable growth under salinity (NaCl) stress with considerable phosphate solubilization activities. Protease activity was enhanced by optimizing the 0.5 % (w/v) sucrose and soy peptone in the fermentation medium. SDS-PAGE and zymogram analysis showed the presence of a 35-kDa MW protease. Hence, our study revealed important insights into the bacterial diversity and activity in mangrove ecosystems, evidencing the importance of microbial exploration in this ecosystem.