Isolation and Characterization of Antioxidative Peptides from Gelatin Hydrolysate of Alaska Pollack Skin

Abstract

Gelatin extracted from Alaska pollack skin was hydrolyzed with serial digestions in the order of Alcalase, Pronase E, and collagenase using a three-step recycling membrane reactor. The fraction from the second step, which was hydrolyzed with Pronase E, was composed of peptides ranging from 1.5 to 4.5 kDa and showed high antioxidative activity. Two different peptides showing strong antioxidative activity were isolated from the hydrolysate using consecutive chromatographic methods including gel filtration on a Sephadex G-25 column, ion-exchange chromatography on a SP-Sephadex C-25 column, and high-performance liquid chromatography on an ODS column. The isolated peptides, P1 and P2, were composed of 13 and 16 amino acid residues, respectively; and both peptides contained a Gly residue at the C-terminus and the repeating motif Gly-Pro-Hyp. The antioxidative activities of the purified peptides were measured using the thiobarbituric acid method, and the cell viability was measured with MTT assay. The results showed that P2 had potent antioxidative activity on peroxidation of linoleic acid. Moreover, the cell viability of cultured liver cells was significantly enhanced by addition of the peptide. These results indicate that the purified peptide, P2, from gelatin hydrolysate of Alaska pollack skin is a natural antioxidant which has potent antioxidative activity.

Development of fish protein powder as an ingredient for food applications: a review

“…3 pH-shift process for production fish protein isolates (FPI) (adapted from Hultin et al 2005) are influenced by the degree of hydrolysis (Kristinsson and Rasco 2000). Proteins separated by partial enzyme hydrolysis are known to possess anti-oxidation properties against peroxidation of lipids and fatty acids (Kim et al 2001). …”

Fractionation and Characterization of Fractions with High Antioxidative Activity from the Gelatin Hydrolysates of Korean Rockfish Sebastes schlegelii Skin

“…Although several studies have examined the fractionation and isolation of functional peptides from hydrolysates of fish skin (Kim et al, 2001a(Kim et al, , 2001bMendis et al, 2005), with the exception of Kim et al (2011), no research has evaluated the feasibility of using rockfish skin gelatin as a source of functional gelatin hydrolysates. The objective of this study was to obtain a fraction with high antioxidant activity from rockfish skin gelatin hydrolysates by fractionation through UF membranes.…”

“…Mendis et al (2005) reported that antioxidant peptides isolated from hoki skin gelatin hydrolysate were composed of His-Gly-Pro-LeuGly-Pro-Leu. Kim et al (2001b) reported that the peptides (P1 and P2) fractionated and isolated from gelatin hydrolysate of Alaska pollock skin were composed of 13 and 16 amino acid residues, respectively, and that both peptides contained glycine residues at the C-terminus and repeating glycine-prolinehydroxyproline. These reports, as well as our results, suggest that sequential enzymatic hydrolysis of rockfish skin gelatin with Alcalase/Flavourzyme followed by fractionation are suitable methods for obtaining antioxidant peptides.…”

“…The intact form of the gelatin contains many bioactive proteins due to the presence of biologically active peptide sequences in their primary structures. Numerous methods have been used to release bioactive peptide fractions from native proteins; proteolytic digestion and fractionation through ultrafiltration (UF) membranes with different molecular weight cut-offs (MWCOs) is one of the most common methods (Kim et al, 1994(Kim et al, , 2001bJeon et al, 2000). Fractionation through UF membranes has several advantages, such as the mass-production capability and the enhancement of some functional properties in comparison with the original hydrolysate.…”

Optimization of goldstripe sardine (Sardinella gibbosa) protein hydrolysate using Alcalase® 2.4L by response surface methodology Optimización de hidrolisato de proteína de Sardinela dorada (Sardinella gibbosa) usando Alcalase® 2.4L a través de RSM

“…Microbial enzymes, especially Alcalase 1 2.4L (from Bacillus licheniformis), have been repeatedly favored for fish hydrolysis due to the high degree of hydrolysis (DH) that can be achieved, compared to neutral or acidic enzymes, in a relatively short time under moderate conditions (Je, Park, & Kim, 2004;Kim, Kim, Byun, Joo, & Shahidi, 2001;Rajapakse, Mendis, Byun, & Kim, 2005;Shahidi & Amarowicz, 1996).…”