Purification of a Novel Oligophosphopeptide with High Calcium Binding Activity from Carp Egg Hydrolysate

Abstract

Keywords: purification, carp eggs, hydrolysate, dephosphorization, calcium binding activity, oligophosphopeptide IntroductionThe global fishery production reached 178 million tons in 2011(i), more than 50% of which was discarded as inedible byproducts such as bone, skin, internal organs, head and eggs (Jung et al., 2005). Plentiful research have been implemented on utilizing fishery byproducts (Chakraborty et al., 2011;Duan et al., 2009;Kongsri et al., 2013;Mori et al., 2013). However, the utilization studies on eggs, especially on freshwater fish eggs, are very scarce.Eggs are rich in lipids (especially phospholipids and n-3 polyunsaturated fatty acids), the yolk protein, mineral elements, vitamins, carotenoids, and other nutrients. But in the conventional process, eggs are often treated as waste to be abandoned. AbbreviationsCPP; casein phosphopeptide, Hy; carp egg tryptic hydrolysate, DP0; carp eggs without dephosphorization, DP1, DP2, DP3, and DP4; dephosphorized carp eggs with degree of dephosphorization of 13.66%, 30.39%, 48.71%, and 83.49% respectively, U1 and U2; the fractions after ultrafiltration with molecular weight <3 kDa and > 3 kDa respectively, H1, H2 and H3; the fractions eluted with 100 mM, 200 mM and 400 mM phosphate buffer respectively on hydroxyapatite chromatography column, HA; hydroxyapatite chromatography H. Huang et al. 800 Calcium is an important mineral element to maintain human health, which plays an important role in the formation of bones and teeth, neurotransmitter release, muscle composition, and heart beat regulation. Calcium is mainly ingested through drinks and food, especially milk or cheese products with high calcium content. Influenced by lactose intolerance, dietary habits, and other factors,Asians intake less milk and cheese products and their food mainly focuses on vegetable diet. However, substances such as the phytic acid, oxalic acid, saturated fatty acids, and dietary fiber are easily combined with calcium to form insoluble matter, which is not beneficial to calcium absorption.Many peptides isolated from animals possessed the ability of binding Ca Lee et al., 2009;Jiang et al., 2000;Jung et al., 2007). Casein phosphopeptides (CPPs) were well known for their metal binding abilities because the CPPs contained electronegative phosphoserines (West et al., 1986;Meisel et al., 1990;Yuan et al., 1991;Gerber et al., 1986;Sato et al., 1986). The phosphopeptides obtained from the enzymolysis of fish bone (Jung et al., 2007(Jung et al., , 2006(Jung et al., , 2005 can also chelate Ca, whose performance is similar to CPPs. Many studies indicated that fish eggs also contain high phosphoprotein and can obtain phosphopeptides by hydrolysis (Catherine et al., 2002;Inoue et al., 1971).Carp was one of the most productive (over 3.8 million tons ) freshwater fish species in the world (ii). In breeding season, the ovarian can weigh 10% to 25.81% of the overall weight. In the present study, we will discuss the appropriate degree of dephosphorization of carp eggs before trypsin hydrolysi...

Identification of a new calcium‐binding peptide from enzymatic proteolysate of Acetes japonicus

“…It was shown that the <1 kDa fraction had the highest activity (277.96 mg Ca 2+ /g protein), which was lower than that of CPP about 1.7 folds. This finding was in agreement with those of Huang et al () and Huang et al () who reported that the lowest molecular weight peptide showed the highest calcium‐binding activity.…”

Bioactive Peptides Isolated from Casein Phosphopeptides Enhance Calcium and Magnesium Uptake in Caco-2 Cell Monolayers

“…7 However, numerous studies have focused on using organic supplements due to their increased bioavailability. 8−10 To this end, many organic supplements improve the absorptive ability of mineral elements and include oligophosphopeptide, 11 chicken eggshell matrix proteins, 12 tilapia protein hydrolysate, 13 whey protein hydrolysates, 14 hen egg yolk phosvitin, 15 and desalted duck egg white peptides. 16 Casein phosphopeptides (CPPs) are derived from trypsinmediated casein cleavage and are considered mineral carriers with potential roles in improving elemental mineral absorption.…”

Purification of Bovine Bone Oligophosphopeptide with High Calcium-binding Activity by <i>Bacillus cereus </i>MBL13 Collagenolytic Protease

“…As shown in Figure 3 (c), two major fractions (R1, R2) were separated through the semi-preparative C18 column. The calcium-binding activity of an oligophosphopeptide from carp egg hydrolysate and pollack backbone peptide were 5.0, 7.0, and 7.85 mmol/g-protein when the protein concentration was 50 mg/L [6,7, 28,35]. Compared with the peptides above, the peptide derived from bovine bone showed that R2 (BBP) had the highest calcium-binding activity of 8.25 mmol/g-protein.…”

“…H4 was eluted with the maximum concentration of phosphate buffer (150 mM) and exhibited the highest calcium-binding ability of 7.02 mmol/gprotein, which showed that HA chromatography was very effective in separating peptide with the calcium-binding activity. The oligophosphopeptides with calcium-binding activity have been successfully separated and obtained from fish bone and carp egg [7,28] using HA affinity chromatography.…”

“…The assay was performed according to the method of Jung and Huang with the following modifications [6,28]. The various concentrations of hydrolysates and BBP of up to 500 mg/L were mixed with 5 mM of CaCl 2 and 20 mM of sodium phosphate buffer (pH 7.5).…”