Extraction of Intracellular Proteins from Kluyveromyces lactis
Manuel Becerra, Esther Rodríguez-Belmonte, Ma Esperanza Cerdán and Ma Isabel González Siso*
Departamento de Bioloxía Celular e Molecular, Facultade de Ciencias Campus da Zapateira s/n. E-15071- La Coruña, Spain
Article history:
Received November 2, 1999
Accepted March 23, 2001
Key words:
β-galactosidase, purification, extraction, sonication, Kluyveromyces lactis
Summary:
Several preparation methods of crude extracts of yeasts were compared in order to obtain a good recovery of intracellular proteins and a sample quality suitable for further chromatographic analysis as exemplified by β-galactosidase or lactase. Sonication was found to be the most effective method and the physiological state of the cells and growth conditions at the time of extraction are essential parameters for obtaining good yield. The procedure described is simple, rapid and applicable to other enzymes of interest.
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