A new approach for affinity-based purification of horseradish peroxidase

Almaz Z., ÖZTEKİN A., Abul N., Gerni S., Erel D., Kocak S. M., ...Daha Fazla

BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, cilt.68, sa.1, ss.102-113, 2021 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 68 Sayı: 1
  • Basım Tarihi: 2021
  • Doi Numarası: 10.1002/bab.1899
  • Dergi Adı: BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Agricultural & Environmental Science Database, Applied Science & Technology Source, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Compendex, Computer & Applied Sciences, EMBASE, Environment Index, Food Science & Technology Abstracts, INSPEC, MEDLINE, Veterinary Science Database
  • Sayfa Sayıları: ss.102-113
  • Anahtar Kelimeler: affinity purification, benzohydrazide, horseradish peroxidase, GLUTATHIONE-S-TRANSFERASE, TURNIP PEROXIDASE, LACTOPEROXIDASE, ENZYMES, BUTYRYLCHOLINESTERASE, ACETYLCHOLINESTERASE, MEMBRANE, PROTEINS, L.
  • Atatürk Üniversitesi Adresli: Evet

Özet

We have developed efficient procedure for isolation of horseradish peroxidase (HRP) using aminobenzohydrazide-based affinity chromatography. Sepharose 4B-bounded aminobenzohydrazides are suitable for long-term use and large-scale purification. In this study, 26 aminobenzohydrazide derivatives were synthesized, characterized and defined as new HRP inhibitors. In addition, detailed inhibition effects of these molecules on HRP enzyme were investigated. Affinity matrix was formed by bonding aminobenzohydrazides, which exhibited inhibitory activity to sepharose-4B-l-tyrosine. HRP was isolated from crude homogenate in single step and purification factors were recorded as 1,151-fold (recovery of 8.5%) with 4-amino 3-bromo benzohydrazide and as 166.16-fold (recovery of 16.67 %) with 3-amino 4-chloro benzohydrazide.