TY - JOUR
T1 - High enrichment of MMP-9 and carboxypeptidase a by tweezing adsorptive bubble separation (TABS)
AU - Haller, Dirk
AU - Ekici, Perihan
AU - Friess, Albrecht
AU - Parlar, Harun
N1 - Funding Information:
Acknowledgement Financial support by the German Federation of Industrial Research Associations “Otto von Guericke” (AiF) through the Research Association of the German Food Industry (FEI) is gratefully acknowledged (project 121 ZN).
PY - 2010/11
Y1 - 2010/11
N2 - 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.
AB - 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.
KW - Carboxypeptidase A
KW - Carboxypeptidases
KW - MMP-9
KW - Matrix metalloproteinases
KW - Tweezing adsorptive bubble separation
UR - http://www.scopus.com/inward/record.url?scp=78649669292&partnerID=8YFLogxK
U2 - 10.1007/s12010-010-8936-x
DO - 10.1007/s12010-010-8936-x
M3 - Article
C2 - 20229282
AN - SCOPUS:78649669292
SN - 0273-2289
VL - 162
SP - 1547
EP - 1557
JO - Applied Biochemistry and Biotechnology
JF - Applied Biochemistry and Biotechnology
IS - 6
ER -