Isolation of angiotensin I converting enzyme inhibitory ... of...Isolation of angiotensin I...
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Isolation of angiotensin I converting enzyme inhibitory peptides from a fish by-products hydrolysate Tensideal ® Véronique Lahogue 1,2 , Karine Vallée-Rehel 1 , Mylène Cuart 1 , Dominique Haras 1 and Patrick Allaume 2 1 Laboratoire de Biotechnologie et Chimie Marines – Université de Bretagne Sud – BP 92116 – 56321 Lorient - France; 2 IDMER – 2 rue Batelière – 56100 Lorient - France Introduction Many ACE inhibitory peptides have been isolated from various protein hydrolysates such as casein, soybean or fish protein. The purpose of this work is to isolate ACE inhibitory peptides derived from an enzymatic hydrolysate of fish by-products named Tensideal®, which is actually produced industrially by IDMER. The measurement of ACE inhibitory activity was performed using a HPLC method consisting on measuring the levels of the substrate (FAPGG (N-(3-[2- furylacryloyl)-Phe-Gly-Gly)) or the product formed (FAP) with and without inhibitor. By this method, the hydrolysate Tensideal® showed an IC50 of 65μg of peptides while the reference Captopril® showed an IC50 of 0.3ng. To isolate ACE inhibitory peptides from the hydrolysate, different chromatographic methods were successively applied to select step by step the most active fraction. Gel filtration has demonstrated that major peptides have molecular weight less than 1000 Da and some tests realised by C18 HPLC have showed that active peptides were very polar molecules. After gel filtration, SPE and LC-MS, we succeeded in identifying an active fraction with an interesting mass profile. The interpretation of the mass spectrum is actually in progress. Measurement of ACE inhibitory activity The compound FAP resulting of the FAPGG hydrolysis by ACE is quantified with and without inhibitor and results are compared to determine the ACE inhibition degree. Fig. 1. HPLC Measurement of ACE inhibitory activity : % ACE inhibition = 54.6%. FAPGG : RT = 7.880 min - %area = 13.02% FAP : RT = 14.821 min - %area = 86.98% FAPGG : RT = 7.899 min - %area = 60.53% FAP : RT = 14.787 min - %area = 39.47% CONTROL : WITHOUT INHIBITOR ASSAY : WITH INHIBITOR S. Del Valle et al. (eds.), Peptides for Youth: The Proceedings of the 20th American Peptide Symposium, 483 DOI: 10.1007/978-0-387-73657-0_208, © Springer Science+Business Media, LLC 2009
Transcript of Isolation of angiotensin I converting enzyme inhibitory ... of...Isolation of angiotensin I...
Isolation of angiotensin I converting enzyme inhibitory peptides from a fish by-products hydrolysate Tensideal®
Véronique Lahogue1,2, Karine Vallée-Rehel1, Mylène Cuart1, Dominique Haras1 and Patrick Allaume2
1Laboratoire de Biotechnologie et Chimie Marines – Université de Bretagne Sud – BP 92116 – 56321 Lorient - France; 2IDMER – 2 rue Batelière – 56100 Lorient - France
Introduction Many ACE inhibitory peptides have been isolated from various protein hydrolysates such as casein, soybean or fish protein. The purpose of this work is to isolate ACE inhibitory peptides derived from an enzymatic hydrolysate of fish by-products named Tensideal®, which is actually produced industrially by IDMER.
The measurement of ACE inhibitory activity was performed using a HPLC method consisting on measuring the levels of the substrate (FAPGG (N-(3-[2-furylacryloyl)-Phe-Gly-Gly)) or the product formed (FAP) with and without inhibitor. By this method, the hydrolysate Tensideal® showed an IC50 of 65µg of peptides while the reference Captopril® showed an IC50 of 0.3ng.
To isolate ACE inhibitory peptides from the hydrolysate, different chromatographic methods were successively applied to select step by step the most active fraction. Gel filtration has demonstrated that major peptides have molecular weight less than 1000 Da and some tests realised by C18 HPLC have showed that active peptides were very polar molecules. After gel filtration, SPE and LC-MS, we succeeded in identifying an active fraction with an interesting mass profile. The interpretation of the mass spectrum is actually in progress. Measurement of ACE inhibitory activity The compound FAP resulting of the FAPGG hydrolysis by ACE is quantified with and without inhibitor and results are compared to determine the ACE inhibition degree. Fig. 1. HPLC Measurement of ACE inhibitory activity : % ACE inhibition = 54.6%.
FAPGG : RT = 7.880 min - %area = 13.02%FAP : RT = 14.821 min - %area = 86.98%
FAPGG : RT = 7.899 min - %area = 60.53% FAP : RT = 14.787 min - %area = 39.47%
CONTROL : WITHOUT INHIBITOR ASSAY : WITH INHIBITOR
S. Del Valle et al. (eds.), Peptides for Youth: The Proceedings of the 20th American Peptide Symposium, 483DOI: 10.1007/978-0-387-73657-0_208, © Springer Science+Business Media, LLC 2009
