Production, bioprocess optimization and γ-irradiation of Penicillium polonicum, as a new Taxol producing endophyte from Ginko biloba.

Twenty-eight fungal endophytes were recovered from the different parts of Ginkgo biloba and screened for their Taxol producing potency. Among these isolates, Penicillium polonicum AUMC14487 was reported as the potent Taxol producer (90.53 µg/l). The chemical identity of the extracted Taxol was verified from the TLC, HPLC, NMR, EDX, and FTIR analyses. The extracted Taxol displayed a strong antiproliferative activity against HEPG2 (IC50 4.06 µM) and MCF7 (IC50 6.07 µM). The yield of Taxol by P. polonicum was optimized by nutritional optimization with the Response Surface Methodology (RSM) using Plackett-Burman and Central Composite Designs. In addition to nutritional optimization, the effect of γ-irradiation of the spores of P. polonicum on its Taxol producing potency was evaluated. The yield of Taxol by P. polonicum was increased via nutritional optimization by response surface methodology with Plackett-Burman and FCCD designs, and γ-irradiation by about 4.5 folds, comparing to the control culture. The yield of Taxol was increased by about 1.2 folds (401.2 µg/l) by γ -irradiation of the isolates at 0.5-0.75 kGy, comparing to the control cultures (332.2 µg/l). The highest Taxol yield was obtained by growing P. polonicum on modified Czapek's- Dox medium (sucrose 40.0 g/l, malt extract 20.0 g/l, peptone 2.0 g/l, K2PO4 2.0 g/l, KCl 1.0 g/l, NaNO3 2.0 g/l, MgSO4. 5H2O 1.0 g/l) of pH 7.0 at 30.0 °C for 7.0 days. From the FCCD design, sucrose, malt extract and incubation time being the highest significant variables medium components affecting the Taxol production by P. polonicum.

XRD and ATR-FTIR techniques for integrity assessment of gamma radiation sterilized cortical bone pretreated by antioxidants.

Terminal sterilization of bone allograft by gamma radiation is required to reduce the risk of infection. Free radical scavengers could be utilized to minimize the deteriorating effects of gamma radiation on bone allograft mechanical properties. The objective of this research is to assess the changes in structural and chemical composition induced by hydroxytyrosol (HT) and alpha lipoic acid (ALA) free radical scavengers in gamma sterilized cortical bone. Bovine femurs specimens were soaked in different concentrations of HT and ALA for 7 and 3 days respectively before irradiation with 35 KGy gamma radiation. The attenuated total reflection-Fourier transform infrared spectroscopy and the X-ray diffraction techniques were utilized to analyze the changes in chemical composition induced by irradiation in the presence of free radical scavengers. A significant increase in the proportion of amide I and amide II to phosphate was noticed in the irradiated group, while in the pretreated groups with ALA and HT this effect was minimized. In addition, gamma radiation reduced the mature to immature cross links while ALA and HT alleviated this reduction. No significant changes were noticed in the mineral crystallinity or crystal size. Bone chemical structure has been changed due to gamma irradiation and these changes are mainly relevant to amide I, amide II proportions and collagen crosslinks. The deteriorating effects of gamma sterilization dose (35 kGy) on chemical structure of bone allograft can be alleviated by using (HT) and (ALA) free radical scavengers before irradiation.

Biomechanical properties enhancement of gamma radiation-sterilized cortical bone using antioxidants.

Gamma radiation sterilization is the method used by the majority of tissue banks to reduce disease transmission from infected donors to recipients through bone allografts. However, many studies have reported that gamma radiation impairs the structural and mechanical properties of bone via formation of free radicals, the effect of which could be reduced using free radical scavengers. The aim of this study is to examine the radioprotective role of hydroxytyrosol (HT) and alpha lipoic acid (ALA) on the mechanical properties of gamma-sterilized cortical bone of bovine femur, using three-point bending and microhardness tests. Specimens of bovine femurs were soaked in ALA and HT for 3 and 7 days, respectively, before being exposed to 35-kGy gamma radiation. In unirradiated samples, both HT and ALA pre-treatment improved the cortical bone bending plastic properties (maximum bending stress, maximum bending strain, and toughness) without affecting microhardness. Irradiation resulted in a drastic reduction of the plastic properties and an increased microhardness. ALA treatment before irradiation alleviated the aforementioned reductions in maximum bending stress, maximum bending strain, and toughness. In addition, under ALA treatment, the microhardness was not increased after irradiation. For HT treatment, similar effects were found. In conclusion, the results indicate that HT and ALA can be used before irradiation to enhance the mechanical properties of gamma-sterilized bone allografts.

Biosynthesis of gibberellic acid from milk permeate in repeated batch operation by a mutant Fusarium moniliforme cells immobilized on loofa sponge.

Gibberellic acid (GA) production from milk permeate was studied by 28 mutants of Fusarium moniliforme, among which mutant gamma-14 was selected as the best producer. Experiments were carried out in shaker flasks and fermentative process was analyzed with free and immobilized cells. Immobilization of mutant gamma-14 cells onto loofa sponge discs was studied with respect to the optimization of the incubation temperature, initial pH, inoculum size (number of discs) and its reusability for GA production. Best yield of GA (2.40 gl(-1)) was recorded by immobilized cells under optimized cultural conditions (4 immobilized discs, 30 degrees C and pH 5). Data obtained during four reusable cycles showed high stability of GA production and reduction in the initiation time of acid production, resulting in higher levels of GA in shorter time duration. Immobilization of mutant gamma-14 cells onto loofa sponge discs, permitted repeated reuse under the specified fermentation conditions for GA production from milk permeate.

Biosorption of reactive dye from textile wastewater by non-viable biomass of Aspergillus niger and Spirogyra sp.

The potential of Aspergillus niger fungus and Spirogyra sp., a fresh water green algae, was investigated as a biosorbents for removal of reactive dye (Synazol) from its multi component textile wastewater. The results showed that pre-treatment of fungal and algal biomasses with autoclaving increased the removal of dye than pre-treatment with gamma-irradiation. The effects of operational parameters (pH, temperature, biomass concentration and time) on dye removal were examined. The results obtained revealed that dried autoclaved biomass of A. niger and Spirogyra sp. exhibited maximum dye removal (88% and 85%, respectively) at pH3, temperature 30 degrees C and 8 gl(-1)(w/v) biomass conc. after 18h contact time. The stability and efficiency of both organisms in the long-term repetitive operation were also investigated. The results showed that the non-viable biomasses possessed high stability and efficiency of dye removal over 3 repeated batches.