High enrichment of MMP-9 and carboxypeptidase A by tweezing adsorptive bubble separation (TABS).

Dirk Haller, Perihan Ekici, Albrecht Friess, Harun Parlar

Chair for Biofunctionality, ZIEL-Research Center for Nutrition and Food Science, Technische Universität München, Gregor-Mendel-Strasse 2, Freising-Weihenstephan, Germany.

Applied biochemistry and biotechnology 2010 Nov

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Tweezing adsorptive bubble separation (TABS) was used as a method for the enrichment of matrix metalloproteinases (92-kDa type IV, gelatinase B (MMP-9)) and carboxypeptidase A (CPA) from dilute aqueous solutions. The method is based on the chelation of metalloenzymes applying 2-(carbamoylmethyl-(carboxymethyl)amino)acetic acid (ADA) coupled with an octyl part to form a surface active unit. MMP-9 could be enriched with an enrichment ratio of 12.0 and a recovery of 87.3%, and CPA could be enriched 18.8-fold and with 95.3% recovery. Both enzymes were enriched without significant losses of enzymatic activity. To verify that the enzymes were tweezed by ADA-C8 without abstraction of the zinc ions from the active center, TABS trials were additionally conducted with zinc ions in complex with ADA-C8, which revealed only negligible enrichment ratios of the enzymes (2.2 for MMP-9 and 0.2 for CPA). The results obtained impressively demonstrate that zinc-containing proteases can be enriched selectively and efficiently by TABS.

Citation

Dirk Haller, Perihan Ekici, Albrecht Friess, Harun Parlar. High enrichment of MMP-9 and carboxypeptidase A by tweezing adsorptive bubble separation (TABS). Applied biochemistry and biotechnology. 2010 Nov;162(6):1547-57