The Study Of Pepper Meal Protein Enzymolysis To Produce Flavor Peptide

Pepper meal, as a by-product left behind after the extraction of capsanthin and capsaicin from red chili, is generally slightly red or light yellow powder. At present, the processing of hot pepper meal processing is generally sold at a low price to farmers, or as a waste directly discarded, can not fully use the residual nutrients in pepper meal, resulting in a great waste of resources. The nutrition analysis of pepper meal which bought on the market shows that it contains crude fat 1.91%, total sugar 4.30%,10.88% of moisture, ash 9.56%, and the content of cellulose up 31.95%, protein content can reach higher 15.75%. Therefore, the pepper meal is a potential, ideal for high quality protein resources and the raw material of flavor peptide. Based on pepper meal protein (protein purity of about 59.14%.) as raw materials made in the laboratory, flavor peptide constituent was obtained according to the enzymatic hydrolysis and ultrafiltration separation, gel filtration chromatography and RP-HPLC purification.The main results were as follows:According to the degree of hydrolysis, peptide to rate and sensory evalution as the index, trypsin, papain, alcalase, protamex 4 kinds of protease from screened hydrolysis pepper meal protein was the best enzyme for composite protease. Research of solid-liquid ratio, enzyme dosage, time, pH and temperature on the enzymolysis effect, the optimum solid-liquid ratio was determined as 1:10 (m:V), by single factor test and orthogonal test optimization test, the optimum technological conditions on the protamex protease single enzyme hydrolysis of pepper meal protein, were that the solid-liquid ratio was 1:10 (m:V), enzyme dosage 2.0%, enzymolysis time 4h, pH 6.5 and temperature of 50?. Under these conditions, the hydrolysis degree was 19.60% and the yield of peptide was 6.30%, the umami score value reached 6.0, comprehensive flavor was 2.8.Because the hydrolysis rate of plant protein is lower than that of animal protein, the single enzyme hydrolysis is difficult to obtain enough short peptide. The effects of mixed ratio, total enzyme dosage, time, pH and temperature on the enzymatic hydrolysis were studied in the experiment of double enzyme (protamex and flavorzyme) hydrolysis pepper meal protein. The result was that the best total enzyme dosage was 2.0%. By single factor and orthogonal experiments, the optimum technological conditions on protamex and flavorzyme double enzyme hydrolysis pepper meal protein were protamex and flavourzyme quality compound ratio of 2:1, total enzyme dosage 2.0%, hydrolysis time of 6 h, pH 6.5, temperature 50?. Under above conditions, the degree of hydrolysis was 19.44%, the yield of peptide was 6.52%, the umami score value reached 7.0, comprehensive flavor was 3.4. Mixed analysis of variance of the enzyme ratio, enzymolysis pH factors at 0.01 level significantly affected. Comprehensive comparison, and ultimately determine the enzyme solution, the optimum conditions of pepper meal protein:solid-liquid ratio was 1:10 (m:V), protamex and flavorzyme quality compound ratio is 2:1, total enzyme dosage is 2.0%, hydrolysis time of 6 h, pH 6.5, temperature 50?.Study with a ultrafiltration membrane of 3kDa on the optimum enzyme solution process under the condition of the hydrolysate of ultrafiltration were obtained group I (less than 3KDa) and II (above 3KDa) and ultrafiltrates were collected after, then the sensory evaluation on this group of two components was performed. The results showed that the sensory evaluation of group I was better flavor than that of group II, and the umami score obviously higher. The gel chromatography was carried out on the filtrate fraction I, and the better flavor of the peptide obtained from the gel SephadexG-15 than that from SephadexG-25. Selecting the SephadexG-15 gel column, sample volume of 1mL and eluent ultrapure water of condition on the concentration of 25mg/mL, elutionflow rate of 30mL/h, the isolated were obtained three-eluting peaks(A?B?C) and the second component I B tasted better, namely the umami score value reached 7.0, comprehensive flavor was 3.5, the molecule weight 307.32Da-1422.69Da.The I B peak's sample was separated by RP-HPLC method. The gradient elution conditions of mobile phase was decided. The results showed that mobile phase was ultrapure water-acetonitrile (40:60; volume ratio) and the peptide of I B b peak tasted better, the umami score value reached 8.0, comprehensive flavor was 4.0.