Determination of in vitro angiotensin I converting enzyme inhibition activity of Chiropsalmus quadrigatus haeckel (Box Jellyfish) venom hydrolysate

Pamela Berilyn So, Mary Jho-Anne Corpuz and Oliver B Villaflores

Determination of in vitro angiotensin I converting enzyme inhibition activity of Chiropsalmus quadrigatus haeckel (Box Jellyfish) venom hydrolysate

Pamela Berilyn So, Mary Jho-Anne Corpuz and Oliver B Villaflores

University of Santo Tomas, Philippines

: J Pharm Drug Deliv Res

Abstract

Hypertension is a common problem among people worldwide and statistics from the World Health Organization shows that 21,000 Filipinos died of hypertensive heart disease in 2012 accounting for 3.7% of deaths that year. This number continues to rise showing an increasing trend from the year 2000 up to the present (WHO, 2015). ACE (angiotensin I converting enzyme) inhibitors are among the first-line treatment for high blood pressure. According to Balti (2015), peptides with ACE inhibitory activity have been found in several seafood hydrolysates and these peptides have shown potent antihypertensive activity. As these jellyfish are considered as waste, and even pests by most fishermen, utilizing it as a source of ACE inhibitory peptides is a better option instead of just throwing it away. The venom extract was obtained by centrifugation (Gyrozen Microcentrifuge 1730R, Korea) at 5000 rpm for 15 minutes at 4Ã‚Â°C, followed by ultrasonication (Power Sonic410, Korea), and it was further clarified by centrifugation at 20,000 x g for 1 hour at 4Ã‚Â°C. Protein content of 12.99 ÃŽÂ¼g/mL was determined using Bradford assay with Coomassie Brilliant BlueG-250 as indicator. The jellyfish venom extract was subjected to a two-step enzymatic hydrolysis with pepsin, followed by digestion with papain. The enzymatic hydrolysate was then subjected to column chromatography with Sephadex G-25 as the stationary phase and a linear gradient of sodium chloride was used for elution. The fractions were pooled together based on the Bradford assay. In vitro ACE inhibition was determined using ACE kit-WST from Dojindo Laboratories, and the absorbances were measured in triplicate using SH-1000 Lab Microplate reader (Corona Electric Co., Inc., Japan). Fractions 3, 4, and 7, exhibited high percent ACE inhibition of 86.49%, 85.71%, and 83.12%, respectively, which can be further purified to obtain potent ACE inhibitory peptides.