This study was carried out to evaluate the process of phytase production by Aspergillus niger in solidstate fermentation (SSF) using triticale waste. A waste that currently has no use was reported for this biotechnological process, and is of high impact due to the null use. The process was carried out using an additive free medium, supplemented with only one nitrogen source. Under these conditions, phytase activity of 7.45 U/g dry substrate (DS) was obtained. The process was optimized using different additives such as dextrose, lactose, Tween 80 and potassium chloride. For fermentation maximization, two experimental designs were used: 1) Plackett-Burman design (PBD) and 2) the Box-Behnken design (BBD). PBD was used to evaluate the effect of related variables on the production of phytase, as well as their level of significance in the process, while BBD was used for optimal conditions determination. The process was conducted with Petri dishes and a maximum enzyme activity of 25.8 IU/g DS was obtained. Subsequently, SSF was carried out in a tray to increase the amount of fermented substrate and phytase activity of 23.63 IU/g DS was obtained. The results of this study suggest a minimal decrease (8.4%) in enzyme production with scaling.

In order to seek an environmentally-friendly alternative to chemical fungicide, the aims of this paper were: 1) to assess the enzymatic and antifungal activity of free and microencapsulated enzymes (laminarinase and chitinase from Trichoderma sp.) and their mixtures against F. oxysporum in soil in the presence or absence of thiabendazole; 2) to analyze remarkable variants resulting from soil testing in ad planta assay with tomato crops (L. esculentum Mill) and the pathogen F. oxysporum. To evaluate the effect of applied treatments to the fungus, the quantity of colony forming units (CFU) was determined. Enzyme activities were measured spectrophotometrically using laminarin and of p-nitrophenyl-β-D-N-acetyl-glucosamide as laminarinase and chitinase substrates, respectively. An inhibiting effect to fungus growth (fungistatic effect) of both enzymes applied separately and together, as well as the treatment with liposomes only was demonstrated in vitro and ad planta assays. Enzyme application permitted a decreasing of chemical fungicide level and allowed the achievement of partial (90-98%) or total inhibition of fungus growth. Microencapsulation led to increase enzyme stability in the presence of thiabendazole. The growth of tomato plant was higher in the presence of applied treatments. The increase of fungistatic effect was revealed in the case of mixture of encapsulated enzymes and enzymes with thiabendazole. Encapsulation of chitinase and laminarinase in soya lecithin liposomes was useful to maintain enzymatic activity for the control of phytopathogenic fungus F. oxysporum. Thus, it confirmed the idea regarding the use of free or encapsulated (in soya lecithin liposome) mycolytic enzymes for the control of phytopathogen fungus growth in the presence and absence of chemical fungicide in vitro and ad planta assays.