18 research outputs found
Kinetics of Hypoglycemic α-Glucosidase Inhibitory Protein
The activity of a-glucosidase enzyme (EC 3.2.1.20) was inhibited by 40.3% by an extracellular inhibitory protein which isolated from the culture filtrate of Aspergillus brasiliensis ATCC-16404. The maximum activity of this protein and fungal mycelial dry weight were obtained at 28°C, 7 days, 6.5, 400 µl/ml, 140 rpm, and rice straw as the optimum incubation temperature, incubation period, pH value, inoculum size, agitation speed, and raw material respectively. The a-glucosidase inhibitory protein (AGIP) was purified and separated electrically as a single band at 40 KDa. There 17 amino acids of the purified protein were determined at different concentrations, where threonine has a highest percentage (90%) and aspartic acid has a lower percentage (24%). Kinetics of AGIP were determined, where Vmax, Km, kcat, and catalytic activity values were exactly calculated using Lineweaver-Burk plot compared with those of acarbose as an inhibitor standard. The maximum catalytic activity (7.808 M-1S-1) at 0.1 mg/mol was higher than that of acarbose (7.783 M-1S-1) at the same concentration
Dissolution of clotted blood from slaughtered cattle by alkaline protease produced by Streptomyces violaceoruber isolated from saline soil in Saudi Arabia
Saudi Arabia suffers from the spread of diseases linked to clotted blood leaking from slaughtered livestock. Anticoagulant proteases produced by Streptomyces are used to safely dissolve clotted blood. Halophilic Streptomyces isolates were extracted from saline soil in Saudi Arabia and tested to dissolve clotted blood by protease. The most protease-producing isolate was identified using classical and genetic methods. The protease was precipitated with saturated ammonium sulfate from a batch of blood clots inoculated with the most protease-producing isolates. Streptomyces isolates (410) were recovered from thirty saline soil samples collected from Dammam (170 isolates, 41.4 %), Al-Khobar (126 isolates, 30.8 %) and Al-Ahsa (114 isolates, 27.8 %). Proteolytic activity was observed in 305 isolates including the most protease-producing isolate S295, which showed the highest activity (33.8 mm), and was identified as S. violaceoruber (99 % similarity). Among all isolates, 241 showed partial hemolysis (alpha) and 64 showed complete hemolysis (beta), including isolate S295 that presented the highest hemolytic activity (0.58 %). The alkaline protease was precipitated with saturated ammonium sulfate (60 %), and it was found that its activity was 5.2 U/ml, total activity 24,960 U/ml, specific activity 463 U/mg, purification fold 15.2 and yield 78 %. The purified alkaline protease was electrophoresed as a single band at 45 kDa
Enhancement of fungal DNA templates and PCR amplification yield by three types of nanoparticles
Nanodiagonastic methods in plant pathology are used for enhancing detection and identification
of different plant pathogens and toxigenic fungi. Improvement of the specificity and
efficiency of the polymerase chain reaction (PCR) by using some nanoparticles is emerging
as a new area of research. In the current research, silver, zinc, and gold nanoparticles
were used to increase the yield of DNA for two plant pathogenic fungi including soil-borne
fungus Rhizoctonia solani and toxigenic fungus Alternaria alternata. Gold nanoparticles
combined with zinc and silver nanoparticles enhanced both DNA yield and PCR products
compared to DNA extraction methods with ALB buffer, sodium dodecyl sulfate, ALBfree
from protinase K, ZnNPs and AgNPs. Also, by using ZnNPs and AgNPs the DNA yield
was enhanced and the sensitivity of random amplified polymorphic DNA (RAPD) PCR
products was increased. Application of nanomaterials in the PCR reaction could increase
or decrease the PCR product according to the type of applied nanometal and the type of
DNA template. Additions of AuNPs to PCR mix increased both sensitivity and specificity
for PCR products of the tested fungi. Thus, the use of these highly stable, commercially
available and inexpensive inorganic nano reagents open new opportunities for improving
the specificity and sensitivity of PCR amplicon, which is the most important standard
method in molecular plant pathology and mycotoxicology
Studies on wound healing potential of red pigment isolated from marine Bacterium Vibrio sp.
Wounds are common clinical entities of life which may be subacute or acute. Wound healing is a complex biochemical process where the cell structures are restored to normalcy, which depend on cell proliferation and migration, basically fibroblast cell. The present investigation was undertaken to evaluate the healing efficacy of red pigment isolated from marine isolate Vibrio sps on experimental wounds in albino rats. The red pigment was applied topically, twice daily for 14 days. Treatment with framycetin ointment was used as reference control. The red pigment treated group showed faster reduction in wound area in comparison with control and framycetin ointment treated groups. In conclusion, red pigment possesses significant healing potential in wounds and has a positive influence on the different phases of wound repair. Keywords: Pigments, Marine microbes, Vibrio sps, Wound healing, Anti bacteria