Lactoperoxidase Enzyme Purified from Different Milk Sources: Inhibition Profile of Capsaicin and Pyrogallol

Abstract

Peroxidases (POD) have an important use in metabolic functions, enzymatic reactions and clinical

diagnoses, especially in the food and pharmaceutical industry. Mammalian POD enzymes, Lactoperoxidase (LPO)

(hydrogen peroxide oxidoreductase E.C. 1.11.1.7) is localized in milk, saliva and tears, while myeloperoxidase is

localized in leukocytes and platelets. LPO enzyme catalyses the conversion of hypothiocyanate with antibacterial

properties of thiocyanate in the presence of hydrogen peroxide. The purpose of this study was to determine the in

vitro inhibition effects of capsaicin and pyrogallol on LPO enzyme purified from different milk sources (Bovine,

buffalo, sheep and goat). To determine the inhibition effects of capsaicin and pyrogallol on LPO, LPO enzyme

was purified from different mammalian milk and then, Lineweaver-Burk graphs were drawn for each inhibitor

by measuring enzyme activities; Ki values and inhibition types were determined from these plotted graphs. The

Ki values of capsaicin and pyrogallol were found in ranging of 0.0035-36.178 µM. Pyrogallol was shown the

most effective inhibitor feature with a non-competitive inhibition type with 0.0035 ± 0.0012 μM Ki value on LPO

enzyme purified from sheep milk.

Keywords

• Capsaicin,enzyme inhibition,lactoperoxidase,mammalian milk,pyrogallol

Laktoperoksidaz Enziminin Farklı Kaynaklardan Saflaştırılması: Kapsaisin ve Pirogallol’ün İnhibisyon Profili

Abstract

Peroksidazlar (POD), gıda ve ilaç endüstrisi başta olmak üzere metabolik fonksiyonlar, enzimatik

reaksiyonlar ve klinik teşhislerde önemli kullanım alanına sahiptirler. Memeli POD enzimlerinden laktoperoksidaz

(LPO) süt, tükrük ve gözyaşında (hidrojen peroksit oksidoredüktaz E.C 1.11.1.7) lokalize olurken, miyeloperoksidaz

lökositler ve trombositlerde lokalizedir. LPO enzimi hidrojen peroksit eşliğinde tiyosiyanatın antibakteriyel

özelliklere sahip hipotiyosiyanata dönüşümünü katalizler. Bu çalışmanın amacı, farklı memeli (Sığır, manda, koyun

ve keçi) sütlerinden saflaştırılmış LPO enzimi üzerine kapsaisin ve pirogallolün in vitro etkilerini belirlemektir.

Kapsaisin ve pirogallolün LPO enzimi üzerindeki inhibisyon etkisini belirlemek için herbir memeli sütünden

LPO enzimi saflaştırıldı, daha sonra, enzim aktiviteleri ölçülerek herbir inhibitör için Lineweaver-Burk grafikleri

çizildi; Ki sabiti ve inhibisyon tipleri bu çizilen grafiklerden hesaplandı. Kapsaisin ve pirogallolün Ki değerleri

0.0035-36.178 μM aralığında bulundu. Pirogallol, en etkili inhibitör özelliğini yarışmasız inhibisyon tipi ile koyun

sütünden saflaştırılmış LPO enzimi üzerine 0.0035 ± 0.0012 μM Ki değeri ile göstermiştir.

Keywords

• Enzim inhibisyonu,kapsaisin,laktoperoksidaz,memeli sütleri,pirogallol

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