Angiotensin I-converting enzyme (ACE) inhibitory activities of sardinelle (Sardinella aurita) by-products protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases.

The angiotensin I-converting enzyme (ACE) inhibitory activities of protein hydrolysates prepared from heads and viscera of sardinelle (Sardinella aurita) by treatment with various proteases were investigated. Protein hydrolysates were obtained by treatment with Alcalase(®), chymotrypsin, crude enzyme preparations from Bacillus licheniformis NH1 and Aspergillus clavatus ES1, and crude enzyme extract from sardine (Sardina pilchardus) viscera. All hydrolysates exhibited inhibitory activity towards ACE. The alkaline protease extract from the viscera of sardine produced hydrolysate with the highest ACE inhibitory activity (63.2±1.5% at 2mg/ml). Further, the degrees of hydrolysis and the inhibitory activities of ACE increased with increasing proteolysis time. The protein hydrolysate generated with alkaline proteases from the viscera of sardine was then fractionated by size exclusion chromatography on a Sephadex G-25 into eight major fractions (P1-P8). Biological functions of all fractions were assayed, and P4 was found to display a high ACE inhibitory activity. The IC50 values for ACE inhibitory activities of sardinelle by-products protein hydrolysates and fraction P4 were 1.2±0.09 and 0.81±0.013mg/ml, respectively. Further, P4 showed resistance to in vitro digestion by gastrointestinal proteases. The amino acid analysis by GC/MS showed that P4 was rich in phenylalanine, arginine, glycine, leucine, methionine, histidine and tyrosine. The added-value of sardinelle by-products may be improved by enzymatic treatment with visceral serine proteases from sardine.

[1]  J. Mayne,et al.  Food and Agriculture Organisation , 1948, Nature.

[2]  R. Collins,et al.  Blood pressure, stroke, and coronary heart disease Part 2, short-term reductions in blood pressure: overview of randomised drug trials in their epidemiological context , 1990, The Lancet.

[3]  R. Collins,et al.  Blood pressure, stroke, and coronary heart disease Part 1, prolonged differences in blood pressure: prospective observational studies corrected for the regression dilution bias , 1990, The Lancet.

[4]  W. Horwitz.  Official Methods of Analysis , 1980 .

[5]  E. Seki,et al.  Inhibition of angiotensin I-converting enzyme by Bacillus licheniformis alkaline protease hydrolyzates derived from sardine muscle. , 1993, Bioscience, biotechnology, and biochemistry.

[6]  L. Skeggs,et al.  THE PREPARATION AND FUNCTION OF THE HYPERTENSIN-CONVERTING ENZYME , 1956, The Journal of experimental medicine.

[7]  H. Fujita,et al.  Classification and Antihypertensive Activity of Angiotensin I-Converting Enzyme Inhibitory Peptides Derived from Food Proteins , 2000 .

[8]  Young-Jun Choi,et al.  Isolation and characterization of a novel angiotensin I-converting enzyme inhibitory peptide derived from the edible mushroom Tricholoma giganteum , 2004, Peptides.

[9]  N. Yamamoto,et al.  Purification and characterization of angiotensin I-converting enzyme inhibitors from sour milk. , 1995, Journal of dairy science.

[10]  M. Yasuda,et al.  Production of angiotensin I-converting enzyme inhibitory peptides from soybean protein with Monascus purpureus acid proteinase , 2005 .

[11]  R. Fitzgerald,et al.  Identification of a novel angiotensin‐I‐converting enzyme inhibitory peptide corresponding to a tryptic fragment of bovine β‐lactoglobulin , 1997, FEBS letters.

[12]  Yong-hui Shi,et al.  Angiotensin I- converting enzyme inhibitory peptides derived from food proteins and their physiological and pharmacological effects , 2004 .

[13]  B. Rasco,et al.  Biochemical and functional properties of Atlantic salmon (Salmo salar) muscle proteins hydrolyzed with various alkaline proteases. , 2000, Journal of agricultural and food chemistry.

[14]  E. Li-Chan,et al.  Functional Properties of Fish Protein Hydrolysate from Herring (Clupea harengus) , 1999 .

[15]  F. Shahidi,et al.  Angiotensin I converting enzyme inhibitory peptides purified from bovine skin gelatin hydrolysate. , 2001, Journal of agricultural and food chemistry.

[16]  G. Quaglia,et al.  Enzymic solubilisation of proteins of sardine (sardina pilchardus) by commercial proteases , 1987 .

[17]  M. Nasri,et al.  Purification and characterization of trypsin from the viscera of sardine (Sardina pilchardus) , 2007 .

[18]  Fereidoon Shahidi,et al.  Antioxidative activity and functional properties of protein hydrolysate of yellow stripe trevally (Selaroides leptolepis) as influenced by the degree of hydrolysis and enzyme type , 2007 .

[19]  S. Maruyama,et al.  Angiotensin I-converting enzyme inhibitory activity and insulin secretion stimulative activity of fermented fish sauce. , 2003, Journal of bioscience and bioengineering.

[20]  D. Kwon,et al.  Purification and Identification of Angiotensin-I Converting Enzyme Inhibitory Peptide from Kidney Bean Protein Hydrolyzate , 1999 .

[21]  E. G. Erdös,et al.  Angiotensin I converting enzyme. , 1975, Circulation research.

[22]  J. Adler-Nissen,et al.  Enzymic Hydrolysis of Food Proteins , 1986 .

[23]  F. Netto,et al.  Effect of heat and enzymatic treatment on the antihypertensive activity of whey protein hydrolysates , 2007 .

[24]  Garland R. Marshall,et al.  Validated ligand mapping of ACE active site , 2005, J. Comput. Aided Mol. Des..

[25]  M. Linder,et al.  Influence of Hydrolysis Degree on the Functional Properties of Salmon Byproducts Hydrolysates , 2004 .

[26]  C. Berne,et al.  A comparison of the effects of hydrochlorothiazide and captopril on glucose and lipid metabolism in patients with hypertension. , 1989, The New England journal of medicine.

[27]  G. Huet,et al.  Sequential GC/MS analysis of sialic acids, monosaccharides, and amino acids of glycoproteins on a single sample as heptafluorobutyrate derivatives. , 2003, Biochemistry.

[28]  M. Nasri,et al.  Purification and characterization of an alkaline serine-protease produced by a new isolated Aspergillus clavatus ES1 , 2007 .

[29]  B. Eggum,et al.  Nutritional value of fish viscera silage. , 1981, Journal of the science of food and agriculture.

[30]  Sofia V. Silva,et al.  Caseins as source of bioactive peptides , 2005 .

[31]  G. Vincendon,et al.  Gas-liquid chromatography of the N(O)-heptafluorobutyrates of the isoamyl esters of amino acids , 1973 .

[32]  S. Benjakul,et al.  Protein Hydrolysates from Pacific Whiting Solid Wastes , 1997 .

[33]  W. Verstraete,et al.  A quantitative in silico analysis calculates the angiotensin I converting enzyme (ACE) inhibitory activity in pea and whey protein digests. , 2004, Biochimie.

[34]  K. Kida,et al.  Production of angiotensin I converting enzyme inhibitory peptides from sea bream scales , 2004 .

[35]  A. Lipkowski,et al.  New antihypertensive peptides isolated from rapeseed , 2003, Peptides.

[36]  C. Hyun,et al.  Utilization of bovine blood plasma proteins for the production of angiotensin I converting enzyme inhibitory peptides , 2000 .

[37]  S. Imayasu,et al.  Structure and activity of angiotensin I converting enzyme inhibitory peptides from sake and sake lees. , 1994, Bioscience, biotechnology, and biochemistry.

[38]  S. Ishikawa,et al.  Peptide inhibitors for angiotensin I-converting enzyme from enzymatic hydrolysates of porcine skeletal muscle proteins. , 2001, Meat science.

[39]  Xiu-Lan Chen,et al.  High throughput and rapid screening of marine protein hydrolysates enriched in peptides with angiotensin-I-converting enzyme inhibitory activity by capillary electrophoresis. , 2007, Bioresource technology.

[40]  S. Ferreira,et al.  Isolation of bradykinin-potentiating peptides from Bothrops jararaca venom. , 1970, Biochemistry.

[41]  J. Synowiecki,et al.  Production and characteristics of protein hydrolysates from capelin (Mallotus villosus) , 1995 .

[42]  L. Miclo,et al.  Angiotensin‐I‐converting enzyme inhibitory peptides from tryptic hydrolysate of bovine αS2‐casein , 2002 .

[43]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[44]  M. Nasri,et al.  Biochemical and molecular characterization of a detergent stable alkaline serine-protease from a newly isolated Bacillus licheniformis NH1 , 2007 .