the hydrolysates protu00c9iques goby (zosterisessor ophiocephalus) with hypotensive activity

摘要

the present invention describes the preparation of protein hydrolysates (z. ophiocephalus) muscle of round, rich in bioactive peptides by the action of proteolytic enzymes on microbial and, more particularly, the invention describes the splitting of the hydrolysate and the characterization of fractions were obtained.according to the method of the invention, the production of peptide inhibitors of the angiotensin converting enzyme (ace) is based on a controlled enzymatic hydrolysis of protein in the muscle of round under the action of exogenous proteases, followed by systematic methods of fractionation, biological tests.the optimization of the proteolytic enzyme was used to determine the enzyme preparation and the degree of hydrolysis (dh) of the protein hydrolysate with the highest activity. the anti ace activity of the hydrolysates prepared was studied by using the histidin hhl (hippuryl - (e) - leucine as substrate.the inhibitory activity of the hydrolysates were treated with tca and protein hydrolysate preparation bacillus mojavensis a21 proteolytic this antihypertensive activity is greater for a 67,95% dh about 13.21% and a concentration of 2.5 mg / ml).the protein hydrolysate having a dh of 11.07% has the highest activity, obtained from the preparation of a21 was further fractionated on a column of sephadex g-25 gel filtration fractions in the five majority (f1 - f5). this fraction f5 to ace inhibition (ic50 = 0.32, the most important (0.01 mg / ml).the fraction f was subjected to further fractionation by rp - hplc with c18 reversed-phase silica column. the profile has a number of peaks, indicating the abundance of peptides generated. four fractions were collected and concentrated by speed vac.the test of anti ace activity showed that all the fractions are active and that the highest activity was obtained with the fraction f5 / 2 (99% at 0.5 mg / ml). the latter has an ic50 of about 0,1u00b10,02 mg / ml).analysis of the active fractions by esi-ms / ms showed that the sub fraction f5 2 contains five peptides: ala arg ser, val val ala pro phe ala and gly thr, arg ser thr ala, phe tyr pro pro, arg cys ser ala gly val, 15 peptides have been identified in the g1 fraction 4.