A b s t r a c tGamma irradiation is used on Penicillium cyclopium in order to obtain mutant cells of high L-asparaginase productivity. Using gamma irradiation dose of 4 KGy, P. cyclopium cells yielded L-asparaginase with extracellular enzyme activity of 210.8 ± 3 U/ml, and specific activity of 752.5 ± 1.5 U/mg protein, which are 1.75 and 1.53 times, respectively, the activity of the wild strain. The enzyme was partially purified by 40-60% acetone precipitation. L-asparaginase was immobilized onto Amberlite IR-120 by ionic binding. Both free and immobilized enzymes exhibited maximum activity at pH 8 and 40°C. The immobilization process improved the enzyme thermal stability significantly. The immobilized enzyme remained 100% active at temperatures up to 60°C, while the free asparaginase was less tolerant to high temperatures. The immobilized enzyme was more stable at pH 9.0 for 50 min, retaining 70% of its relative activity. The maximum reaction rate (V max ) and Michaelis-Menten constant (K m ) of the free form were significantly changed after immobilization. The K m value for immobilized L-asparaginase was about 1.3 times higher than that of free enzyme. The ions K + , Ba 2+ and Na + showed stimulatory effect on enzyme activity with percentages of 110%, 109% and 106% respectively. K e y w o r d s: Penicillium cyclopium, Amberlite IR-120, gamma irradiation, ionic binding immobilization, L-asparaginase

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The effect of oxygen on lipase production by Penicillium chrysogenum was studied under two operating modes, controlled aeration rate tested and controlled agitation at dissolved oxygen concentration (DO) 1.00 vvm. Lipase production and cell dry weight were tested in a stirred batch fermenter 5 L. Improvement in oxygen transfer rate (OTR) either by aeration or agitation resulted in an increase in lipase production. Growth curves and lipase activities of P.chrysogenum were examined at agitation rates (200,400,600 rpm), aeration rates (2,4 vvm) at different fermentation periods (24,48,72,96,120 h). Response Surface Methodology (RSM) using Design Expert software was used to study the effect of aeration, agitation, and fermentation time on lipase activity and cell dry weight. These factors were analyzed using 2 1 . 3 2 level factorial design. An optimal set of conditions that maximize lipase production: (2 vvm aeration; 600 rpm agitation after 72 h) was obtained. The maximum lipase activity obtained was 240 U/mL. Beside lipase activity, this paper also studies the optimal combination of the controllable factors (aeration; agitation and fermentation time) that will maximize the cell dry weight.

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