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[en] The effect of different concentrations of cellulase on the production of capsaicin in freely suspended cell and immobilized cell cultures of Kahramanmara pepper seeds (Capsicum annuum L.) were studied. Calluses were obtained from in vitro germinated hypocotyl explants of pepper seedlings and cell suspensions were prepared from these calluses. Immobilized cell suspension cultures with calcium alginate and free cell suspension cultures were obtained by using cell suspensions. Elicitor such as cellulase (5-30 micro g/ml), was applied both for the free and immobilized cell suspensions and control group without elicitor was prepared. The concentration of capsaicin in freely suspended cells, immobilized cells and their filtrates were identified by HPLC after extraction with ethyl acetate. It was found that the immobilization process had an increasing effect on the capsaicin accumulation. The concentration of capsaicin in the immobilized cells for both control groups and elicitor added samples was higher than the free cells. In general, capsaicin concentration in the filtrate for free cells was higher than the immobilized cells. When all the cellulase and the sampling hours were compared, the highest capsaicin concentration for the immobilized cells was determined as 362,91 micro g/ml f.w. at the 24th hour for 30 micro g/ml cellulase applied samples. (author)
[en] The dependence of the Moessbauer elastic scattering fraction on the hydration degree (h) has been studied for hydrated samples of human albumen (HSA), lysozyme and trypsin pancreatic inhibitor, within the range of h between 0-0.75 g H2O/g protein at 295 K, and for HSA with different hydration degrees (0.03, 0.25, 0.41, 0.65) in the temperature range between 100-320 K. An increase of the hydration degree for h>0.1 at T>200 K has been shown to result in the release of intramolecular mobility in proteins. (orig.)
[en] Thermo stable cellulase from fungi has high potential for industrial application. In this study, wild -type of fungal were isolate from different sources such as hot spring water, sea water, soft wood, rice straw and cow dung. The isolates were characterized by cultural and morphological observation. Based on morphological characteristics, the genera of all fungal cultures were identified namely Aspergillus fumigatus. The screening for thermo stable cellulase were done using 2 % carboxymethyl cellulose and congo red as an indicator at temperature 30, 37, 45 and 50 degree Celsius respectively. Out of 26 fungal isolates, only eight isolates were selected for further screening and showed the abilities to secrete cellulases by forming distinct halo zones on selective agar plate. The maximum halo zone ranging from 32 mm to 35 mm were obtained after 72 hour incubation at 50 degree Celsius by H2, SW1 and C1 isolates. As contrary other isolates showed halo zone range from 22 mm to 29 mm at same temperature. All the isolates showed the abilities to secrete cellulase enzyme at other temperature but lower when compared to 50 degree Celsius referred to the halo zone obtained. The SW1 isolates showed highest cellulolytic index which was 2.93 measured at 37 degree Celsius and 2.67 at 50 degree Celsius respectively. (author)
[en] Autophagy is a lysosome-dependent catabolic process involving in the degradation and recycling of unnecessary or damaged proteins and organelles. Emerging evidence indicates that autophagy dysfunction is closely related to various human diseases including cancer, aging, myopathies and neurodegenerative disorders. Here, using genetic knockdown, we uncover the role of Numb, an endocytic adaptor protein, in regulating the late steps of autophagy. We found that Numb depletion led to the accumulation of autophagic vacuole, as verified by RFP-LC3 staining combined with transmission electron microscopy. Further investigation indicated that Numb depletion impaired autophagic degradation through inhibiting the activities of lysosomal enzymes (Cathepsin D, β-glucuronidase and β-glucosidase). Moreover, Numb depletion induced elevation of lysosomal pH values and decrease of glycosylated lysosome-associated membrane proteins. We further observed that Rab7 activity was inhibited in Numb-depleted cells. Together, our findings revealed a novel function of Numb and its likely mechanism in regulation of autophagy events. - Highlights: • Numb depletion induces the accumulation of autophagic vacuole in MCF-7 cells. • Numb depletion impairs autophagic degradation and lysosomal enzymes activities. • Numb depletion elevates lysosomal pH but decreases glycosylated LAMPs. • Numb may regulate lysosome maturation by facilitating Rab7 activation.
[en] In this study traditional microbiological isolation techniques were used to recognize cellulolytic activity of most potent isolates. The isolates were identified using both morphological and molecular methods. Functional studies were carried out to determine the optimum pH, temperature, incubation period, inoculum size, carbon and nitrogen sources for screened isolate. High cellulase producing endo phytic bacteria Achromobacter spanius, Bacillus amyloliquefaciens and Stenotrophomonas maltophilia isolated from leaves, stems and roots of; Clover, Wheat, Lettuce, Spinach, Vicia Faba, Garlic, Dill, Olive, Acacia and Prosopis trees were subjected to controlled conditions in laboratory. Tested factors which believed to be effective on improving growth conditions were studied in vitro to achieve the highest production of cellulase enzyme. The highest values of cellulase were produced by Achromobacter spanius (2.50 mg/ml) followed by Bacillus amyloliquefaciens (2.26 mg/ml) after 6 days of incubation. The optimum conditions resulted in optimum cellulase production were 30°C at ph 6 for Achromobacter and Bacillus while the optimum conditions for Stenotrophomonas was 40°C at ph 7. Isolates produced the highest values of cellulase with ammonium sulphate and 30 g/l glucose as nitrogen and carbon sources, respectively. Achromobacter was more effective in producing cellulase than Bacillus and Stenotrophomonas. This was true under all the tested conditions.