Optimization and reactor-scale production of plant growth regulators by Pleurotus eryngii.

The aims of the this study are to select the best cultivation type for plant growth regulator (PGR) production, to optimize PGR production with statistical experimental design, and to calculate bioprocess parameters and yield factors during PGR production by P. eryngii in flask and reactor scales. Submerged fermentation was the best cultivation type with 4438.67 ± 37.14, 436.95 ± 27.31, and 54.32 ± 3.21 mg/L of GA3, ABA, and IAA production values, respectively. The Plackett-Burman and Box-Behnken designs were used to determine effective culture parameters and interactive effects of the selected culture parameters on PGR production by Pleurotus eryngii under submerged fermentation. The statistical model is valid for predicting PGR production by P. eryngii. After these studies, maximum PGR production (7926.17 ± 334.09, 634.92 ± 12.15, and 55.41 ± 4.38 mg/L for GA3, ABA, and IAA, respectively) was reached on the 18th day of fermentation under optimized conditions. The optimum formula was 50 g/L fructose, 3 g/L NaNO3, and 1.5 g/L KH2PO4, 1 mg/L thiamine, incubation temperature 25 °C, initial medium pH 7.0, and an agitation speed of 150 rpm. The kinetics of PGR production was investigated in batch cultivation under 3-L stirred tank reactor conditions. Concentrations of GA3, ABA, and IAA of 10,545.00 ± 527.25, 872.32 ± 21.81, and 60.48 ± 3.48 mg/L were obtained at the reactor scale which were 4.1, 3.4, and 2.3 times higher than the initial screening values. The specific growth rate (µ), the volumetric (rp) and specific (Qp) PGR production rates, 486.11 mg/L/day and 107.43 mg/g biomass/day for GA3, confirmed the successful transfer of optimized conditions to the reactor scale. In the presented study, PGR production of P. eryngii is reported for the first time.

Flask and reactor scale production of plant growth regulators by Inonotus hispidus: optimization, immobilization and kinetic parameters.

The aims of the presented study are to compare submerged, static, and solid-state fermentation in the production of gibberellic acid (GA3), indole acetic acid (IAA), and abscisic acid (ABA) by Inonotus hispidus, to optimize with a statistical approach, and to determine the kinetic parameters under flask and reactor conditions. The maximum concentrations of GA3, (2478.85 ± 68.53 mg/L), ABA, (273.26 ± 6.17 mg/L) and IAA (30.67 ± 0.19 mg/L) were obtained in submerged conditions. After optimization, these values reached 2998.85 ± 28.85, 339.47 ± 5.50, and 34.56 ± 0.25 mg/L, respectively. Immobilization of fungal cells on synthetic fiber, polyurethane foam, and alginate beads resulted in an increase in plant growth regulators (PGR) production by 5.53%- 5.79% under optimized conditions. At the reactor scale, a significant increase was observed for GA3 concentration, 5441.54 mg/L, which was 2.14 and 1.45 times higher than non-optimized and optimized conditions in the flask scale, respectively. The maximum values for ABA and IAA were 390.39 and 44.79 mg/L, respectively. Although the specific growth rate (µ) decreases relatively from non-optimized flask conditions to optimized reactor conditions, it was observed that the PGR amounts produced per liter medium (rp) and per gram biomass (Qp) increased significantly. This is the first report on the synthesis of PGR by Inonotus hispidus which could be crucial for sustainable agriculture.

Optimization of Enzyme Co-Immobilization with Sodium Alginate and Glutaraldehyde-Activated Chitosan Beads.

In this study, two different materials-alginate and glutaraldehyde-activated chitosan beads-were used for the co-immobilization of α-amylase, protease, and pectinase. Firstly, optimization of multienzyme immobilization with Na alginate beads was carried out. Optimum Na alginate and CaCl2 concentration were found to be 2.5% and 0.1 M, respectively, and optimal enzyme loading ratio was determined as 2:1:0.02 for pectinase, protease, and α-amylase, respectively. Next, the immobilization of multiple enzymes on glutaraldehyde-activated chitosan beads was optimized (3% chitosan concentration, 0.25% glutaraldehyde with 3 h of activation and 3 h of coupling time). While co-immobilization was successfully performed with both materials, the specific activities of enzymes were found to be higher for the enzymes co-immobilized with glutaraldehyde-activated chitosan beads. In this process, glutaraldehyde was acting as a spacer arm. SEM and FTIR were used for the characterization of activated chitosan beads. Moreover, pectinase and α-amylase enzymes immobilized with chitosan beads were also found to have higher activity than their free forms. Three different enzymes were co-immobilized with these two materials for the first time in this study.

Anti-Quorum Sensing Potential of Antioxidant Quercetin and Resveratrol

ABSTRACT Quorum sensing system plays an active role in the regulation of pathogenicity of many microorganisms. Inhibition of pathogenicity or virulence factors will increase the success of treatment by preventing the development of antibiotic resistance. In this study, anti-quorum sensing activities of quercetin and resveratrol compounds, which have antioxidant property without damaging to host, have been determined via using biosensor bacteria: Chromobacterium violaceum ATCC 12472 and Chromobacterium violaceum CV026. As quorum sensing inhibitors, quercetin and resveratrol's cutting off the bacterial communication will prevent the treatment failures caused by the development of bacterial resistance. The development of layered drugs with antioxidant compounds such as quercetin and resveratrol will pave the way for new horizons for new therapeutic strategies.

Quorum sensing molecules production by nosocomial and soil isolates Acinetobacter baumannii.

Acinetobacter species remain alive in hospitals on various surfaces, both dry and moist, forming an important source of hospital infections. These bacteria are naturally resistant to many antibiotic classes. Although the role of the quorum sensing system in regulating the virulence factors of Acinetobacter species has not been fully elucidated, it has been reported that they play a role in bacterial biofilm formation. The biofilm formation helps them to survive under unfavorable growth conditions and antimicrobial treatments. It is based on the accumulation of bacterial communication signal molecules in the area. In this study, we compared the bacterial signal molecules of 50 nosocomial Acinetobacter baumannii strain and 20 A. baumannii strain isolated from soil. The signal molecules were detected by the biosensor bacteria (Chromobacterium violaceum 026, Agrobacterium tumefaciens A136, and Agrobacterium tumefaciens NTL1) and their separation was determined by thin-layer chromatography. As a result, it has been found that soil-borne isolates can produce 3-oxo-C8-AHL and C8-AHL, whereas nosocomial-derived isolates can produce long-chain signals such as C10-AHL, C12-AHL, C14-AHL and C16-AHL. According to these results, it is possible to understand that these signal molecules are found in the infection caused by A. baumannii. The inhibition of this signaling molecules in a communication could use to prevent multiple antibiotic resistance of these bacteria.

Escherichia Coli: Characteristics of Carbapenem Resistance and Virulence Factors

ABSTRACT In this study, fifty Escherichia coli strains were analyzed by multiplex polymerase chain reaction for the genes expressed carbapenemase and virulence factors in order to determine the presence of carbapenemase and nine virulence factors and investigate the association between these two characteristics. When carbapenemase susceptibility was taken into consideration, OXA-48 type carbapenemase was determined for 22% of the total strains. Also, the frequency of virulence gene regions in E.coli infections and virulence gene profiles of these isolates were examined and the frequency of pap, afa, sfa, fimA, iroN, aer, iutA, hly and cnf-1 genes were 24, 38, 20, 84, 28, 90, 92, 10 and 34% respectively. A significant correlation was found between the presence of fimA and afa gene regions and carbapenem susceptibility (P< 0.05). Based on the combination of carbapenemase and virulence factor genes, 24 different gene profiles were determined for all strains. The results of the study appear to indicate that fimA and afa genes correlate with carbapenem susceptibility, the relations of fimA with urinary tract infections and pap with complicated urinary tract infections. It also indicates that sfa and afa genes correlate with other infections except urinary tract infections.

Klebsiella pneumoniae: characteristics of carbapenem resistance and virulence factors.

Klebsiella pneumoniae, known as a major threat to public health, is the most common factor of nosocomial and community acquired infections. In this study, 50 K. pneumoniae clinical specimens isolated from bronchial, urea, blood, catheter, rectal, bile, tracheal and wound cultures were collected. These isolates were identified and carbapenem resistance was determined via an automated system, CHROMagar Orientation and CHROMagar KPC. The carbapenemase gene regions (blaIMP, blaVIM, blaOXA, blaNDM and blaKPC) and presence of virulence factors (magA, k2A, rmpA, wabG, uge, allS, entB, ycfM, kpn, wcaG, fimH, mrkD, iutA, iroN, hly ve cnf-1) of these isolates were determined by using Multiplex-PCR. The OXA-48 carbapenemase gene regions were determined in 33 of 50 K. pneumoniae strains. In addition, NDM-1 resistance in one, OXA-48 and NDM-1 resistance in four unusual K. pneumoniae isolates were detected. Virulence gene regions that were encountered among K. pneumoniae isolates were 88% wabG, 86% uge, 80% ycfM and 72% entB, related with capsule, capsule lipoprotein and external membrane protein, responsible for enterobactin production, respectively. Even though there was no significant difference between resistant and sensitive strains due to the virulence gene regions (P≥0.05), virulence factors in carbapenem resistant isolates were found to be more diverse. This study is important for both, to prevent the spread of carbapenem resistant infections and to plan for developing effective treatments. Moreover, this study is the first detailed study of the carbapenem resistance and virulence factors in K. pneumoniae strains.

Concanavalin A immobilized magnetic poly(glycidyl methacrylate) beads for prostate specific antigen binding.

The aim of this study was to prepare Concanavalin A (Con A) immobilized magnetic poly(glycidyl methacrylate) (mPGMA) beads for prostate specific antigen (PSA) binding and to study binding capacities of the beads using lectin-glycoprotein interactions. Firstly, iron oxide nanoparticles were synthesized by co-precipitation method and then, beads were synthesized by dispersion polymerization in the presence of iron oxide nanoparticles. Con A molecules were both covalently immobilized onto the beads directly and through the spacer arm (1,6-diaminohexane-HDMA). The total PSA and free PSA binding onto the mPGMA-HDMA-Con A beads were higher than that of the mPGMA-Con A beads. Maximum PSA binding capacity was observed as 91.2 ng/g. Approximately 45% of the bound PSA was eluted by using 0.1 M mannose as elution agent. The mPGMA-HDMA-Con A beads could be reused without a remarkable decrease in the binding capacities after 5 binding-desorption cycles. Serum fractions were analyzed using SDS-PAGE. The mPGMA-HDMA-Con A beads could be useful for the detection of PSA and suggested as a model system for other glycoprotein biomarkers.

Optimization of carbon-nitrogen ratio for production of gibberellic acid by Pseudomonas sp.

In this study, favorable carbon-nitrogen ratio for high yields of gibberellic acid (GA3) production from Pseudomonas sp. was investigated. First of all, optimum carbon (glucose, maltose, sucrose, fructose, lactose) and nitrogen (KNO3, NH4Cl, NaNO3, urea, glycine) sources among the others were chosen. The highest yield of GA3 productivity was found in growth medium supplemented with fructose (168.5 mg/L). NaNO3 was found as a suitable nitrogen source (141 mg/L). Then, in order to determine the optimum carbon-nitrogen ratio, different concentrations of carbon (from 50 mM to 150 mM) and nitrogen (from 17 mM to 47 mM) sources were added in culture media. As a result, optimum carbon-nitrogen ratio for GA3 production from Pseudomonas sp. was found to be 100:17 mM.