ISOLATION OF HEN EGG WHITE LYSOZYME BY CATION EXCHANGE CHROMATOGRAPHY, ANALYSIS OF ITS DIGESTIBILITY AND EVALUATION OF THE INHIBITION LIPID PEROXIDATION IN THE ZEBRAFISH MODEL

Objective: The aim of this study was to separate and identify lysozyme using cation exchange chromatography, evaluate the protein digestibility andanalyze the inhibition lipid peroxidation in the zebrafish model.Methods: Hen egg white lysozyme was isolate with cation exchange chromatography. Residual muramidase activity was evaluated with thespectrophotometric method. Isolate lysozyme (ILZ) and hydrolysates were analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) electrophoresis and reversed-phase high-performance liquid chromatography (RP-HPLC).Results: To identify the protein, the sample was isolated with cationic exchange chromatography, sample was analyzed using RP-HPLC and SDS-PAGEconfirming that the fraction was hen lysozyme egg white. The enzymatic activity of the isolated protein was normal compared to the commerciallysozyme activity. Hydrolysates had no muramidase activity and were able to inhibit lipid peroxidation in zebrafish larvae.Conclusions: Cation exchange chromatography is a good method to ILZ from egg white. Hydrolysates of lysozyme were effective to inhibit lipidperoxidation in zebrafish model.Keywords: Lysozyme, Cation exchange chromatography, Enzymatic hydrolysis, Muramidase activity

ISOLATION OF PROTEINS FROM SACHA INCHI (PLUKENETIA VOLUBILIS L.) IN PRESENCE OF WATER AND SALT

Objective: The aim of this study was to obtain protein isolate from sacha inchi using alkaline pH at different pHs of precipitation with water and saltand to analyze protein isolate with electrophoresis.Methods: Sacha inchi protein isolates were obtained using isoelectric precipitation method at different pHs. Proteins were analyzed usingelectrophoresis native-polyacrylamide gel electrophoresis (PAGE), one-dimensional, two-dimensional-sodium dodecyl sulfate-PAGE.Results: A yield of 20.88% of protein isolate of defatted sacha inchi flour at pH 4.0 with a 75.31% of protein was obtained. The yield of protein isolateusing water and salt was similar. Polypeptides profile is between 14 and 70 kDa.Conclusions: Sacha inchi seed is a good source of proteins. Globulins and albumins were identified in the sacha inchi protein isolate in the presenceof water and salt.Keywords: Albumins, Globulins, Proteins, Protein isolate, Sacha inchi

ANALYSIS OF PROTEIN ISOLATE FROM QUINOA (CHENOPODIUM QUINOA WILLD)

ABSTRACTObjective: The aim of this study was to obtain protein isolate from quinoa using alkaline pH at different pHs of precipitation and to analyze proteinisolate with electrophoresis.Methods: Quinoa protein isolates were obtained using isoelectric precipitation method at different pHs. Proteins were analyzed using electrophoresisnative-polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate - PAGE.Results: A yield of 6.29% of protein isolate of defatted quinoa at pH 4.0 was obtained. The content of protein isolate was higher than 64% in allpH assays. Globulins and albumins in protein isolate at different pHs were observed. One band near 130 kDa was found. A band with MW 60 kDacorresponding to 7S globulin was found. The bands, MW 33-36 kDa and MW 20-22 kDa, correspond to 11S globulin. Bands less to 15.4 kDa correspondto albumins.Conclusions: Quinoa is a good source of proteins. Globulins and albumins were identified in the quinoa protein isolate.Keywords: Quinoa, Globulins, Albumins, Polypeptides, Protein isolate

ANALYSIS OF SESAME PROTEINS ISOLATE (SESAMUM INDICUM L) WITH WATER AND SALT TREATMENT

Objective: The aim of this study was to obtain protein isolate from sesame using alkaline pH at different pHs of precipitation with water and salt andto analyze protein isolate with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).Methods: Sesame protein isolates were obtained using isoelectric precipitation method at different pHs using water and salt as solvents. Proteinswere analyzed using native-PAGE and SDS-PAGE.Results: A yield of 14,727% ± 0.3 of protein isolate of defatted sesame flour at pH 7.0 with a 47.4% ± 0.6 of protein was obtained. The yield of proteinisolate using water and salt was similar. Polypeptides profile is between 6.5 and 50 kDa.Conclusions: Sesame seed is a good source of proteins. Globulins and albumins were identified in the sesame protein isolate in the presence of waterand salt.Keywords: Sesame, Protein isolate, Proteins, Globulins and albumins

DIGESTIBILITY AND ANTI-INFLAMMATORY ACTIVITY IN VITRO OF SACHA INCHI (PLUKENETIA VOLUBILIS L.) PROTEINS

Objective: The aim of this study was to evaluate the digestibility and anti-inflammatory activity in vitro of sacha inchi protein isolate.Methods: Proteins were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gastric digestibility was evaluated with pepsinat different pHs and at different relation enzyme/substrate. Anti-inflammatory activity in vitro of sacha inchi protein isolate was evaluated usingdenatured protein method with egg albumin.Results: A yield of 20.88% of protein isolate of defatted sacha inchi flour at pH 4.0 with a 93.1% of protein was obtained. 11S globulins were resistantto gastric and duodenal digestion. Sacha inchi protein isolate at pH 4.0 (1000 µg/ml) presents 78.13% of anti-inflammatory activity in vitro.Conclusions: Sacha inchi seed is a good source of proteins. 11S globulins are resistant to pepsin and pancreatin hydrolysis. Sacha inchi protein isolatehas high anti-inflammatory activity in vitro.Keywords: Sacha inchi, Gastric digestion, Duodenal digestion, Globulins and albumins

IDENTIFICATION OF PROTEINS ISOLATE FROM AMARANTH (AMARANTHUS CAUDATUS) BY SODIUM DODECYL SULFATE-POLYACRYLAMIDE GEL ELECTROPHORESIS WITH WATER AND NACL 0.1 M SOLVENTS

ABSTRACTObjective: The aim of this study was to obtain protein isolate from amaranth using alkaline method at extraction pH 8 and extraction pH 12 withdifferent precipitation pHs and to analyze protein isolate with electrophoresis.Methods: Amaranth protein isolates were obtained using isoelectric precipitation method at different pHs. Proteins were analyzed using sodiumdodecyl sulfate-polyacrylamide gel electrophoresis.Results: A yield of 20.52% of protein isolate of defatted amaranth at pH 4.0 was obtained. The content of protein isolate was higher than 53% in allpH assays. Globulins and albumins in protein isolate at different pHs were observed. A band with 50 kDa corresponding to 7S globulin was found. Thebands 36-38 kDa and 18-20 kDa correspond to 11S globulin. Bands less to 14.4 kDa correspond to albumins.Conclusions: Amaranth protein isolate is possible to obtain in extreme conditions of pH. The treatment with water was optimum to obtain amaranthprotein isolate.Keywords: Amaranth, Globulins, Albumins, Proteinisolate, Sodium dodecyl sulfate-polyacrylamide gel electrophoresis