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Research on Optimizing Technological Parameter of Enzymatic Preparation of Antihypertensive Peptides Derived from Swine Hemoglobin and Hypertensive Activities in Vitro

Author: GuoQiLiang
Tutor: LiCheng
School: Sichuan Agricultural University
Course: Agricultural Products Processing and Storage
Keywords: Pig hemoglobin Trypsin Enzymatic process parameters Hypotensive activity
CLC: TQ464.7
Type: Master's thesis
Year: 2010
Downloads: 187
Quote: 1
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Abstract


The pig hemoglobin produced by proteolysis hypotensive activity polypeptide is a food-borne bioactive peptides, high security, research reported in the step-down functions with outstanding performance expected as a buck a natural component widely used. In this study, angiotensin-converting enzyme (Angiotensin I-Converting Enzyme, ACE) inhibition rate indicators, compared hydrolysates obtained by the the different proteolytic pig hemoglobin blood pressure lowering activity (ie ACE inhibitory activity), and the hydrolysates hypotensive activity filtering for a optimum hydrolysis with enzymes. Selection of trypsin, pepsin and papain as hydrolysis with enzymes, determination of the three protease enzymatic pig hemoglobin products in each suitable for enzymatic hydrolysis conditions under hypotensive activity and its degree of hydrolysis of porcine hemoglobin as substrate. The research results show that three kinds of enzymatic hydrolysis experiments the different proteolytic six hours after the degree sequence of papain gt; the pepsin gt; trypsin, and ACE inhibition rate pepsin gt; trypsin gt; papain. Screened trypsin as a hydrolysis with an enzyme prepared having a higher hypotensive activity of porcine hemoglobin hydrolysates. Trypsin as a hydrolytic enzyme, substrate concentration, hydrolysis temperature, pH value, enzyme dosage and hydrolysis time optimal the enzymolysis level range for hydrolysis, evaluation of each factor under single factor experiments on pig hemoglobin, and by design orthogonal time to optimize the process parameters on the enzymatic hydrolysis, and enzymatic degradation products have higher blood pressure lowering activity. The results showed that the optimum conditions for trypsin for 10% of the substrate concentration, hydrolysis temperature 40-C, pH value of 9.0, hydrolysis time 6h and the enzyme dosage 2000U / g, the product of ACE inhibition rate was 62.54%. As trypsin hydrolysis with enzymes, optimize the optimum conditions for pig hemoglobin digestion, enzymatic degradation products will be prepared the ultrafiltration separation, in order to examine the enzymatic degradation products of different molecular weight range of blood pressure lowering activity. The results show that each component ACE inhibitory activity of the size of the order of: Lc (Molecular Weight lt; 5KDa) gt; the Oc (Enzymolysis dope) gt; Mc (10 kDa gt; molecular weight gt; 5KDa) gt; Hc (molecular weight greater than 10 kDa), ACE inhibition of the most active group is divided into the component of a molecular weight less than 5 kDa. A molecular weight of less than 5 kDa component formulated as 4,8,12,16,20,24,28 μ / mL 7 different concentrations of the solution, Karp Toledo is formulated as 0.4,0.8,1.2,1.6,2.0,2.4. 2.8ng/mL7 species concentration in the solution, and samples were measured ACE inhibition rate. The results show that the the pig hemoglobin enzymatic molecular weight less than 5KDa component of ACE inhibitory activity IC50 13.69μg/mL, strong blood pressure lowering activity in vitro. Control samples IC50 was in line with IC50 values ??reported Karp Toledo instructions ACE inhibitory activity of the enzymatic degradation products of the experiment is feasible, and the description of the experimental determination of the sample ACE inhibition rate scientific. PREPARATIONS pepsin, pepsin, chymotrypsin, pepsin, chymotrypsin trypsin three kinds of simulated gastrointestinal digestion environment, without the two components of the ultrafiltration and the ultrafiltration treatment further digestion process was investigated The hydrolysates In digestive enzyme treatment before and after the change, its ACE inhibitory judge gastrointestinal digestion stability as the evaluation index. The results show that: the molecular weight is less than 5 kDa component in the the treated ACE inhibitory rate did not change significantly. Enzymatic dope pig blood polypeptide by processing, the ACE inhibitory activity are significantly reduced. According to the results assessed by trypsin digestion pig hemoglobin prepared antihypertensive peptides molecular weight less than 5KDa components with high gastrointestinal digestion stability. The experiments of pig's blood polypeptide as hypotensive activity products for industrial production experimental basis, provide an experimental basis for the future development of pig's blood to blood pressure lowering activity of peptide. Also explore a new way for the deep processing of pig's blood.

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CLC: > Industrial Technology > Chemical Industry > Pharmaceutical chemical industry > Drug production of biological products > Amino acids,peptides,proteins
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