• Fisheries and Aquatic Sciences
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• v.15 no.3
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• pp.183-190
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• 2012

The purpose of this study was the purification and characterization of an angiotensin I converting enzyme (ACE) inhibitory peptide purified from enzymatic hydrolysates of rainbow trout Oncorhynchus mykiss muscle. After removal of lipid, the approximate composition analysis of the rainbow trout revealed 24.4%, 1.7%, and 68.3% for protein, lipid, and moisture, respectively. Among six hydrolysates, the peptic hydrolysate exhibited the highest ACE inhibitory activity. We attempted to purify ACE inhibitory peptides from peptic hydrolysate using high performance liquid chromatography on an ODS column. The $IC_{50}$ value of purified ACE inhibitory peptide was $63.9{\mu}M$. The amino acid sequence of the peptide was identified as Lys-Val-Asn-Gly-Pro-Ala-Met-Ser-Pro-Asn-Ala-Asn, with a molecular weight of 1,220 Da, and the Lineweaver-Burk plots suggested that they act as a competitive inhibitor against ACE. Our study suggested that novel ACE inhibitory peptides purified from rainbow trout muscle protein may be beneficial as anti-hypertension compounds in functional foods.

• The Journal of Korean Medicine
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• v.31 no.5
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• pp.64-81
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• 2010

Objectives: The object of this study was to observe the suppressive effects of Yijintang-gamibang (YJGMB), Yijintang being traditionally used in the Korean Medicine for treating various digestive diseases, on the rat reflux esophagitis (RE) as compared with omeprazole, a well-known proton pump inhibitor. Methods: Three different dosages of YJGMB, 50, 100 and 200 mg/kg, were orally pretreated once a day for 28 days before pylorus and forestomach ligation. Seven groups of 8 rats each were used in the study. Six hrs after pylorus and forestomach ligation, changes to the stomach and esophagus lesion areas, gastric volumes, acid and pepsin outputs, invasive lesion percentages, fundic mucosa, esophageal submucosa and total thicknesses were measured by histomorphometry. The results were compared with omeprazole 10 and 30 mg/kg treated groups in which the effects on RE were already confirmed. Results: As results of pylorus and forestomach ligation, marked increases of esophageal and gastric mucosa lesion areas, gastric volumes, acid outputs, pepsin outputs were observed with histopathological changes of RE, such as hemorrhages, ulcerative lesions and edematous changes on the esophageal and fundic mucosa. However, these pylorus and forestomach ligation induced RE were dose-dependently inhibited by treatment of 50, 100 and 200 mg/kg of YJGMB. YJGMB 50 mg/kg showed similar suppressive effects as 30 mg/kg of omeprazole, but more favorable effects were observed as compared with omeprazole 10 mg/kg. Conclusion: The results suggest that YJGMB showed favorable suppressive effects on the RE induced by pylorus and forestomach ligation. It is therefore expected that YJGMB will show favorable effects on RE as corresponds to the suggestion of traditional Korean medicine. However, more detailed mechanism studies should be conducted in future with the screening of the biological active chemical compounds in herbs.

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.124-129
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• 2016

The extraction and purification of the anti-hyperglycemic α-glucosidase inhibitor from an edible mushroom, Pleurotus cornucopiae, were investigated. The inhibitor was maximally extracted when the P. cornucopiae fruiting body was treated with distilled water at 30℃ for 12 h. Purification was achieved using Sephadex G-100 and G-50 filtration chromatography, pepsin hydrolysis, and reverse-phase HPLC. The compound’s solid yield and inhibitory activity were 12.2% and 9.10 mg/ml of IC₅₀, respectively. The purified inhibitor contained two hexapeptides with Thr-Ile-Ala-Phe-Ile-Asp (A) and Tyr-Tyr-Ala-Ile-Gly-Asp (B) sequences and molecular weights of 678.79 Da (A) and 643.7 Da (B). The purified inhibitor showed a mixed inhibition pattern to α-glucosidase and a dose-dependent anti-hyperglycemic effect in a streptozotocininduced diabetic Sprague-Dawley rat model, exhibited by decreased blood glucose levels at doses of 50 and 300 mg/kg.

• Korean Journal of Food Science and Technology
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• v.26 no.1
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• pp.81-87
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• 1994

Two different trypsin inhibitors, TRTI-1 and TRTI-2, were purified to near homogenity from Trichosanthes kirilowii root, by $0{\sim}90%$ saturated ammonium sulfate salting out, DEAE-Sephacel ion exchange chromatography, Sephadex G-50 gel filtration chromatography and trypsin-affinity chromatography. The molecular weight of TRTI-1 and TRTI-2 were estimated to be about 5,000 Da and 24,000 Da, respectively, by gel filtration and must be monomer and homodimer since they contain 4,000 Da and 10,000 Da each on SDS-polyacrylamide gel electrophoresis. TRTI-1 was stable after heating for at least 2 hr at $100^{\circ}C$ but TRTI-2 was completely inactivated after heating for 10 min at $90^{\circ}C$. When Bz-dl-Arg-pNA was used as a substrate of TPCK-treated trypsin, half-maximal inhibitions of TRTI-1 and TRTI-2 were observed at $0.8\;{\mu}M$ and 6\;${\mu}M$, repectively. Both TRTI-1 and TRTI-2 inhibited the hydrolysis of trypsin competitively and Km values were $0.97\;{\mu}M$ and $0.63\;{\mu}M$, respectively. Both TRTI-1 and TRTI-2 specifically inhibited trypsin but they did not inhibit other proteases tested, chymotrypsin, papain, elastase, collagenase, thermolysin, Nagarase, pepsin, and thrombin.

• The Journal of Internal Korean Medicine
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• v.31 no.1
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• pp.128-141
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• 2010

Purpose : The object of this study was to observe the suppressive effects of Yijintang-gamibang (YJGMB), Yijintang with Atractylodis Rhizoma, Massa Medicata Fermentata, Hordei Fructus Germiniatus, and Coptidis Rhizoma. YJGMB has been traditionally used in Korean medicine for treating various digestive diseases. We tested it on the rat reflux esophagitis (RE) induced by pylorus and forestomach ligation in rats as compared with omeprazole, a well-known proton pump inhibitor. Method : Three different dosages of YJGMB 200, 100 and 50mg/kg, were orally pretreated once a day for 28 days before pylorus and forestomach ligation. Seven groups, each of 8 rats per group were used in the study. Six hours after pylorus and forestomach ligation, changes of the stomach and esophagus lesion areas, gastric volumes, acid and pepsin outputs, invasive lesion percentages, fundic mucosa and total thicknesses were measured as histomorphometry. The results were compared with omeprazole, antioxidant and proton pump inhibitor, and 30 and 10mg/kg treated groups in which the effects on RE were already confirmed. Results : As results of pylorus and forestomach ligation, marked increases of esophageal and gastric mucosa lesion areas, gastric volumes, acid outputs, pepsin outputs were observed with histopathological changes of RE, such as hemorrhages, ulcerative lesions and edematous changes on the fundic mucosa. However, these pylorus and forestomach ligation-induced RE were dose-dependently inhibited by treatment of 200, 100 and 50mg/kg of YJGMB. YJGMB 200mg/kg showed similar protective effects as compared with 30mg/kg of omeprazole in the present study, and more favorable effects were observed in 50mg/kg of YJGMB treated rats as compared with omeprazole 10mg/kg in the present study. Conclusion : The results obtained in this study suggest that YJGMB has favorable protective effects on the RE induced by pylorus and forestomach ligation. Therefore, it is expected that YJGMB will also show favorable effects on RE corresponding well to the suggestion of traditional Korean medicine. However, more detailed mechanism studies should be conducted in future with the screening of the biological active chemical compounds in herbs.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.5
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• pp.769-777
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• 2004

A Helicobacter pylori urease inhibitor from Rubus coreanus Miquel has been isolated and partially characterized for aiming to Prevent H. pylori growth and decrease harmful accumulation of ammonia in human gastric mucosa. We screened urease inhibitory activities in 519 extracts library prepared by solvent extraction from 173 kinds of edible plants, medicinal herbs, herbs and seaweeds using a colorimetric urease assay system. As results of primary and secondary screening, 70% acetone extract of Rubus coreanus Miquel was selected as potent candidate, showing about 24% inhibitory activity. The acetone extract was sequentially partitioned into RCE/RCWI and RCB/RCW2 layers with ethyl acetate and butanol. The major active component in RCW2, water layer from butanol fractionation was revealed to be peptidic or proteinous substance by inhibitory activity determination after pronase digestion and periodate oxidation. RCW2-IIIc a was isolated by sequential column chromatography on DEAE-Toyopearl 650C, Butrl-Toyopearl 650M and Sephadex LH-20. The isolated urease inhibitor RCW2-IIIc $\alpha$, was highly pure proteinous substance with molecular weight of 13kDa by high-performance gel permeation liquid chromatography. RCW2-IIIc$\alpha$ has about 5 times higher inhibitory activity than 70％ acetone extract, showing high stability against heat treatment and peptic digestion.

• Journal of Microbiology and Biotechnology
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• v.16 no.12
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• pp.1904-1911
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• 2006

The cell-free extracts of 250 yeasts were screened for their in vitro anti-angiogenic activity, to develop a new cancer metastasis inhibitor. Saccharomyces cerevisiae K-7 was selected as the producer of the anti-angiogenic agent, because it had the highest anti-angiogenic activity. The anti-angiogenic agent was produced maximally from hydrolysates of Saccharomyces cerevisiae K-7, when the yeast was cultured in yeast extract-peptone-dextrose medium at 30$^{\circ}C$ for 24 h, and cell-free extracts were than digested with pepsin for 4 h at 37$^{\circ}C$. The anti-angiogenic agent was further purified by ultrafiltration, Sephadex G-25 gel permeation chromatography and reverse-phase HPLC, and the anti-angiogenic activity of the final purified preparation was 72.7% at 10 $\mu$M/egg. The purified anti-angiogenic agent was found to originate from the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) molecule of Saccharomyces cerevisiae K-7, and its peptide sequence was Val-Ser-Trp-Tyr-Asp-Asn-Glu-Tyr-Gly-Tyr-Ser-Thr-Arg-Val-Val-Asp. In the MTT assay, the shape of the HT-l 080 cell was clearly changed to a circular type at 0.2 mM purified anti-angiogenic agent. This result indicated that the growth of the HT-I080 cell was significantly inhibited at 0.2 mM of the purified anti-angiogenic agent. The MMP activity of the treated HT-l080 cells was not affected, evidenced by the gelatin zymography, indicating that the anti-angiogenic mechanism of the purified anti-angiogenic agent is not mediated through MMP activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.3
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• pp.367-375
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• 1998

The physicochemical properties of the $\alpha$-amylase inhibitors from black bean and naked barley is Korea were investigated. Preincubation time for maximum inhibition was 30min and no activity change was seen after that time. Optimum pH of the $\alpha$-amylase inhibitors from the black bean and naked barley was pH 7.0 and the inhibitory activities were stable in the range of pH 6.0~8.0 in both phosphate and Tris-HCI buffer solutions. Both inhibitors maintained more than 50% of activity after incubation for 17 min at 7$0^{\circ}C$. The inhibitors from the black bean and naked barley maintained more than 50% of activities after treatment for 40 min and 30 min with pepsin, and 30 min and 50 min with trypsin, respectively. Both inhibitors functioned via a noncompetitive mechanism and were active against porcine pancreatic and human salivary $\alpha$-amylases. The activities of both inhibitors were linear for the ionic stength ranging from 0 to 0.9. The addition of 70 mM maltose to the reaction mixture caused a maximum increase in the relative activities of both inhibitors, but it did not affect the dissociation of the EI complex. The activities of both inhibitors were significantly enhanced by adding 1mM of K+ or Mg2+.

• Korean Journal of Pharmacognosy
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• v.35 no.1 s.136
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• pp.52-61
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• 2004

Inhibitory activities of 264 methanol extracts, which were prepared from different parts of 210 kinds of wild and medicinal plants, against human thrombin were evaluated. Based on the anti-coagulation activity determined by thrombin time and activated partial thromboplastin time, the 14 extracts were screened. The fibrinolytic activity, heat stability and inhibition of other proteolytic digestive enzymes, such as pepsin, papain, trypsin and chymotrypsin, of the 14 extracts were further determined, and Ginko biloba (herba), Ephedra sinica (radix), Reynoutria elliptica (herba), Amomum tsao-ko Crevost (fructus), and Magnolia officinalis Rehd. et Wils (bark) were finally selected as possible plant sources for anti-thrombosis agent. These results suggested that medicinal and wild plants could be the potential source of thrombin inhibitor.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.1
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• pp.40-50
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• 1990

The purpose of this paper is to develop a colorant from krill, Euphausia superba, process wastes for use in food products. Carotenoproteins were extracted from preboiled krill processing offal(PKPO) and raw frozen krill processing offal(RKPO) with the aid of proteolytic enzymes. The long-term stability of the astaxanthin associated with the carotenoprotein by the addition of pretense inhibitor and antioxidant to the product were also investigated. Total astaxanthin contents of PKPO and RKPO were $35.1mg\%,\;22.1mg\%$ and those in carotenoproteins were $98.6mg\%,\;61.9mg\%$, respectively. The chitin contents of PKPO and RKPO were $6.9\%,\;4.5\%$, however, those of carotenoproteins were not determined. When $0.5\%$ trypsin was added to the extraction medium containing 0.5M $Na_3EDTA$ at $4^{\circ}C,\;74\%$ of astaxanthin and $83\%$ of the protein of PKPO were recovered as carotenoprotein in 24hrs. The amino acid profile in carotenoprotein was mainly composed of glutamic acid, methionine, aspartic acid and isoleurine. Their contents amounted to about 40% of the total amino acids, followed by alanine, phenylalanine, Iysine, leucine, threonine and tyrosine in that order, with a small amount of cysteine and tryptophan. The levels of essential amino acids were high as much as $38.3\%\~43.6\%$ of the total amino acids. The maximum observance of the carotenoid fraction from krill processing offal and from carotenoprotein was 469nm in petroleum ether. The separated components of carotenoprotein by TLC had Rfs $0.20\~0.23\;0.56\~0.60$ and $0.88\~0.91$. The carotenoids were comprised of astaxanthin, astaxanthin monoester and asthaxanthin diester in $25\~30\%\;,35\~40\%$and $40\~45\%$, respectively. The loss of carotenoids in the carotenoprotein can be prevented by the addition of pro-tease inhibitor(trasylol) and antioxidant(BHT) below $4^{\circ}C$.