Revista de Microbiologia | |
Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning | |
Universidade Estadual de Campinas, unicamp, Campinas, Brasil1  Universidade Estadual de Campinas, Campinas, Brasil1  Silva, Maria Estela da1  Franco, Telma Teixeira1  | |
关键词: : b-galactosidase; aqueous two-phase systems; protein purification; downstream-processing; affinity; | |
DOI : 10.1590/S0001-37141999000400006 | |
学科分类:农业科学(综合) | |
来源: Sociedade Brasileira de Microbiologia / Brazilian Society for Microbiology | |
【 摘 要 】
This work investigated the partitioning of b-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-b-D-galactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme b-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 b-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 b-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.
【 授权许可】
Unknown
【 预 览 】
Files | Size | Format | View |
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RO201912010132213ZK.pdf | 433KB | download |