RESUMEN

The behavior of xylanase extraction in a packed column using polyethylene glycol (PEG) 4000 and dipotassium phosphate was studied. The possibility of using the packed column in continuous operations for enzyme extraction was studied since the previous work had only addressed the semi-continuous extraction of enzyme. The influence of several kinds of packings, Raschig rings, glass spheres and polystyrene rings were studied as well the superficial velocity ratio of the salt and the PEG phases. Packed column showed a good efficiency of overall mass transfer coefficient, around three times higher than sieve plate column, for xylanase extraction. The best selectivity was obtained with the polystyrene ring where 94% of xylanase was recovery to the polymeric whereas just 3% of contaminant was recovery to this phase. The residence time distribution was adjusted by the Model of Reactors in Series.

Asunto(s)

Cromatografía Liquida/instrumentación , Xilosidasas/metabolismo

RESUMEN

Aqueous two-phase systems (ATPSs) have long been used for biomolecule partitioning; these systems offer the possibility of using continuous or semicontinuous extraction processes. They require relatively simple equipment like spray or sieve plate columns that can be adapted for use in ATPSs. The aim of this work was to study the semicontinuous extraction of a model enzyme, xylanase, in spray and sieve plate columns, since, unlike centrifugal contactors, the cost of construction and maintenance of this equipment is low and it is easy to operate. For the spray column, the dispersed phase hold-up and overall mass transfer coefficients K(D) a were evaluated for different column heights and for different superficial velocities of the dispersed phase (light phase). Results indicated that an increase in superficial velocity in the range of 0-0.18 mm/s of the dispersed phase had a positive effect on K(D) a and on hold-up in all column heights studied, 75, 161 and 246 mm. For the same superficial velocity of the dispersed phase, the larger the hold-up was, the shorter the column. For the sieve plate column, the effects of the superficial velocity of the dispersed phase and the number of plates were also studied. Results showed that the K(D) a and hold-up increased with an increase in both parameters. The selectivity of separation of xylanase and BSA (model contaminant) was very high, since 60% of the enzyme was extracted in the light phase, whereas no significant amount of BSA was extracted. The possibility of using the sieve plate column in continuous operation for enzyme extraction was studied because previous work had only addressed the semicontinuous extraction of enzyme. The residence time distribution of the PEG phase using different superficial velocities of the salt phase was studied in continuous operation. The time required to reach the steady state was 40 min, and 70% of the xylanase was recovered. It was found that the Modified Power Spline software was well adjusted to the experimental results.

Asunto(s)

Cromatografía Liquida/instrumentación , Cromatografía Liquida/métodos , Programas Informáticos , Xilosidasas/aislamiento & purificación , Animales , Bovinos , Concentración de Iones de Hidrógeno , Polietilenglicoles/química , Albúmina Sérica/química , Agua/química , Xilosidasas/química

RESUMEN

Chromatographic quantitative analyses of aqueous samples containing dilute concentrations of organic solutes are frequently confounded by sorption phenomena, particularly for vapor samples and aged syringes. A novel apparatus was developed for overcoming this problem; it utilizes continual circulation of gas through a thermostated syringe for sampling, followed by a needleless injection into the chromatograph. Precision and reproducibility well under 0.5% can be obtained in this way. This method was applied to a determination of the solubility of isopentyl acetate in water (2600 ppm) by following the equilibrium vapor response for increasing liquid concentrations. It was also used to determine equilibrium partition coefficients for C1-C5 n-acetates between air and water, over a range of temperatures from 25 to 40 degrees C.