화학공학소재연구정보센터
Journal of Supercritical Fluids, Vol.78, 143-148, 2013
Activity of cellulase and alpha-amylase from Hortaea werneckii after cell treatment with supercritical carbon dioxide
The aim of the research was to break the cell walls and membranes of Hortaea werneckii using SC CO2 and consecutively to release enzymes from cells with their unchanged activity after treatment. From the technical and economical reasons, microorganisms are still the most important source of enzymes. Therefore, the suspension culture of H. werneckii, which belongs to the black yeasts, was incubated in supercritical carbon dioxide (SC CO2) in order to use the enzymes from these fungi for biotransformations and compare their activity with the activity of purified commercial enzymes at the same conditions. Black yeasts contain many different enzymes; to cellular structures bound enzymes (intracellular enzymes), and extracellular enzymes. H. werneckii cell suspension was treated in SC CO2 at different pressure and fixed temperature (35 degrees C). The viability of H. werneckii cells was determined by measuring the optical density of the cell suspension at 600 nm. The total protein concentration and absorbance of nucleic acids in the suspension was determined on UV-Vis spectrophotometer at 595 nm and at 260 nm, respectively. The interest was to excrete the active intracellular enzyme alpha-amylase from H. werneckii and to keep the extracellular enzyme cellulase in the active form after the treatment of the cell suspension culture with SC CO2. The activity of intracellular enzyme eliminated from H. werneckii cells and that of extracellular enzyme after the treatment of the cells with SC CO2 was compared with the activity of alpha-amylase from Aspergillus oryzae in the powder form and cellulase from Trichoderma reesei, Cellusoft L, in the liquid form. (c) 2013 Elsevier B.V. All rights reserved.