Characterization, modeling, and anticancer activity of L.arginase production from marine Bacillus licheniformis OF2.

BACKGROUND: L-arginase, is a powerful anticancer that hydrolyzes L-arginine to L-ornithine and urea. This enzyme is widely distributed and expressed in organisms like plants, fungi, however very scarce from bacteria. Our study is based on isolating, purifying, and screening the marine bacteria that can produce arginase. RESULTS: The highest arginase producing bacteria will be identified by using microbiological and molecular biology methods as Bacillus licheniformis OF2. Characterization of arginase is the objective of this study. The activity of enzyme was screened, and estimated beside partial sequencing of arginase gene was analyzed. In silico homology modeling was applied to generate the protein's 3D structure, and COACH and COFACTOR were applied to determine the protein's binding sites and biological annotations based on the I-TASSER structure prediction. The purified enzyme was undergone an in vitro anticancer test. CONCLUSIONS: L-arginase demonstrated more strong anti-cancer cells with an IC50 of 21.4 ug/ml in a dose-dependent manner. L-arginase underwent another investigation for its impact on the caspase 7 and BCL2 family of proteins (BCL2, Bax, and Bax/Bcl2). Through cell arrest in the G1/S phase, L-arginase signals the apoptotic cascade, which is supported by a flow cytometry analysis of cell cycle phases.

Optimization of Bacillus subtilis growth parameters for biosynthesis of silver nanoparticles by using response surface methodology.

Silver nanoparticles (AgNPs) are among the most widely biosynthesized and used nanomaterials. They have different unique properties and a wide range of applications. This study is concerned with optimization of the growth conditions of Bacillus subtilis NRC1 for the biosynthesis of AgNPs using two designs of response surface methodology (RSM) statistical analysis. The data obtained from Plackett-Burman design (PBD) followed by central composite design (CCD), showed a good agreement between the experimental and predicted values of AgNPs biosynthesis. The optimum conditions were 0.7% (w/v) casein hydrolysate, 5% dextrin (w/v), pH 7.5 and 57 × 106 CFU/ml inoculum size. The model was highly valid and could be applied with a confidence factor of 99.47%. Minimal inhibitory concentration (MIC) of these AgNPs synthesized using the extracellular filtrate after growth of Bacillus subtilis NRC1 in the optimized medium was found to be 41-43µg/ml for all tested microorganisms with exception of Pseudomonas aeruginosa where MIC was 169 µg/ml.

Production and characterization of polyhydroxybutyrate bioplastic precursor from Parageobacillus toebii using low-cost substrates and its potential antiviral activity.

There is an increasing desire for bioplastics produced from renewable resources as an alternative to their petrochemical counterparts. These biopolymers have long-unnoticed antiviral properties. This study aimed to produce and characterize bioplastics by Parageobacillus toebii using low-cost substrates and determine their antiviral activity against coxsackievirus B4. Seven low-cost substrates (bagasse, water hyacinth, rice straw, rice water, sesame husks, molasses, and corn syrup) were compared with glucose for bioplastic precursor production. The highest bioplastic produced was from water hyacinth and glucose, followed by molasses, rice straw, rice water, sesame husks, and bagasse. Water hyacinth and glucose media were further optimized to increase the bioplastic precursor yield. The optimization of the media leads to increases in bioplastic precursor yields of 1.8-fold (3.456 g/L) and 1.496-fold (2.768 g/L), respectively. These bioplastics were further characterized by thermogravimetric analysis (TGA), Fourier-transformed infrared (FTIR) spectroscopy, proton nuclear magnetic resonance (1H NMR), and gas chromatography-mass spectrometry (GC-MS). They are thermostable, and their characterizations confirm the presence of polyhydroxybutyrate. The antiviral assay showed reasonable antiviral effects for bioplastics from water hyacinth (80.33 %) and glucose (55.47 %) media at 250 µg/mL maximum non-toxic concentrations (MNTC). The present investigation demonstrates a low-cost model for producing polyhydroxybutyrate bioplastic precursor for antiviral applications.

Insight into antioxidant and anti-inflammatory effects of marine bacterial natural exopolysaccharide (EPSSM) using carrageenan-induced paw edema in rats.

Inflammation is a part of the body's intricate biological reaction to noxious stimuli and defensive reactions. So, the aim of this investigation was to study the anti-inflammatory activity of exopolysaccharide (EPSSM) using carrageenan-induced paw edema in rats. A halophilic bacterial strain was isolated from marine sediments in the Red Sea in Egypt. The isolate has been visually and physiologically recognized, as well as by analyzing its 16S rRNA gene, which confirms Kocuria sp. clone Asker4. This particular isolate can be referenced using the accession number OL798051.1. EPSSM was subjected to purification and fractionation by a DEAE-cellulose column. Preliminary chemical analysis of EPSSM indicated that the monosaccharides were fructose, glucuronic acid, and xylose, with 2.0, 0.5, and 1.0, respectively. The antioxidant potential of EPSSM was investigated, and it was discovered that the level of activity increased independently of the concentrations, reaching a maximum threshold of 94.13% at 100 µg/mL of EPSSM for 120 min. Also, EPSSM at 50 mg/kg orally produced a significant anti-inflammatory effect on the carrageenan model at 2, 3, and 4 intervals. The EPSSM intervention resulted in reductions in the levels of catalase and superoxide dismutase enzymes, as well as a decrease in glutathione. Furthermore, the levels of nitric oxide, lipid peroxidation, and reactive oxygen species resulting from carrageenan-induced edema showed a significant reduction subsequent to the administration of EPSSM. Moreover, the findings indicated that the protein expression levels of cyclooxygenase-2 and interleukin-6 were reduced following treatment with EPSSM, resulting in a reduction of paw edema.

Modeling and in-vivo evaluation of fibrinolytic enzyme produced by Bacillus subtilis Egy under solid state fermentation.

Blood clot formation increases cases of myocardial infarction (AMI) and stroke, thus urges directing much research works for treatment and prevention of the causes. One of these directions is the microbial production of fibrinolytic enzymes as thrombolytic agents. In the current work, Bacillus subtilis Egy has been used for enzyme production under solid state fermentation. Among twelve nutrient meals in addition to wheat bran as a control fodder yeast yielded the highest enzyme activity reaching 114U/g. Applying statistical model for optimization of enzyme production revealed that 3.6%, fodder yeast; 40%, moisture content; 6 days, incubation period and 2%, inoculum size were the optimum conditions for maximum fibrinolytic enzyme production (141.02 U/g) by Bacillus subtilis Egy under solid-state fermentation The model was significant and data were experimentally validated. The produced fibrinolytic enzyme was evaluated for in vitro and in vivo cytotoxicity. In-vivo examination of the enzyme resulted in no mortality during the first 24 h after treatment. After 14 days, the results revealed no significant changes detected in hematological parameters (RBCs, MCV, hemoglobin except WBCs which showed an increase for both sexes. Histopathological examination of liver and kidney of rats received oral and subcutaneous treatments showed normal architecture. The data showed the applicability of the produced enzyme for the treatment of blood clot with no significant effect on living cells or on physiological functions.

Synthesis and investigations on tellurium myconanoparticles.

Tellurium has attracted the attention of many researchers and manufacturers due to its unique properties. Through the current work, six fungal isolates have been screened for their ability to reduce potassium tellurite (K2TeO3) into elemental tellurium nanoparticles (TeNPs). The most promising fungal isolate was identified as Aspergillus welwitschiae and given the accession number (KY766958) based on molecular basis and has been used for biogenic (enzymatic) production of TeNPs. The produced TeNPs have been characterized using DLS, TEM and FTIR. Data showed that, the particle size is 60.80 d.nm with oval to spherical shape. The produced TeNPs have been evaluated for antimicrobial activity at 25 mg/ml. Data revealed antibacterial activity against E. coli and Staphylococcus aureus (MRSA). Evaluation of the effect of γ-irradiation on TeNPs production showed that, the productivity was improved at 1 kGy and suppressed gradually at higher doses.

Optimization of biosurfactant production by Bacillus brevis using response surface methodology.

The present study aims to evaluate and validate a statistical model for maximizing biosurfactant productivity by Bacillus brevis using response surface methodology. In this respect, twenty bacterial isolates were screened for biosurfactant production using hemolytic activity, oil spreading technique, and emulsification index (E24). The most potent biosurfactant-producing bacterium (B. brevis) was used for construction of the statistical response surface model. The optimum conditions for biosurfactant production by B. brevis were: 33 °C incubation temperature at pH 8 for 10 days incubation period and 8.5 g/L glucose concentration as a sole carbon source. The produced biosurfactant (BS) (73%) exhibited foaming activity, thermal stability in the range 30-80 °C for 30 min., pH stability, from 4 to 9 and antimicrobial activity against (Escherichia coli). The BS gave a good potential application as an emulsifier.

Optimization of lovastatin production from Aspergillus fumigatus.

The present investigation, focused on screening of various fungal species for Lovastatin production using different agro-based wastes, also, for maximizing lovastatin productivity by isolated Aspergillus fumigatus using response surface methodology (RSM). The following substrates (Olive cake; Pea pods; sugarcane bagasse; wheat bran; rice hulls; beet peel; Potato peel and groundnut shells) were screened to evaluate their effectiveness for lovastatin production, using different fungal species, (Aspergillus niger; Rhizopus oligosporus; Penicillium citrinum and isolated Aspergillus fumigatus) under solid state fermentation (SSF). Wheat bran was the most suitable substrate for lovastatin production with all fungal species. Optimum conditions of lovastatin production by wheat bran have been attained efficiently by response surface methodology (RSM) using isolated Aspergillus fumigatus under solid state fermentation (SSF). The lovastatin yield of (3.353 mg/g DFM) was obtained at an optimum temperature of 28 °C; pH of 5.00; initial moisture content of 70% and incubation period of 12 days. This Lovastatin has the possibility to use in different therapeutic applications.