Journalartikel

Autorenliste: Gerken, BM; Wattenbach, C; Linke, D; Zorn, H; Berger, RG; Parlar, H

Jahr der Veröffentlichung: 2005

Seiten: 6113-6117

Zeitschrift: Analytical Chemistry

Bandnummer: 77

Heftnummer: 19

ISSN: 0003-2700

DOI Link: https://doi.org/10.1021/ac050977s

Verlag: American Chemical Society

Abstract: 
A novelly developed tweezing-adsorptive bubble separation (ABS) method for the enrichment of metalloenzymes (laccase C and horseradish peroxidase) is introduced. The method is based on the chelation of the enzymes' active center and can also be applied for analysis. N-(2-Acetamido)iminodiacetic acid served as a chelator and was synthesized with an octyl unit to become ADA-C8. Laccase was enriched 13.3-fold (66.31% recovery) and HPOX 17.8-fold (85.34%) without a significant loss of enzymatic activity. To prove that the entire enzyme is tweezed at the active center, ABS trials were done using ADA-C8 already complexed with Cu2+ and Fe3+. As only marginal enrichment occurred (ER laccase, 0.17; ER HPOX, 0.44), no chelating effect was concluded. It was determined how the chelation toward the active center was directed by applying other chelators such as EDTA, NTA, NN-dimethylaminoglycine, oxalic acid, malonic acid, adipinic acid, and tripropylamine, which are similar in structure to ADA-C8. The results concluded that the chelation is 3-fold coordinated on the type 1 copper center of laccase, whereas that of HPOX only 1-fold at Fe3+ and additionally at the cationic amino acid arginine, which is also located at the active center. Tweezing-ABS has been proven to selectively and effectively enrich metalloenzymes.

Harvard-Zitierstil: Gerken, B., Wattenbach, C., Linke, D., Zorn, H., Berger, R. and Parlar, H. (2005) Tweezing-adsorptive bubble separation. Analytical method for the selective and high enrichment of metalloenzymes, Analytical Chemistry, 77(19), pp. 6113-6117. https://doi.org/10.1021/ac050977s

APA-Zitierstil: Gerken, B., Wattenbach, C., Linke, D., Zorn, H., Berger, R., & Parlar, H. (2005). Tweezing-adsorptive bubble separation. Analytical method for the selective and high enrichment of metalloenzymes. Analytical Chemistry. 77(19), 6113-6117. https://doi.org/10.1021/ac050977s