• Preventive Nutrition and Food Science
• /
• 제15권4호
• /
• pp.282-286
• /
• 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
• /
• 제53권3호
• /
• pp.174-178
• /
• 2010

탈지 미강으로부터 미강단백질을 추출하고 상업용 단백분해 효소로 가수분해하고 한외여과하여 얻어진 미강단백질 가수분해물을 Sephadex G-15로 분리하여 얻어진 peptide fraction에 칼슘, 철분을 binding하여 칼슘, 철분 함유 peptide를 제조하였다. 추출된 탈지 미강 단백질의 분자량은 10~66 kDa에 분포하고 있었다. 추출된 단백질을 Flavourzyme으로 가수분해 시, 최적 가수분해 시간은 6시간이었으며, 5kDa 이하로 한외여과 하여 얻어진 peptide를 Sephadex G-15로 분획한 결과 4개의 major peak를 얻었는데, 각 fraction의 칼슘, 철분을 binding한 결과 Ca/peptide는 FI에서, Fe/peptide는 F2에서 가장 많은 함량을 나타내었다. 본 연구 결과 얻어진 칼슘, 철분 binding peptide는 biomineral 기능성 식품의 소재로써 식품산업에 활용될 수 있다고 판단된다.

• 동아시아식생활학회지
• /
• 제20권5호
• /
• pp.680-686
• /
• 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.

• 한국식품영양학회지
• /
• 제10권4호
• /
• pp.485-490
• /
• 1997

옥수수 글루텐을 식물성 효소인 파파인으로 가수분해하여 글루텐 펩타이드를 생산하였다. 생산된 글루펜 펩타이드류는 인이온 존재하에서 인이온과 칼슘이온과의 침전형성을 방지하여 칼슘의 용해성을 증진시켰다. 칼슘이온의 용해도는 8.3mg의 펩타이드 존재시 대조구에 비해 5.2배 증대되었다. 산성 아미노산의 함량이 매우 높은 이들 펩타이드류를 Delta pak 칼럼을 이용하여 분획하였으며 이들 분획 가운데 3번째 분획이 가장 높은 칼슘 용해성을 보였다.

• 한국식품저장유통학회지
• /
• 제22권2호
• /
• pp.297-302
• /
• 2015

저활용 단백질로부터 칼슘 결합물질을 분리하기 위해 돼지 육골분과 진주담치 단백질을 단백질 분해 효소인 alcalase를 이용하여 가수분해물을 제조하였고, 체내 흡수가 용이한 3 kDa 이하로 한외여과 하였다. 돼지 육골분 가수분해물은 Mono Q 컬럼을 통해 분리하였고, 진주담치 가수분해물의 경우 Q-Sepharose로 분리 하여 각각 2개, 3개의 peptide fraction을 얻어 각 fraction의 칼슘 결합력을 측정하였다. 그 결과 MBM F2와 Mussel F3에서 가장 높은 칼슘 결합력을 나타내었고, 따라서 본 연구 결과로 얻어진 가수분해물들은 칼슘 보충 소재로 활용될 수 있다고 판단된다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제17권10호
• /
• pp.1459-1464
• /
• 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.

• 한국식품저장유통학회지
• /
• 제19권3호
• /
• pp.438-442
• /
• 2012

칼슘보충제로 칼슘 결합 펩타이드를 만들기 위해, 보리 단백질에 Flavourzyme을 사용하여 18시간 동안 가수분해 하였고, 가수분해 된 보리 단백질은 3 kDa 이하로 한외여과 하였다. 한외여과 된펩타이드는 ion exchange chromatography와 normal-phase HPLC를 사용하여 칼슘 결합 펩타이드를 분리하였고, 분리된 각 분획의 칼슘결합력을 측정하였다. 그 결과 가장 높은 칼슘 결합력을 보인 분획을 칼슘 chelation을 위한 소재로 얻었고, 따라서 본 연구 결과 얻어진 보리 단백질 가수분해물은 칼슘보충제로의 사용이 가능하다고 판단된다.

• Journal of Periodontal and Implant Science
• /
• 제42권5호
• /
• pp.166-172
• /
• 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
• /
• 제55권4호
• /
• pp.263-266
• /
• 2012

면실박으로부터 단백질을 추출한 후 단백질 가수분해효소인 Flavourzyme으로 가수분해를 실시하여 면실박 단백질 가수분해물을 얻었고, 가수분해 정도는 trinitrobenzenesulfonic acid 방법과 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis를 통해 측정하였다. 면실박 단백질 가수분해물은 한외여과에 의하여 3 kDa 이하로 cut-off하였고, Q-Sepharos fast flow, Sephadex G-15, reversed-phase high performance liquid chromatography를 이용하여 Fe, Ca-binding 펩타이드를 분리하였다. 그 결과 철분과 칼슘 결합력이 가장 높은 분획 51을 얻을 수 있었고, 이렇게 얻어진 Fe, Ca-binding 펩타이드는 향후 기능성 식품 소재로써 활용될 수 있다고 판단된다.

• 대한수의학회지
• /
• 제41권2호
• /
• pp.139-146
• /
• 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.