Artículos de revistas
Purification Of Microbial β-galactosidase From Kluyveromyces Fragilis By Bioaffinity Partitioning
Registro en:
Revista De Microbiologia. , v. 30, n. 4, p. 324 - 331, 1999.
13714
2-s2.0-22844448692
Autor
Da Silva M.E.
Franco T.T.
Institución
Resumen
This work investigated the partitioning of β-galactosidase from Kluyveromyces fragilis in aqueous two-phase systems (ATPS) by bioaffinity. PEG 4000 was chemically activated with thresyl chloride, and the biospecific ligand p-aminophenyl 1-thio-β-D-galactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme β-galactosidase from Kluyveromyces fragilis was developed. In the first step, a system composed of 6% PEG 4000-APGP and 8% dextran 505 was used, where β-galactosidase was strongly partitioned to the top phase (K = 2,330). In the second step, a system formed of 13% PEG-APGP and 9% phosphate salt was used to revert the value of the partition coefficient of β-galactosidase (K = 2 x 10-5) in order to provide the purification and recovery of 39% of the enzyme in the bottom salt-rich phase. 30 4 324 331 Aguiñaga-Díaz, P.A., Guzmán, R.Z., Affinity partitioning of metal ions in aqueous polyethylene glycol/salt two-phase systems with PEG-modified dictators (1996) Sep. Sci. Technol., 31 (10), pp. 1483-1499 Albertsson, P.Å., (1971) Partition of Cell Particles and Macromolecules. 2nd Ed., p. 323. , New York: Interscience Albertsson, P.Å., Tjemeld, F., Phase Diagrams (1994) Meth. Enzumol., 228, pp. 3-13 Baskir, J.N., Hatton, T.A., Suter, U.W., Protein partitioning in two-phase aqueous polymer systems (1989) Biotechol. Bioeng., 34 (4), pp. 541-558 Birkenmeier, G., Partitioning of blood proteins using immobilized dyes (1994) Meth. 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