In-vitro and in-silico analyses of the thrombolytic potential of green kiwifruit.

Cardiovascular diseases (CVDs), mainly caused by thrombosis complications, are the leading cause of mortality worldwide, making the development of alternative treatments highly desirable. In this study, the thrombolytic potential of green kiwifruit (Actinidia deliciosa cultivar Hayward) was assessed using in-vitro and in-silico approaches. The crude green kiwifruit extract demonstrated the ability to reduce blood clots significantly by 73.0 ± 1.12% (P < 0.01) within 6 h, with rapid degradation of Aα and Bß fibrin chains followed by the γ chain in fibrinolytic assays. Molecular docking revealed six favorable conformations for the kiwifruit enzyme actinidin (ADHact) and fibrin chains, supported by spontaneous binding energies and distances. Moreover, molecular dynamics simulation confirmed the binding stability of the complexes of these conformations, as indicated by the stable binding affinity, high number of hydrogen bonds, and consistent distances between the catalytic residue Cys25 of ADHact and the peptide bond. The better overall binding affinity of ADHact to fibrin chains Aα and Bß may contribute to their faster degradation, supporting the fibrinolytic results. In conclusion, this study demonstrated the thrombolytic potential of the green kiwifruit-derived enzyme and highlighted its potential role as a natural plant-based prophylactic and therapeutic agent for CVDs.

Genomic Characterization and Safety Assessment of Bifidobacterium breve BS2-PB3 as Functional Food.

Our group had isolated Bifidobacterium breve strain BS2-PB3 from human breast milk. In this study, we sequenced the whole genome of B. breve BS2-PB3, and with a focus on its safety profile, various probiotic characteristics (presence of antibiotic resistance genes, virulence factors, and mobile elements) were then determined through bioinformatic analyses. The antibiotic resistance profile of B. breve BS2-PB3 was also evaluated. The whole genome of B. breve BS2-PB3 consisted of 2,268,931 base pairs with a G-C content of 58.89% and 2,108 coding regions. The average nucleotide identity and whole-genome phylogenetic analyses supported the classification of B. breve BS2-PB3. According to our in silico assessment, B. breve BS2-PB3 possesses antioxidant and immunomodulation properties in addition to various genes related to the probiotic properties of heat, cold, and acid stress, bile tolerance, and adhesion. Antibiotic susceptibility was evaluated using the Kirby-Bauer disk-diffusion test, in which the minimum inhibitory concentrations for selected antibiotics were subsequently tested using the Epsilometer test. B. breve BS2-PB3 only exhibited selected resistance phenotypes, i.e., to mupirocin (minimum inhibitory concentration/MIC >1,024 µg/ml), sulfamethoxazole (MIC >1,024 µg/ml), and oxacillin (MIC >3 µg/ml). The resistance genes against those antibiotics, i.e., ileS, mupB, sul4, mecC and ramA, were detected within its genome as well. While no virulence factor was detected, four insertion sequences were identified within the genome but were located away from the identified antibiotic resistance genes. In conclusion, B. breve BS2-PB3 demonstrated a sufficient safety profile, making it a promising candidate for further development as a potential functional food.

Decolorization potential of malachite green by Ralstonia mannitolilytica isolated from Indonesian cassava-based fermented food tapai.

Pollution due to textile dye effluent mishandling is hazardous to ecosystems and to the living beings inhabiting them. This can cause retarded photosynthesis, disrupted fish day/night cycles, unbalanced bacterial populations, and decreased oxygen concentration in contaminated water, leading to low habitability. In this study, we aimed to isolate and characterize the microorganisms found in Indonesian cassava-based fermented food tapai starter cultures as a source of potential microbes for the biological remediation of textile dye pollutants. Microorganisms in the tapai starter culture were screened for their decolorization activity via spread-culture inoculation on a solid growth medium supplemented with textile dyes. Isolated microorganisms were selected based on their ability to secrete textile dye-decolorizing extracellular enzymes via increased light penetration after incubation of the cell-free supernatant (CFS) containing extracellular enzymes in textile dye solutions. Isolate JSP1 was the only bacterium capable of producing malachite green (MG)-decolorizing extracellular enzymes, which enabled it to survive and decolorize MG up to 375 ppm. Moreover, isolate JSP1 CFS was able to optimally decolorize 75% of MG at 100 ppm, but its activity was diminished at concentrations > 350 ppm. Colony and cellular morphology, biochemistry, and 16S rRNA tests revealed that the isolate was of Ralstonia mannitolilytica. Therefore, R. mannitolilytica isolate JSP1 may be a potential bioremediation agent for MG.