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

Marine Drugs

Isabel Rodríguez Amado¹  José Antonio Vázquez¹  Pilar González¹  Diego Esteban-Fernández²  Mónica Carrera³ 

[1] Group of Recycling and Valorisation of Waste Materials (REVAL), Marine Research Institute (IIM-CSIC), r/Eduardo Cabello, 6. Vigo, Galicia 36208, Spain; E-Mails:;Department of Chemistry, Humboldt-Universitaet zu Berlin, Brook-Taylor Strasse 2, Berlin 12489, Germany; E-Mail:

关键词: ultrafiltration;    proteolysis;    ACE inhibitory peptides;    cuttlefish byproducts;    peptide identification;    HPLC-ESI-MS;   

DOI  :  10.3390/md12031390

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.