Investigation of Anti-enzyme Activities of Primrose (Oenothera biennis) and Sweet Almond (Prunus dulcis Mill.) Oils

Abstract

In our study, the question of what might be the effects of Primrose (Oenothera biennis) and sweet almond (Prunus dulcis Mill.) oils on two different enzyme activities was asked. The first enzyme in our study is carbonic anhydrase I-II isoenzymes that play a very important role in many physiological events, by transforming catalysis of CO2 hydration and HCO3-dehydration in living things, by creating an intracellular bicarbonate buffer system. The second enzyme, the acetylcholinesterase enzyme, which was located in the synapse between the muscle cell and nerve, catalyzes the decomposition of the acetylcholine molecule. Inhibitors of both enzymes have the potential to be drugs. Therefore, the inhibition effect of the specified oils was investigated. For this purpose, the first carbonic anhydrase I-II isoenzyme was purified by the sepharose-4B-L tyrosine-sulfonamide affinity column. Then, by looking at enzyme activity in at least five different inhibitor concentrations, effects of evening primrose (Oenothera biennis) and sweet almond (Prunus dulcis Mill.) oils on enzyme activities were investigated. Finally, IC50 values of Primrose (Oenothera biennis) and sweet almond (Prunus dulcis Mill.) oils were determined by drawing activity%-[I] graph. Carbonic anhydrase I isoenzyme was purified from human erythrocytes 119 times in 20.12% yield, while carbonic anhydrase II isoenzyme was purified 535.72 times in 83.05% yield. IC50 values of the primrose oils (Oenothera biennis) on carbonic anhydrase I-II and acetylcholinesterase enzyme activity were 0,1950, 0,1406 and 0,1097 mg / mL, respectively. IC50 values of the sweet almond oil (Prunus dulcis Mill.) on carbonic anhydrase I-II and acetylcholine esterase enzyme activity were 0.0345, 0.0266 and 0.0394 mg/mL respectively. When both oils used in the study are compared, we see that sweet almond oil (Prunus dulcis Mill.) is more effective on both enzyme activities. Therefore, instead of synthetic drugs, sweet almond (Prunus dulcis Mill.) oil is thought may be used as a supplement in the treatment of diseases caused by the enzymes in question.

Keywords

• Acetylcholinesterase,carbonic anhydrase,in vitro

Çuha Çiçeği (Oenothera biennis) ve Tatlı Badem (Prunus dulcis Mill) Yağlarının Anti-enzim Aktivitelerinin Araştırılması

Öz

Çalışmamızda iki farklı enzim aktivitesi üzerine Çuha çiçeği (Oenothera biennis) ile tatlı badem (Prunus dulcis Mill.) yağlarının etkileri ne olabilecek sorusu cevaplanmak istendi. Kullandığımız ilk enzim canlılarda CO2’in hidratasyonu ve HCO3-‘ın dehidratasyonunu dönüşümlü olarak katalizleyerek, hücre içi bikarbonat tampon sistemini oluşturarak, birçok fizyolojik olayda oldukça önemli rol alan karbonik anhidraz I-II izoenzimleridir. İkinci enzim ise asetilkolin molekülünün ayrışmasını katalizleyen, kas hücresi ve sinir arasındaki sinapsta yer alan asetilkolinesteraz enzimidir. Her iki enzimin inhibitörleri, ilaç olma potansiyeli taşımaktadır. Bu nedenle belirtilen yağların inhibisyon etkisi araştırıldı. Bu amaçla ilk olarak karbonik anhidraz I-II izoenzimi sefaroz-4B-L-tirosin-sülfanilamid afinite kolonu ile saflaştırıldı. Ardından en az beş farklı inhibitör konsantrasyonunda enzim aktivitesi bakılarak Çuha çiçeği (Oenothera biennis) ve tatlı badem (Prunus dulcis Mill.) yağlarının enzim aktiviteleri üzerindeki etkileri araştırıldı. Son olarak %aktivite-[I] grafiği çizilerek Çuha çiçeği (Oenothera biennis) ve tatlı badem (Prunus dulcis Mill.) yağlarının IC50 değerleri tespit edildi. İnsan eritrositlerinden karbonik anhidraz I izoenzimi %20,12 verimle 119 kat saflaştırılırken, karbonik anhidraz II izoenzimi %83,05 verimle 535,72 kat saflaştırıldı. Çuha çiçeği (Oenothera biennis) yağının karbonik anhidraz I-II ve asetilkolinesteraz enzim aktivitesi üzerindeki IC50 değerleri sırasıyla 0,1950, 0,1406 ve 0,1097 mg/mL olarak, tatlı badem (Prunus dulcis Mill.) yağının karbonik anhidraz I-II ve asetilkolin esteraz enzim aktivitesi üzerindeki IC50 değerleri ise sırasıyla 0,0345, 0,0266 ve 0,0394 mg/mL olarak tespit edildi. Çalışmada kullanılan her iki yağ karşılaştırıldığında, tatlı badem (Prunus dulcis Mill.) yağının her iki enzim aktivitesi üzerinde daha etkili olduğunu görülmektedir. Bu nedenle sentetik ilaçlar yerine tatlı badem (Prunus dulcis Mill.) yağının söz konusu enzimlerin sebep olduğu hastalıkların tedavisinde takviye ilaç olarak kullanılabileceği düşünülmektedir.

Anahtar Kelimeler

• Asetilkolinesteraz,karbonik anhidraz,in vitro

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