| Hyperlipidemia is a common and multiple metabolic disease,which is an important factor inducing cardiovascular and cerebrovascular diseases such as atherosclerosis and coronary heart disease,as well as common metabolic diseases such as obesity and diabetes,seriously endangering human health.Quinoa is a kind of high protein high quality multigrain,protein content in grain is higher,amino acid composition ratio is reasonable,have the biological activity such as oxidation resistance,but the research to its reduce blood fat activity is less.This study quinoa as raw material,extracted quinoa protein,bile salts adsorption as evaluation index,the enzyme hydrolysis of quinoa peptides with cholesterol-lowering activity,and make use of hyperlipidemia mouse model to explore the lipid-lowering effect of quinoa protein and polypeptide,at the same time to quinoa peptides with efficacy as the main raw material to develop a quinoa polypeptide beverage,the main research results are as follows:1.The quinoa protein was extracted by alkali-soluble acid precipitation method.The results showed that the protein content of the extracted quinoa protein was 68.22%±3.19%,(n= 5);Mainly distributed in 50,35-40,30-35,15-25 kDa;amino acid test results showed that the content of essential amino acids,hydrophobic amino acids and arginine in quinoa protein accounted for 38.21%,37.88% and 9.91%.The in vitro cholate adsorption experiment simulated the hypolipidemic activity of quinoa protein,and the results showed that the adsorption of quinoa protein on cholate was fully saturated at 60 min;gradually increased with the addition of quinoa protein and cholate After the enhancement,it tends to be saturated;when the concentration of cholate is 5 mg/mL,the adsorption capacity of 1 g of quinoa protein to cholate is(226.84±0.42)mg.2.Enzymatic hydrolysis of quinoa protein: comprehensive evaluation based on the degree of hydrolysis(DH)and in vitro cholate adsorption as indicators,screening reaction proteases with better enzymatic hydrolysis effects,designing single-factor test and orthogonal test to optimize and get the best The enzymolysis process was used to prepare quinoa polypeptide with hypolipidemic activity.The results showed that the effect of flavor protease enzymolysis is better.The best enzymolysis process is: enzymolysis pH=7.0,enzymolysis temperature 55 ℃,enzyme addition amount 6000 U/g,substrate concentration 3%,enzymolysis time 2 h.Under the optimal enzymatic hydrolysis conditions,the degree of hydrolysis(DH)was 38.22%±0.65%,and the total amount of quinoa polypeptide adsorbed cholate reached(7.56±0.11)mg / mL.The results of amino acid determination showed that the content of hydrophobic amino acids and arginine in quinoa polypeptide accounted for38.72% and 9.44% of the total amino acids,respectively.A total of 472 peptide sequences were identified by quinoa polypeptide mass spectrometry,among which 13 peptides withmolecular weight around 1000 Da and the peptide end residue as arginine(R)were screened out,which may be an important factor in its anti-lipid activity in vitro.The results showed that it was mainly derived from 11 S seed storage globulin with molecular weights of 53.576 kDa and 153.668 kDa,and n-glycosidase proteins with molecular weights of 31.377 kDa and31.419 kDa,respectively.3.The high-fat mouse model was divided into a blank control group,a high-fat model group,a positive drug group,a high-dose quinoa peptide group,a low-dose quinoa peptide group,a quinoa proteome group,and the rats were administrated with quinoa protein and peptide to intervene After 30 days of gavage,the effects of quinoa protein and peptide on the body condition,organ index,blood lipid level and liver function of mice on a high-fat diet were analyzed.The results showed that high-fat feeding significantly increased mouse body weight,liver index,and fat coefficient.Gavage of quinoa proteins and peptides significantly inhibited food utilization,weight gain,liver coefficient,and fat coefficient of hyperlipidemia mice.Effect,it has no significant effect on the heart,kidney,spleen and other organs of experimental mice;gavage of quinoa protein and peptide can significantly reduce the serum total cholesterol(TC)level of high-fat-fed mice,and increase high-density lipoproteinCholesterol(HDL-C)levels also have a certain downward trend in serum low-density lipoprotein-cholesterol(LDL-C)levels,which may have no effect on serum total triglyceride(TG)levels,and the quinoa peptide group is small Rats are more effective than mice fed with quinoa protein;they can significantly reduce total liver cholesterol(TC)levels and increase total bile acid levels in the liver;they can significantly reduce alanine aminotransferase(ALT)and aspartate aminotransferase in the serum of high-fat fed mice(AST)activity;the results of pathological section show that high dose of quinoa polypeptide can alleviate liver damage and restore its pathological state.4.Study on the technology of quinoa peptide beverage: single factor experiment to investigate the addition of beverage to water ratio,sweetener(sucrose,inulin,maltitol),sour agent(citric acid)and flavoring agent(β-cyclodextrin)The comprehensive evaluation using fuzzy mathematical evaluation model,the design of orthogonal experiments,the determination of the beverage-to-water ratio and the amount of each auxiliary;the Zeta potential and centrifugal sedimentation rate were used as evaluation indicators to investigate the beverage stabilizer(xanthan gum,seaweed Acid propylene glycol ester);analysis of the sensory,physical and chemical,microbiological,and in vitro bile acid salt digestion activities of the beverage.The results showed that the optimal ratio of beverage was 4: 6 ratio of material to water,6.0%(W/V)of sucrose,2.0%(W/V)of maltitol,and 3.0%(W/V)of inulin,0.02%(W/V)citric acid,0.1%(W/V)β-cyclodextrin;propylene glycol alginate as thestabilizer,0.05%(W/V);The beverage has good sensory quality and stable physical and chemical properties.After simulated gastric and intestinal digestion in vitro,it has a good adsorption effect on bile salts,and the total adsorption is(3.76±0.21)mg/mL.The above experimental results showed that quinoa protein and polypeptide had better adsorption of cholate,indicating its in vitro hypolipidemic activity was better;Secondly,the molecular weight of quinoa polypeptide is about 1000 Da peptide terminal residues for arginine peptide segment may be its in vitro lipid lowering activity is better important factors,its main source may be quinoa protein 11 S seed storage globulin and N-glycosidase protein;Lavage quinoa protein and polypeptide on hyperlipemia mice,coefficient of weight gain,liver,and fat food utilization coefficient has significant inhibitory effect,can adjust the blood lipid level of mice a high-fat feeding,promote the decomposition of the liver cholesterol,improve the hyperlipemia mice liver injury,the callback is the pathological state,to prevent hyperlipidemia has the potential effect;At the same time,a nutritious,delicious,resistant quinoa polypeptide drink was developed to enrich the diversity of quinoa products. |
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