Physiological and biochemical studies of microbiological production of chitosanase /
Shaymaa Abdalla Ismail
Physiological and biochemical studies of microbiological production of chitosanase / دراسات فسيولوجيه و بيوكيميائيه على انتاج انزيم الكيتوزانيز ميكروبيا Shaymaa Abdalla Ismail ; Supervised Alaa Eldein M. S. Hosny , Amal Mohamed Hashem - Cairo : Shaymaa Abdalla Ismail , 2016 - 182 P. : charts , photographs ; 25cm
Thesis (Ph.D.) - Cairo University - Faculty of Pharmacy - Department of Microbiology and Immunology
Over 13 microorganisms screened for chitosanase production, an isolate identified as Dothideomycetes sp. css035 secreted a considerable value of inducible chitosanase (1.27 U/ml). The optimization of the medium components and the culture conditions was performed by using one factor at a time technique followed by statistical methods including plackett-burman design and Box-Behnken design. The highest chitosanase production was induced by the medium composition g/L: soluble chitosan, 30; K₂HPO₄, 1.15; MgSO₄, 0.4; KCl, 4.0; yeast extract, 18.5; FeSO₄, 0.01; at pH 5.5, 30 C and 180 rpm for four days. The optimization study was markedly enhancing the production of the extracellular and the cell bound chitosanase from 1.27 U/ml and 11.9 U/ dry weight to 13.9 U/ml and 215.83 U/ dry weight. The crude extracellular chitosanase was optimally active (21.5 U/ml) at pH 4.5 and temperature 60C with 1% soluble chitosan and it exhibited a good stability over pH range from 4 to 5.5 and thermostability below 45C in the absence of the chitosan since the enzyme had half life time higher than 2 h. The immobilization of 30-70% ethanol precipitated Dothideomycetes sp. css035 chitosanase by using covalent binding to carrageenan: carboxymethyl cellulose carrier that showed improved pH and thermal stability in comparison to the partial pure enzyme. Thin-layer chromatography plate (TLC) clearly visualized the efficiency of the produced chitosanase to hydrolyze chitosan to chitooligosaccharides and the highest concentration of chitooligosaccharides was produced after 5 h reaction time at 50 C using 2% chitosan and E/S ratio 0.05 U/mg. The biological activities evaluation of the produced chitooligosaccharides mixture (eluted from the TLC) indicated that the chitooligosaccharides mixture showed fibrinolytic and anticancer activity
Chitosanase Microbiological production Microbiology and Immunology
Physiological and biochemical studies of microbiological production of chitosanase / دراسات فسيولوجيه و بيوكيميائيه على انتاج انزيم الكيتوزانيز ميكروبيا Shaymaa Abdalla Ismail ; Supervised Alaa Eldein M. S. Hosny , Amal Mohamed Hashem - Cairo : Shaymaa Abdalla Ismail , 2016 - 182 P. : charts , photographs ; 25cm
Thesis (Ph.D.) - Cairo University - Faculty of Pharmacy - Department of Microbiology and Immunology
Over 13 microorganisms screened for chitosanase production, an isolate identified as Dothideomycetes sp. css035 secreted a considerable value of inducible chitosanase (1.27 U/ml). The optimization of the medium components and the culture conditions was performed by using one factor at a time technique followed by statistical methods including plackett-burman design and Box-Behnken design. The highest chitosanase production was induced by the medium composition g/L: soluble chitosan, 30; K₂HPO₄, 1.15; MgSO₄, 0.4; KCl, 4.0; yeast extract, 18.5; FeSO₄, 0.01; at pH 5.5, 30 C and 180 rpm for four days. The optimization study was markedly enhancing the production of the extracellular and the cell bound chitosanase from 1.27 U/ml and 11.9 U/ dry weight to 13.9 U/ml and 215.83 U/ dry weight. The crude extracellular chitosanase was optimally active (21.5 U/ml) at pH 4.5 and temperature 60C with 1% soluble chitosan and it exhibited a good stability over pH range from 4 to 5.5 and thermostability below 45C in the absence of the chitosan since the enzyme had half life time higher than 2 h. The immobilization of 30-70% ethanol precipitated Dothideomycetes sp. css035 chitosanase by using covalent binding to carrageenan: carboxymethyl cellulose carrier that showed improved pH and thermal stability in comparison to the partial pure enzyme. Thin-layer chromatography plate (TLC) clearly visualized the efficiency of the produced chitosanase to hydrolyze chitosan to chitooligosaccharides and the highest concentration of chitooligosaccharides was produced after 5 h reaction time at 50 C using 2% chitosan and E/S ratio 0.05 U/mg. The biological activities evaluation of the produced chitooligosaccharides mixture (eluted from the TLC) indicated that the chitooligosaccharides mixture showed fibrinolytic and anticancer activity
Chitosanase Microbiological production Microbiology and Immunology