• Korean Journal of Pharmacognosy
• /
• v.44 no.4
• /
• pp.336-343
• /
• 2013

In a continuation of investigation on Cordyceps militaris, thirteen compounds were isolated from the $CH_2Cl_2$ and n-BuOH-soluble fraction of C. militaris. They were identified as twelve diketopiperazines such as cyclo($\small{L}$-Gly-$\small{L}$-Pro) (1), cyclo($\small{L}$-Ala-$\small{L}$-Pro) (2), cyclo($\small{L}$-Ser-$\small{L}$-Pro) (3), cyclo($\small{L}$-Val-$\small{L}$-Pro) (4), cyclo($\small{L}$-Thr-$\small{L}$-Pro) (5), cyclo($\small{L}$-Pro-$\small{L}$-Pro) (6), cyclo($\small{L}$-Thr-$\small{L}$-Leu) (7), cyclo($\small{L}$-Tyr-$\small{L}$-Ala) (8), cyclo($\small{L}$-Phe-$\small{L}$-Ser) (9), cyclo($\small{L}$-Phe-$\small{L}$-Pro) (10), cyclo($\small{L}$-Tyr-$\small{L}$-Pro) (11) and brevianamide F (13), and an amino acid, tryptophan (12). Their structures were identified on the basis of chemical evidences and spectroscopic analysis including 1D-NMR ($^1H$, $^{13}C$), 2D-NMR (HSQC, HMBC) and MS spectral data. Among the isolated compounds, compounds 1, 2, 6-11 are first reported from C. militaris.

• Journal of Dairy Science and Biotechnology
• /
• v.16 no.2
• /
• pp.98-105
• /
• 1998

Various peptides derived from food are among the most potent physiologically active agents known, and include anticancer peptides, angiotensin converting enzyme(ACE) inhibitor exhibiting antihypertension action, opioid peptides, antithrombotic peptides, hypocholesterolemic peptides, immunomodulators, calcium absorption enhancers, and other peptides. Hydrophobic peptides extracted from a Cheddar-type cheese slurry were fractionated by gel chromatography and repeated HPLC. A peptide fraction from HPLC showed high cytotoxicity on the tumor cell lines such as a human colon carcinoma, and comprised of Tyr, Ser, Leu, Gly, and others. Hypocholesterolemic peptides were isolated from peptic hydrolyzates of casein and soy proteins. Macropeptides of 1,000${\sim}$5,000 dalton were effective on reducing the cholesterol level of mouse serum. Peptides showing high Krigbaum hydrophobicity and ANS surface hydrophobicity resulted in high hypocholesterolemic effect and fecal steroid concentrations. Caseinomacropeptides(CMP) were isolated from whey powder and treated with soluble and immobilized trypsin to obtain antithrombotic peptides. One fraction from the CMP hydrolyzed with immobilized trypsin for 24h exhibited high antithrombotic activity with 52.5% inhibition of platelet aggregation. These result suggested that peptides from various milk products could be utilized as a good bioactive agents for developing health functional foods.

• BMB Reports
• /
• v.32 no.6
• /
• pp.579-584
• /
• 1999

Mantis egg fibrolase (MEF-3) was purified from the egg cases of Tenodera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, DEAE Affi-Gel blue gel affinity chromatogragphy, and MONO-Q anion-exchange chromatography. This protease had a molecular weight of 35,600 Da as determined by SDS-polyacrylamide gel electrophoresis under reducing conditions and its isoelectric point was 6.0. The N-terminal amino acids sequence was Ala-Thr-Gln-Asp-Asp-Ala-Pro-Pro-Gly-Leu-Ala-Arg-Arg. This sequence was 80% homologous to the serine protease from Tritirachium album. MEF-3 readily digested the ${\alpha}$-and ${\beta}$-chains of fibrinogen and more slowly the ${\gamma}$-chains. It showed strong proteolytic and fibrinolytic activities. Phenylmethanesulfonyl fluoride and chymostatin inhibited its proteolytic activity, while EDTA, EGTA, cysteine, ${\beta}$-mercaptoethanol, elastinal, tosyl-lysine chloromethylketone, and tosyl-amido-2-phenylethyl chloromethyl ketone did not affect its proteolytic activity. Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEF-3 was benzoyl-Phe-Val-Arg-p-nitroanilide. Based on these experimental results, we speculated that MEF-3 is a serine protease with a strong fibrin(ogen)olytic activity.

• 한국유가공학회:학술대회논문집
• /
• 1998.05a
• /
• pp.22-29
• /
• 1998

Various peptides derived from food are among the most potent physiologically active agents known, and include anticancer peptides, angiotensin converting enzyme(ACE) inhibitor exhibiting antihypertension action, opioid peptides, antithrombotic peptides, hypocholesterolemic peptides, immunomodulators, calcium absorption enhancers, and other peptides. Hydrophobic peptides extracted from a Cheddar-type cheese slurry were fractionated by gel chromatography and repeated HPLC. A peptide fraction from HPLC showed high cytotoxicity on the tumor cell lines such as a human colon carcinoma, and comprised of Tyr, Ser, Leu, Gly, and others. Hypocholesterolemic peptides were isolated from peptic hydrolyzates of casein and soy proteins. Macropeptides of 1,000${\sim}$5,000 dalton were effective on reducing the cholesterol level of mouse serum. Peptides showing high Krigbaum hydrophobicity and ANS surface hydrophobicity resulted in high hypocholesterolemic effect and fecal steroid concentrations. Caseinomacropeptides (CMP) were isolated from whey powder and treated with soluble and immobilized trypsin to obtain antithrombotic peptides. One fraction from the CMP hydrolyzed with immobilized trypsin for 24h exhibited high antithrombotic activity with 52.5% inhibition of platelet aggregation. These results suggested that peptides from various milk products could be utilized as a good bioactive agents for developing health functional foods.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.44 no.2
• /
• pp.118-125
• /
• 2011

In this study, an angiotensin I-converting enzyme (ACE) inhibitor from squid skin was purified and characterized. Squid (Todarodes pacificus) skin protein isolates were hydrolyzed using six commercial proteases: alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin. The peptic hydrolysate had the highest ACE inhibitory activity. The ACE inhibitory peptide was purified using Sephadex G-25 column chromatography and reverse phase high-performance liquid chromatography (HPLC) with a $C_{18}$ column. The purified ACE inhibitory peptide was identified and sequenced, and found to consist of seven amino acid residues: Ser-Ala-Gly-Ser-Leu-Val-Pro (657Da). The $IC_{50}$ value of the purified ACE inhibitory peptide was 766.2 ${\mu}M$, and Lineweaver-Burk plots suggested that the purified peptide acts as a noncompetitive ACE inhibitor. These results suggest that the ACE inhibitory peptide purified from the peptic hydrolysate of squid skin may be of benefit in developing antihypertensive drugs and functional foods.

• Journal of Life Science
• /
• v.10 no.2
• /
• pp.218-227
• /
• 2000

Bone morphogenetic protein 1 (BMP-1) is part of a complex capable of inducing ectopic bone formation in mammals. Studies on TGF-β1 processing and Drosophila dorsal-ventral patterning have focused attention on BMP-1 as important in mediating the biological activity of this bone inducing complex. Herein, the bacterial expression, refolding, purification, and initial characterization of the BMP-1 proteolytic domain (BPD) are described. A semi-quantitative fluorescence-based thin layer chromatography assay was developed to assist in rapidly screening for optimal renaturation conditions. According to a preliminary screen for optimal conditions for the refolding of BPD , a detectable proteolytic activity against a high turnover substrate for astacin, a homologous protease from crayfish was observed. The conditions identified have allowed the expression of sufficient amounts of BPD for the characterization of the protein. Its proteolytic activity exhibits the same cleavage specificity as astacin against seven substrates that were previously synthesized for studying astacin. Furthermore, this activity is inhibited by the metal chelator 1,10-phenanthroline but not by its analogue 1,7-phenanthroline. The collagenase inhibitor Pro-Leu-Gly hydroxamate was found to inhibit both astacin and BPD activity. The results presented in this paper argue that BMP-1 does in fact possess an intrinsic proteolytic activity.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.26 no.6
• /
• pp.1058-1062
• /
• 1997

The objective of this was to investigate the technical feasibility of producing toxicant-free by germination. To this end, rapeseed(Brassica napus L.) was germinated at $25^{\circ}C$ for 120 hours, and the chemical compositions of amino acids and vitamins were determinated in every 24 hours during germination. The results obtained are summarized as follows: Before germination, rapeseed contained 5.4g/16g N of glutamic acid and high percentage of the other amino acids in order of Asp>Leu>His>Pro>Arg>Lys>Gly>Ser>Ala>Val. The amino acids were gradually decreased until 96 hours during germination had tendency to show a slight increase in 120 hours. Vitamin B$_1$, B$_2$and C contents in rapeseed before germination were found to be 0.11, 0.21 and 3.72mg% respectively, and the vitamin E was 423ug/g. The vitamin C greatly increased in 72 hours during germination, while the vitamin B group was drastically decreased in 72 hours. The results obtained by this method clearly demonstrate that germination process is very effective to the removal of toxicants in rapeseed.

• Proceedings of the PSK Conference
• /
• 2002.10a
• /
• pp.330.2-330.2
• /
• 2002

The conformational study on cyclic Ac-Cys-Pro-Xaa-Cys-NHMe (Ac-CPXC-NHMe: X = Ala, Val. Leu. Aib. Gly. His. Phe, Tyr. Asn. and Ser) peptides has been carried out using the ECEPP/3 force field and the hydration shell model in the unhydrated and hydrated states. This work has been undertaken to investigate structural implications of the CPXC sequence as the chain reversal for the initiation of protein folding and as the motif for active site of disulfide oxidoreductases. The backbone conformation DAAA is in common the most feasible for cyclic CPXC peptides in the hydrated state. which has a type 1${\beta}$-turn at the Pro-Xaa sequence. The proline residue and the hydrogen bond between backbones of two cystines appear to play a role in stabilizing this preferred conformation of cycilc CPXC peptides. However. the distributions of backbone conformations and ${\beta}$-turns may indicate that the cyclic CPXC peptide seems to exist as an ensemble of ${\beta}$-turns and coiled conformations. The intirnsic stability of the cyclic CPXC motif itself the active conformation appears to play a role in determining electrochemical properties of disulfide oxidoreductases.

• Journal of Sericultural and Entomological Science
• /
• v.46 no.2
• /
• pp.47-53
• /
• 2004

To reveal the possibility as a source of functional food and to increase the usability of mulberry fruits, amino acid analysis with mulberry fruits was carried out with L-8800 Amino acid auto analyzer. We analyzed 16 amino acid components including 6 essential amino acids. According to the results, the mean content of total amino acid was 4,640.8 mg/100 g DW. 'Sabangso(II)' showed the highest total amino acid content (6,919.3 mg/100 g DW) among the fifty accessions tested, whereas 'Gukkwang' showed the lowest content (3,389.1 mg/100 g DW). The distribution rate of amino acid components decreased in the order of Glu ($20.1\%$) > Gly ($12.7\%$) > Asp ($12.5\%$) > Thr ($8.2\%$) > Ala ($7.9\%$) > Arg ($7.5\%$) > Pro ($6.1\%$) > Ser ($6.1\%$) > Val ($5.7\%$) > Leu ($5.3\%$) > Ile ($2.7\%$) > His ($2.5\%$) > Met ($0.9\%$) > Tyr ($0.8\%$) > Lys ($0.7\%$) > Cys ($0.3\%$). 'Sabangso(II)' showed the highest content in 11 amino acid components, which were Glu (1,293.2 mg/100 g), Ser (390.2 mg/100 g), Thr (564.4 mg/100 g), His (175.3 mg/100 g), Arg (590.4 mg/100 g), Lys (53.5 mg/100 g), Gly (589.4 mg/100 g), Ala (499.3 mg/100 g), Leu (481.9 mg/100 g), Ile (240.9 mg/100 g), and Val (407.6 mg/100 g). Tyr and Cys were the highest in 'Jangsosang' (70.8 mg/100 g, 19.3 mg/100 g, respectively), and Asp, Pro, and Met were the highest in 'Jasan(I)', 'Hwansipchosaeng' and 'Palcheongsipyung(I)' (1,098.8 mg/100 g, 364.4 mg/100 g, 68.8 mg/100 g, respectively).

• Korean Journal of Food Science and Technology
• /
• v.9 no.4
• /
• pp.255-263
• /
• 1977

Fermented squid, Loligo kobiensis, is widely used and occupies an important position in foods of this country. But no study on its taste compounds has been reported. This study was attempted to establish the basic data for evaluating taste compounds of fermented squid. The changes of such compounds during fermentation as free amino acids, nucleotides and their related compounds, TMAO, TMA and betaine were analysed. The sample was prepared with 20% salt content and fermented at a controlled temperature of $15{\pm}3^{\circ}C$. ADP, AMP and inosine tended to degrade rapidly while hypoxanthine increased more than four times as compared with raw sample at 91 day fermentation. In the free amino acid composition of fresh squid, abundant amino acids were proline, taurine, alanine, arginine, serine, glutamic acid, lysine, glycine, leucine and valine in order. Such amino acids like phenylalanine, methionine, tyrosine, isoleucine, and histidine were poor. In squid extract, proline and taurine were dominant holding 40.2% and 32.0% of total free amino acids respectively. The total free amino acid nitrogen in fresh squid was 33.6% of its extract nitrogen. The changes of free amino acid composition in the extract of squid during fermentation was not observed. In the extract of fermented product, abundant amino acids were proline, leucine, lysine, serine, arginine, alanine, valine, isoleucine, phenylalanine, methionine and glycine in order. Glutamic acid and histidine were poor and taurine and tyrosine were trace in content. The increase of total free amino acids during 63 day fermentation reached approximately wore than 1.8 times as compared with that of raw sample and than decreased slowly. The amount of betaine increased more than 1.2 times as compared with that of raw sample during 91 day fermentation. TMA increased while TMAO decreased during fermentation. The amount of TMAO nitrogen in 91 days fermented squid was 402.4mg% on moisture and salt free base. Betaine and TMAO known as sweet compounds were abundant in fermented squid. It is supposed that these compounds could also play a role as important taste compounds of fermented squid. It is concluded that the major taste compounds of fermented squid were amino acids like proline, leucine, serine, lysine, arginine, alanine and betaine. Other compounds such as valine, isoleucine and TMAO and hypoxanthine could also not be excluded as taste supporters in fermented squid.