D-Penisilamin, D-Penisilamin disülfit ve N-Asetil-D-penisilamin’in Laktoperoksidaz Enzim Aktivitesi Üzerine İnhibisyon Etkileri

Abstract

Meme, tükürük ve diğer mukoza bezlerinden salgılanan laktoperoksidaz (LPO, E.C.1.11.1.7); antibakteriyel özelliğe sahip bir peroksidaz enzimidir. D-Penisilamin; Wilson hastalığı gibi birçok hastalığın tedavisinde kullanılan tiyol yan zincirine sahip bir amino asittir. Bu çalışmanın amacı, sığır laktoperoksidaz enzimine karşı D-Penisilamin, D-Penisilamin disülfit ve N-Asetil-D-penisilamin'in in vitro inhibisyon profillerini belirlemektir. LPO enzimi, % 62,25 verimle 357,92 kat afinite kromatografisi (Sepharose 4B-L-tirozin-sülfanamid) kullanılarak sığır sütünden saflaştırıldı. LPO; D-Penisilamin, D-Penisilamin disülfit ve N-Asetil-D-penisilamin tarafından etkili bir şekilde inhibe edildi. Bu moleküllerin IC50 değerleri sırasıyla 0,584, 0,207 ve 0,552 μM olarak bulundu. D-Penisilamin ve D-Penisilamin disülfit için yarışmalı inhibisyon gösterirken, N-Asetil-D-penisilamin için yarışmasız inhibisyon gösterdi.

Keywords

• Laktoperoksidaz,penisilamin,saflaştırma,inhibisyon

The Inhibition Effects of D-Penicillamine, D-Penicillamine disulfide and N-Acetyl-D-Penicillamine on Lactoperoxidase Enzyme Activity

Abstract

The lactoperoxidase (LPO, E.C.1.11.1.7) secreted from the breast, saliva and other mucous glands is a peroxidase enzyme with antibacterial properties. D-penicillamine is an amino acid with a thiol side chain, used in the treatment of many diseases such as Wilson disease. The aim of this study was to determine in vitro inhibition profiles of D-Penicillamine, D-Penicillamine disulfide and N-Acetyl-D-penicillamine against bovine lactoperoxidase enzyme. LPO enzyme was isolated from bovine milk using affinity chromatography (Sepharose 4B-L-tyrosine-sulphanamide) 357.92 fold with a yield of 62.25%. LPO was effectively inhibited by D-Penicillamine, N-Acetyl-D-penicillamine and D-Penicillamine disülfit. IC50 values of these molecules were found as 0.584, 0.207 and 0.552 𝜇M, respectively. D-Penicillamine, and D-Penicillamine disülfit exhibited competitive inhibition, and N-Acetyl-D-penicillamine showed noncompetitive inhibition.

Keywords

• Laktoperoksidaz,penisilamin,saflaştırma,inhibisyon

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