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

Appl Biochem Biotechnol. 2010 Nov;162(6):1547-57. doi: 10.1007/s12010-010-8936-x. Epub 2010 Mar 14.

Abstract

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carboxypeptidases A / analysis
  • Carboxypeptidases A / isolation & purification*
  • Chemistry Techniques, Analytical / methods*
  • Matrix Metalloproteinase 9 / analysis
  • Matrix Metalloproteinase 9 / isolation & purification*

Substances

  • Carboxypeptidases A
  • Matrix Metalloproteinase 9