• Preventive Nutrition and Food Science
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• v.15 no.4
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• pp.282-286
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• 2010

To isolate a calcium-binding peptide from chlorella protein hydrolysates, chlorella protein was extracted and hydrolyzed using Flavourzyme, a commercial protease. The degree of hydrolysis and calcium-binding capacity were determined using trinitrobenzenesulfonic acid and orthophenanthroline methods, respectively. The enzymatic hydrolysis of chlorella protein for 6 hr was sufficient for the preparation of chlorella protein hydrolysates. The hydrolysates of chlorella protein were then ultra-filtered under 5 kDa as molecular weight. The membrane-filtered solution was fractionated using ion exchange, reverse phase, normal phase chromatography, and fast protein liquid chromatography to identify a calcium-binding peptide. The purified calcium-binding peptide had a calcium binding activity of 0.166 mM and was determined to be 700.48 Da as molecular weight, and partially identified as a peptide containing Asn-Ser-Gly-Cys based on liquid chromatography/electrospray ionization tandem mass spectrum.

• Journal of Applied Biological Chemistry
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• v.53 no.3
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• pp.174-178
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• 2010

Calcium and iron binding peptides were prepared by enzymatic hydrolysis and ultrafiltration of rice bran protein (RBP), which was isolated from defatted rice bran by phytase and xylanase treatment and ultrasonication. The isolated RBP had a molecular weight in the range of 10-66 kDa. The extracted proteins were hydrolyzed using Flavourzyme for 6 hr. After ultrafiltration under 5 kDa as molecular weight, the peptides were fractionated into 4 peaks by Sephadex G-15 gel permeation chromatography, and each fraction was determined for calcium and iron binding activity. As the result, Fl and F2 fractions were the best candidate for calcium and iron chelation, respectively. These results suggest that the calcium and iron binding peptides can be used as functional food additives in food industry.

• Journal of the East Asian Society of Dietary Life
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• v.20 no.5
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• pp.680-686
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• 2010

Silk sericin protein was hydrolyzed by seven proteolytic enzymes in order to examine the effectiveness of the hydrolysates in binding calcium. The amino acid nitrogen content of hydrolysates from Flavourzyme was higher than that for other enzymes, and its calcium binding capacity showed a dose-dependent increase. We examined the effects of calcium binding peptide from sericin hydolysates on the bioavailability of Ca-deficient rats. Three-week-old male rats were fed an Ca-deficient diet for three weeks. Rats were divided into four groups (DD: non-treated group on calcium deficient diet; DD+MC: milk-calcium treated group; DD+OC: organic calcium made using sericin hydolysates; and DD+IC: inorganic calcium ($CaCl_2$). After oral administration of calcium supplements for one week, the calcium content of the serum and liver were significantly higher in DD+OC ($101.7{\mu}g$/mL and $49.3{\mu}g$/mL) and DD+MC ($83.6{\mu}g$/mL and $42.8{\mu}g$/mL) than DD ($86.3{\mu}g$/mL and $43.4{\mu}g$/mL). The alkaline phosphatase (ALP) content in the treated groups was significantly lower than DD, but no significant difference among groups was shown. Aspartate aminotransferase (AST) levels did not show any significant difference between groups. Alanine aminotransferase (ALT) levels were significantly reduced compared to the DD group. In conclusion, binding calcium to peptides from sericin hydrolysates seems to improve its bioavailability, and to hasten the cure of calcium deficiency in experimental rats.

• The Korean Journal of Food And Nutrition
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• v.10 no.4
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• pp.485-490
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• 1997

Gluten peptide was produced from corn gluten by enzymatic hydrolysis. This peptide had an ability to increased the solubility of calcium owing to protect calcium ions from forming precipitates of calcium phosphate in the presence of phosphate ions. The solubility of calcium was increased 5.2 times in the presence of 8.3 mg peptide produced by the treatment of papain. These peptides contained high acidic amino acids and fractionated by Delta pack column into fractions No. 1, No. 2 and No. 3. Among them the fraction No. 3 had the highest calcium binding capacity.

• Food Science and Preservation
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• v.22 no.2
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• pp.297-302
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• 2015

Calcium is one of the essential mineral for the humans due to its crucial physiological functions in the body. Calcium deficiency results in many diseases, such as osteoporosis. Therefore, calcium supplements are available as a functional food. However, most calcium supplements in the market have a limitation due to poor absorption and low bioavailability. Thus, calcium-chelated peptides for improving the absorption rate of calcium have been isolated from foods including porcine meat and bone meal (MBM), and mussel using the enzymatic hydrolysis of their protein. The hydrolysates of food were ultra-filtered in order to obtain small peptides less than 3 kDa and the Ca-binding peptides were isolated via the anion exchange chromatography. The binding activity and concentration of Ca-binding pepetides were determined. In particular, the MBM and mussel protein hydrolysates were fractionated by mono Q and Q-Sepharose, respectively. As a result, among the fractions, the fractions of MBM F2 and mussel F3 showed the highest Ca-binding activity. These results suggest that MBM and mussel protein hydrolysates can be used as calcium supplements.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1459-1464
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• 2004

The purpose of this research was to investigate the potential use of cheese whey protein (CWP), a cheese by-product. The physiological activity of calcium-binding peptides in CWP may be used as a food additive that prevents bone disorders. This research also examined the characteristics of calcium-binding peptides. After the CWP was heat treated, it was hydrolyzed by trypsin. Then calcium-binding peptides were separated and purified by ion-exchange chromatography and reverse phase HPLC, respectively. To examine the characteristics of the purified calcium-binding peptides, amino acid composition and amino acid sequence were analyzed. Calcium-binding peptides with a small molecular weight of about 1.4 to 3.4 kDa were identified in the fraction that was flowed out from 0.25 M NaCl step gradient by ion-exchange chromatography of tryptic hydrolysates. The results of the amino acid analysis revealed that glutamic acid in a calcium-binding site took up most part of the amino acids including a quantity of proline, leucine and lysine. The amino acid sequence of calcium-binding peptides showed Phe-Leu-Asp-Asp-Asp-Leu-Thr-Asp and Ile-Leu-Asp-Lys from $\alpha$-LA and Ile-Pro-Ala-Val-Phe-Lys and Val-Tyr-Val-Glu-Glu-Leu-Lys from ${\beta}$-LG.

• Food Science and Preservation
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• v.19 no.3
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• pp.438-442
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• 2012

To prepare calcium-binding peptides as calcium supplement, barley proteins were hydrolyzed using Flavourzyme for 18 h and the hydrolysates were ultra-filtered under 3 kDa as a molecular weight. The resultant filtered peptides were fractionated using ion exchange and normal-phase high performance liquid chromatography. Then each fraction that was obtained was determined for its calcium-binding activity to isolate the calcium-binding peptides. As a result, the highest calcium-binding peptide fraction was obtained, and the results suggest that barley protein hydrolysates can be used as a calcium supplement.

• Journal of Periodontal and Implant Science
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• v.42 no.5
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• pp.166-172
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• 2012

Purpose: The aim of this study was to evaluate the improvement of osteogenic potential of biphasic calcium phosphate (BCP) bone substitute coated with synthetic cell-binding peptide sequences in a standardized rabbit sinus model. Methods: Standardized 6-mm diameter defects were created bilaterally on the maxillary sinus of ten male New Zealand white rabbits, receiving BCP bone substitute coated with synthetic cell binding peptide sequences on one side (experimental group) and BCP bone substitute without coating (control group) on the other side. Histologic and histomorphometric analysis of bone formation was carried out after a healing period of 4 or 8 weeks. Results: Histological analysis revealed signs of new bone formation in both experimental groups (4- and 8-week healing groups) with a statistically significant increase in bone formation in the 4-week healing group compared to the control group. However, no statistically significant difference in bone formation was found between the 8-week healing group and the control group. Conclusions: This study found that BCP bone substitute coated with synthetic cell-binding peptide sequences enhanced osteoinductive potential in a standardized rabbit sinus model and its effectiveness was greater in the 4-week healing group than in the 8-week healing group.

• Journal of Applied Biological Chemistry
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• v.55 no.4
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• pp.263-266
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• 2012

Isolation of iron and calcium-binding peptides derived from cottonseed meal protein (CMP) hydrolysates was investigated. The degree of hydrolysis of CMP by Flavourzyme was monitored using trinitrobenzenesulfonic acid method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Enzymatic hydrolysis of CMP for 12 h was sufficient for the preparation of CMP hydrolysates, and the hydrolysates were membrane-filtered under 3 kDa as a molecular weight. The filtered solution was fractionated using Q-Sepharose fast flow, Sephadex G-15, and reversed phase-high performance liquid chromatography for iron and calcium-binding peptides. As a result, F51 fraction was obtained as the best candidate for calcium and iron chelation, and the isolated iron and calcium-binding peptides can be used as functional food additives, similar to iron and calcium supplements.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.139-146
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• 2001

This study was carry out to identify the distribution of calcium binding proteins; calbindin(CB), calretinin(CR) and parvalbumin(PA) in the suprachiasmatic nucleus(SCN) of the Korean native goat by immunohistochemical methods. The expression of substance P(SP), calcitonin gene-related peptide(CG-RP), neuropeptide Y(NPY), vasoactive intestinal polypeptide(VIP) and galanin(GAL) were also investigated. CR-immunoreactivity was found in both of the cell bodies and fibers in the SCN, which the CB-immunoreactivity was observed only in the fibers. The immunoreactivity for VIP was observed in both the cell bodies and fibers, but SP-, NPY, GAL-immunoreactivities were only found in the fibers. CGRP-immunoreactivity was not seen in cell body and fibers. These results suggest that VIP, SP, NPY and GAL play a neuromodulatory or/ and neurotransmitter roles in cooperation with CB and CR in SCN of the Korean native goat.