Permeabilization was used for the purpose of transforming the cells of microorganisms into biocatalysts with an enhanced enzyme activity. Baker’s yeast cells were permeabilized with various organic solvents. A high degree of catalase activity was observed upon permeabilization with acetone, chloroform, isopropyl alcohol and ethyl acetate. Response surface methodology was used to model the effect of concentration of isopropyl alcohol, temperature and treatment time on the permeabilization of baker’s yeast cells to maximize the decomposition of H2O2. The optimum operating conditions for permeabilization were observed at 53.7% concentration of isopropyl alcohol, treatment time of 40 min and temperature of 15.6oC. A maximum value of catalase activity was found to be 6.188 U/g wet wt. and was ca. 60 times higher than the catalytic activity of yeast not treated by the permeabilization process.
Permeabilization is one of the effective tools, used to increase the accessibility of intracellular enzymes. Immobilization is one of the best approaches to reuse the enzyme. Present investigation use both techniques to obtain a biocatalyst with high catalase activity. At the beginning the isopropyl alcohol was used to permeabilize cells of baker’s yeast in order to maximize the catalase activity within the treated cells. Afterwards the permeabilized cells were immobilized in calcium alginate beads and this biocatalyst was used for the degradation of hydrogen peroxide to oxygen and water. The optimal sodium alginate concentration and cell mass concentration for immobilization process were determined. The temperature and pH for maximum decomposition of hydrogen peroxide were assigned and are 20°C and 7 respectively. Prepared biocatalyst allowed 3.35-times faster decomposition as compared to alginate beads with non permeabilized cells. The immobilized biocatalyst lost ca. 30% activity after ten cycles of repeated use in batch operations. Each cycles duration was 10 minutes. Permeabilization and subsequent immobilization of the yeast cells allowed them to be transformed into biocatalysts with an enhanced catalase activity, which can be successfully used to decompose hydrogen peroxide.
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Saccharamyces cerevisia known as baker’s yeast is a product used in various food industries. Worldwide economic competition makes it a necessity that industrial processes be operated in optimum conditions, thus maximisation of biomass in production of saccharamyces cerevisia in fedbatch reactors has gained importance. The facts that the dynamic fermentation model must be considered as a constraint in the optimisation problem, and dynamics involved are complicated, make optimisation of fed-batch processes more difficult. In this work, the amount of biomass in the production of baker’s yeast in fed-batch fermenters was intended to be maximised while minimising unwanted alcohol formation, by regulating substrate and air feed rates. This multiobjective problem has been tackled earlier only from the point of view of finding optimum substrate rate, but no account of air feed rate profiles has been provided. Control vector parameterisation approach was applied the original dynamic optimisation problem which was converted into a NLP problem. Then SQP was used for solving the dynamic optimisation problem. The results demonstrate that optimum substrate and air feeding profiles can be obtained by the proposed optimisation algorithm to achieve the two conflicting goals of maximising biomass and minimising alcohol formation.
Przedstawiono wyniki badań procesu permeabilizacji komórek drożdży piekarskich. Doświadczenia prowadzono zgodnie z planem rotatabilnym rzędu drugiego. Na ich podstawie sformułowano model matematyczny, który pozwolił wyznaczyć optymalne warunki przebiegu procesu. Komórki drożdży traktowano przez 103 min 37,7-proc. roztworem metanolu w temp. 21,4°C. Stała szybkości rozkładu H₂O₂ dla drożdży permeabilizowanych była 9 razy większa niż dla drożdży natywnych.
EN
Baker’s yeast cells were permeabilized to det. the optimal conditions for the process. The yeast cells were treated with 20.2 g of a 37.7% MeOH soln. at 21.4°C for 103 min. The H₂O₂ decompn. rate const. for permeabilized yeasts was 9 times higher than for the intacted ones.
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