Identification of the Major ACE-Inhibitory Peptides Produced by Enzymatic Hydrolysis of a Protein Concentrate from Cuttlefish Wastewater

Isabel Amado,Carmen Piñeiro,José Vázquez,Pilar González,Mónica Carrera,Diego Esteban-Fernández

doi:10.3390/md12031390

Isabel Amado, Carmen Piñeiro + Show 4 more

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https://doi.org/10.3390/md12031390

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Journal: Marine Drugs	Publication Date: Mar 10, 2014
Citations: 65	License type: CC BY 3.0

Affiliation: ETH Zurich, Humboldt-Universität zu Berlin

Abstract

The aim of this work was the purification and identification of the major angiotensin converting enzyme (ACE) inhibitory peptides produced by enzymatic hydrolysis of a protein concentrate recovered from a cuttlefish industrial manufacturing effluent. This process consisted on the ultrafiltration of cuttlefish softening wastewater, with a 10 kDa cut-off membrane, followed by the hydrolysis with alcalase of the retained fraction. Alcalase produced ACE inhibitors reaching the highest activity (IC50 = 76.8 ± 15.2 μg mL−1) after 8 h of proteolysis. Sequential ultrafiltration of the 8 h hydrolysate with molecular weight cut-off (MWCO) membranes of 10 and 1 kDa resulted in the increased activity of each permeate, with a final IC50 value of 58.4 ± 4.6 μg mL−1. Permeate containing peptides lower than 1 kDa was separated by reversed-phase high performance liquid chromatography (RP-HPLC). Four fractions (A–D) with potent ACE inhibitory activity were isolated and their main peptides identified using high performance liquid chromatography coupled to an electrospray ion trap Fourier transform ion cyclotron resonance-mass spectrometer (HPLC-ESI-IT-FTICR) followed by comparison with databases and de novo sequencing. The amino acid sequences of the identified peptides contained at least one hydrophobic and/or a proline together with positively charged residues in at least one of the three C-terminal positions. The IC50 values of the fractions ranged from 1.92 to 8.83 μg mL−1, however this study fails to identify which of these peptides are ultimately responsible for the potent antihypertensive activity of these fractions.

Highlights

Cardiovascular diseases (CVD) are the main cause of death globally, accounting for approximately17 million deaths a year, nearly one third of the total [1]
According to the World Health Organization, most cardiovascular diseases can be prevented by addressing risk factors, such as tobacco use, unhealthy diet and obesity, physical inactivity, high blood pressure, diabetes, and raised lipids [2]
Blood pressure is regulated by the renin-angiotensin system, wherein, the angiotensin converting enzyme (ACE) hydrolyses biologically inactive angiotensin I to the potent vasoconstrictor angiotensin

Summary

Introduction

Cardiovascular diseases (CVD) are the main cause of death globally, accounting for approximately. 17 million deaths a year, nearly one third of the total [1]. According to the World Health Organization, most cardiovascular diseases can be prevented by addressing risk factors, such as tobacco use, unhealthy diet and obesity, physical inactivity, high blood pressure, diabetes, and raised lipids [2]. Among all CVD, complications of hypertension account for 9.4 million deaths worldwide every year [3] and are the most common risk factors of heart diseases. Hypertension is considered a major health problem worldwide. Blood pressure is regulated by the renin-angiotensin system, wherein, the angiotensin converting enzyme (ACE) hydrolyses biologically inactive angiotensin I to the potent vasoconstrictor angiotensin. The renin-angiotensin system has become a key target for drugs combating hypertension [4] and, various synthetic

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Conclusion