alexa One step partial purification of β-D-galactosidase from Kluyveromyces marxianus CDB 002 using STREAMLINE-DEAE


Journal of Food & Industrial Microbiology

Author(s): Mara Jos Artolozaga, Rainer Jonas, Andra L Schneider, Sandra A Furlan, Maria de Ftima CarvalhoJona

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The intracellular enzyme β-D-galactosidase provides interesting applications in the dairy industry, which are able to solve problems related to product processing, or can alleviate lactose intolerance in some populations. In order to obtain a technical enzyme, yeast cells of Kluyveromyces marxianus CDB 002 were disrupted by high pressure homogenization and an innovative chromatographic technique was tested for the recovery of β-D-galactosidase. A STREAMLINE 25 column, containing 65 ml STREAMLINE-DEAE was equilibrated with 50 mM potassium phosphate buffer pH 7.5 at an upward flow of 250 cmh–1. 100–200 ml cell homogenate were applied onto the expanded gel. After unbound proteins and cellular debris were washed out, the bed was allowed to sediment and β-D-galactosidase was eluted with a downward flow of 0.2 M NaCl in the same buffer. A 6-fold purification factor was achieved with 63% activity recovery, while removing cell debris at a single step, thus avoiding a centrifugation step. Concentration and volume of the applied sample affected purification and gel performance. The results presented show STREAMLINE-DEAE chromatography to be an interesting method for the production of β-D-galactosidase as a technical enzyme, since it can also be applied on a large scale without much modification.

This article was published in Bioseparation and referenced in Journal of Food & Industrial Microbiology

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