• Fisheries and Aquatic Sciences
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• 제15권3호
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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.

• 대한한의학회지
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• 제31권5호
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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.

• 한국미생물·생명공학회지
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• 제44권2호
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• pp.124-129
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• 2016

식용버섯인 노랑느타리버섯으로부터 α-glucosidase 저해제의 추출과 정제에 대하여 연구하였다. α-glucosidase 저해제는 노랑느타리버섯 자실체를 증류수로 30℃에서 12시간 처리하였을 때 가장 많이 추출되었다. α-glucosidase 저해제를 sephadex G-100 여과 크로마토그래피와 펩신 가수분해, sephadex G-50 여과 크로마토그래피, 역상 HPLC로 정제한 결과 수율과 α-glucosidase 저해활성은 각각 12.2%와 IC₅₀ 9.10 mg/ml이었다. 정제한 α-glucosidase 저해제는 Thr-Ile-Ala-Phe-Ile-Asp (A)과 Tyr-Tyr-Ala-Ile-Gly-Asp (B)의 서열을 갖고 있는 두개의 헥사펩타이드를 함유하였고 이들의 분자량은 각각 (A)가 678.79 Da, (B)가 643.7 Da이었다. 정제한 α-glucosidase 저해제는 α-glucosidase에 대하여 혼합형 저해양상을 보였고 streptozotocin으로 유도된 당뇨 쥐모델에서 50 mg/kg과 300 mg/kg 투여시 혈당함량을 낮추어 주는, 농도 의존적 혈당상승억제 효과를 보였다.

• 한국식품과학회지
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• 제26권1호
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• pp.81-87
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• 1994

하늘타리박의 뿌리인 괄루근에서 분리 정제한 trypsin inhibitor는 TRTI-1과 TRTI-2 두 종류였으며 gel-filtration과 SDS-PAGE 전기영동에서 얻은 결과로부터 TRTI-1은 $4,000{\sim}5,000\;Da$의 분자량을 가진 monomer이며 TRTI-2는 $20,000{\sim}24,000\;Da$의 분자량을 가친 homodimer임을 알 수 있었다. 온도의 안정성에 있어서는 TRTI-1은 $100^{\circ}C$에서 2시간 이상 안정하였으며, TRTI-2는 $0{\sim}70^{\circ}C$의 온도에서는 안정하였으나 $80^{\circ}C$에서부터는 안정성이 떨어지면서 $100^{\circ}C$에서는 저해능이 완전히 소실되었다. Trypsin 활성에 대해서 TRTI-1, TRTI-2 및 상품화된 soybean BBI의 농도에 따른 저해능을 보면 TRTI-1, TRTI-2 및 soybean BBI의 각각의 농도가 $0.8{\mu}M,\;6{\mu}M,\;4{\mu}M$일 때 1mg/ml의 trypsin을 50% 저해하였으며, trypsin의 반응속도에 대한 TRTI의 저해 실험에서 TRTI-1와 TRTI-2 모두 trypsin의 가수분해를 경쟁적으로 저해하였으며 $K_{m}$값은 저해제가 없는 경우 $0.37\;{\mu}M$에 대해 각각 0.97, $0.63\;{\mu}M$이었다. 여러 protease에 대한 TRTI-1과 TRTI-2의 저해능 실험 결과 elastase, pepsin, Nagarase, papain, thermolysin, chymotrypsin, thrombin 및 collagenase 등은 저해하지 못하였고 trypsin만을 특이하게 저해하였다.

• 대한한방내과학회지
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• 제31권1호
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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.

• 한국식품영양과학회지
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• 제33권5호
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• pp.769-777
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• 2004

위염과 위궤양의 일차적 발병 인자로 알려진 Helicobacter pylori의 생육을 억제하고 urease 산물인 암모니아의 축적에 의한 위점막 손상을 완화시킬 목적으로 복분자로부터 H. pylori urease저해물질을 분리정제하고 소재화와 관련된 일부 성질을 규명하였다. 양념 채소류, 차류, 죽류, 건강채소류등의 식용식물, 약용식물, 허브 및 해조류, 총 173종으로부터 극성도에 따라 계통추출한 수용성인 냉수추출물(Fr, 1), methanol 추출물(Fr.4), 열수추출물(Fr.5) 519점을 대상으로 H. pylori urease 저해활성을 검색하였다. 1차 및 2차 저해활성 검색 결과 복분자의 70% acetone추출물이 약 24%의 가장 높은 저해활성을 나타내었으며, 이 추출물을 ethyl acetate와 butanol을 사용하여 ethyl acetate/DW layer(RCE/RCWl)와 butanol/DW layer(RCB/RCW2)로 순차 분획한 후 활성획분인 RCW2 내의 활성 본체를 확인할 목적으로 periodate oxidation과 pronase digestion을 실시한 결과 펩타이드 또는 단백질성 물질로 판명되었다. 저해활성 물질은 DEAE-Toyopearl 650C, Butyl-Toyopearl 650M 및 Sephadex LH-20순의 column chromatography에 의 해 분리 정제되었다. 분리 urease 저해물질, RCW2-IIIc $\alpha$는 HPLC의 gel permeation chromatography에 의해 비교적 순도 높은 분자량 약 13 kDa의 단일 물질임이 확인되었다. 저해활성물질은 열에 안정성을 보이 는 내열성 단백질임을 알 수 있었고 위내 단백분해효소인 pepsin에도 가수분해 저항성을 나타냄으로써 기능성식품의 소재로 높은 소재화 적성을 보였다.

• Journal of Microbiology and Biotechnology
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• 제16권12호
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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.

• 한국식품영양과학회지
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• 제27권3호
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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+.

• 생약학회지
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• 제35권1호통권136호
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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.

• 한국수산과학회지
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• 제23권1호
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• pp.40-50
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• 1990

크릴가공시 부산물로 얻어지는 잔사를 보다 효율적으로 이용하기 위하여 크릴가공 잔사로부터 carotenoprotein을 추출하여 이를 식품착색료로 이용할 목적으로 호소를 이용한 carotenoprotein의 추출조건 및 품질안정성에 대하여 검토하였다. 크릴가공 잔사중 총 astaxanthin함량은 자숙크릴 잔사 및 생동결 크릴 잔사에서 각각 $35.1mg\%,\;22.1mg\%$ 였으며, $98.6mg\%,\;61.9mg\%$였다. Chitin함량은 크릴가공 잔사인 자숙크릴과 생동결크릴에서 각각 $6.9\%,\;4.5\%$였으나 carotenoprotein에서는 검출되지 않았다. Trypsin이 carotenoprotein추출에 가장 효과적이었으며, papain, pepsin, protease는 거의 효과가 나타나지 않았다. Trypsin 첨가농도 $0.5\%$, 반응온도 $4^{\circ}C$에서 24시간 분해하였을 때 단백질 및 carotenoid의 회수율은 자숙시료가 각각 $83\%,\;74\%$로 가장 좋았다. 생동결크릴의 경우는 자숙크릴에 비해 단백질 및 carotenoid 회수율이 다소 떨어졌다. Carotenoprotein의 아미노산조성중 glutamic acid 와 aspartic acid의 함량이 가장 많았으며 필수 아미노산함량도 전체 아미노산의 $38.3\%\~43.6\%$를 차지하였다. Carotenoprotein중의 carotenoid는 TLC분석에 의해 astaxanthin, astaxanthin monoester, astaxanthin diester로 분리되었으며, 이들의 상대량은 각각 $25\~30\%\;,35\~40\%\;40\~45\%$였다. Carotenoprotein의 안정성은 저온($-20^{\circ}C\;4^{\circ}C$)일수록 안정하였으며, BHT와 protease inhibitor인 trasylol을 동시에 가한 것이 BHT와 trasylol만을 단독으로 가한 것보다 안정성이 좋았다.