• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1318-1323
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• 2004

This study describes the purification and characterization of a novel antihypertensive angiotensin 1­converting enzyme (ACE) inhibitory peptide from Saccharomyces cerevisiae. Maximal production of the ACE inhibitor from Saccharomyces cerevisiae was obtained from 24 h of cultivation at $30^{\circ}C$ and its ACE inhibitory activity was increased by about 1.5 times after treatment of the cell-free extract with pepsin. After the purification of ACE inhibitory peptides with ultrafiltration, Sephadex G-25 column chromatography, and reverse-phase HPLC, an active fraction with an $IC_{50}$ of 0.07 mg and $3.5\%$ yield was obtained. The purified peptide was a novel decapeptide, showing very low similarity to other ACE inhibitory peptide sequences, and its amino acid sequence was Tyr-Asp-Gly-Gly-Val-Phe-Arg-Val-Tyr-Thr. The purified inhibitor competitively inhibited ACE and also showed a clear antihypertensive effect in spontaneously hypertensive rats (SHR) at a dosage of 1 mg/kg body weight.

• 한국식품과학회지
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• 제31권3호
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• pp.848-854
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• 1999

전통된장으로부터 안지오텐신전환효소(angiotensin converting enzyme; ACE)를 저해하는 물질을 추출하여 그 구조를 밝혀냈다. ACE 저해물질을 열수추출한 다음 gel permeation chromatography (GPC)를 통하여 ACE 저해작용이 큰 두 개의 큰 획분을 받았다. 앞획분은 90%와 70%의 ACE 저해효과를 나타내었으나 단일물질로 분리되지 않아 계속하여 역상 HPLC를 통하여 순수 분리를 하였다. 그러나 앞획분은 순수분리되지 않아 결국 2차원 전기영동/TLC를 통하여 분리한 결과 분자량이 759.63인 아미노기를 갖고 있는 비펩타이드 물질임이 밝혀졌다. 뒷획분은 다른 조건의 HPLC(reverse column과 $NH_2$, column)를 이용하여 순수분리에 성공하였다. 이중 ACE 저해효과가 큰 물질은 분자량 271.33인 dipeptide인 arginine-proline임을 밝혀냈다. 이물질의 ACE $IC_{50}$은 $92\;{\mu}M$이었다. 본 연구 결과는 대부분 ACE 저해물질이 3개 내지 7개 등의 긴 펩타이드임을 감안할 때, 짧은 dipeptide로 ACE 저해펩타이드가 한국의 전통된장에서 생산할 수 있음을 보여주고 있다.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.339-342
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• 2012

This study describes the characterization of a new angiotensin I-converting enzyme (ACE) inhibitory peptide from a Korean traditional rice wine. After purification of the ACE inhibitor peptides with ultrafiltration, Sephadex G-25 column chromatography, and successively $C_{18}$ and SCX solid-phase extraction, reverse-phase HPLC, and size exculsion chromatography, two types of the purified ACE inhibitors with $IC_{50}$ values of 0.34 mg/ml and 1.23 mg/ml were finally obtained. The two purified ACE inhibitors (F-1 and F-2) were found to have two kinds of novel oligopeptides, showing very little similarity to other ACE inhibitory peptide sequences. The amino acid sequences of the two purified oligopeptides were found to be Gln-Phe-Tyr-Ala-Val (F-1) and Ala-Gly-Pro-Val-Leu-Leu (F-2), and their molecular masses were estimated to be 468.7 Da (F-1) and 357.7 Da (F-2), respectively. They all showed a clear antihypertensive effect on spontaneously hypertensive rats at a dosage of 500 mg/kg.

• Applied Biological Chemistry
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• 제41권4호
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• pp.270-272
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• 1998

Angiotensin converting enzyme (ACE) inhibitor was isolated from beef bone extract hydrolysate. After hydrolysis of beef bone extract with a commercial protease, ACE inhibitory peptide was purified by using ultrafiltration, gel permeation chromatography, and reverse-phase high pressure liquid chromatography. The purified ACE inhibitor was a pentapeptide, Gly-Pro-X-Gly-Pro.

• 한국식품영양과학회지
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• 제32권7호
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• pp.1120-1125
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• 2003

숙성기간(1, 3 및 3년)이 다른 멸치액젓으로부터 추출한 peptide는 정제과정에서 숙성 1년의 멸치액젓은 3개, 숙성 3년의 멸치액젓은 4개, 숙성 5년의 멸치액젓은 5개의 peak를 각각 나타내었다. 숙성기간이 길수록 멸치액젓의 Peptide는 저분자화 되었으며, gel chromatography상의 후반부에 나타났다. 숙성기간이 다른 멸치액젓으로부터 추출한 peptide들의 생리활성 (항산화, 항암, ACE 저해활성)은 숙성기간이 길수록 증가하였다. 특히 항산화 및 항암활성은 $IC_{50}$/이 각각 34 및 44 $\mu\textrm{g}$/mL인 3년 숙성 peak 3이 제일 높았으나, ACE 저해활성 은 $IC_{50}$/이 32 $\mu\textrm{g}$/mL인 1년 숙성 peak 3이 제일 높았다. 천연 숙성 멸치액젓의 저분자 peptide는 뛰어난 생리활성을 갖고 있음이 밝혀졌으나 구조분석 등의 추가 연구가 필요하다고 본다.

• 한국식품과학회지
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• 제27권2호
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• pp.230-234
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• 1995

식품 유래의 생리활성 peptide를 분리할 목적으로 전통발효식품인 된장으로부터 혈압강하기능을 가지는 ACE(angiotensin converting enzyme)저해활성 peptide를 분획하였다. 시판 된장의 동결건조분말을 냉수로 추출했을 때 총질소의 회수율은 추출시간 30분에 73.3%를 보였다. 용매추출액에서 고분자 polypeptide를 제거하고 저분자 peptide만을 얻기 위해 PM-10 membrane(Amicon)을 이용하여 3시간 동안 한외여과한 결과, 질소성분의 회수율은 80.8%, 염의 회수율은 99.2%에 달했고 투과액의 ACE $IC_{50}$은 $41.8{\mu}g/ml$이었다. 한외여과 투과액을 reverse phase prep-HPLC로 분획하여 7개의 획분을 얻었으며, 그 중 F5분획물의 ACE저해활성이 $IC_{50}=6.8{\mu}g/ml$로 비활성이 가장 높았다. 염은 F1분획물에서 모두 회수되었다. F5분획물을 ion exchange prep-HPLC로 다시 분획하여 얻은 5개의 모든 획분에서 높은 비활성을 보였다($IC_{50}=2.5{\sim}8.3{\mu}g/ml$). 그 중 F53분획물의 비활성이 가장 높았으며($IC_{50}=2.5{\mu}g/ml$), F53의 구성아미노산 분석 결과 histidine의 함량이 특징적으로 높았다.

• Journal of Animal Science and Technology
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• 제47권1호
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• pp.83-90
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• 2005

ACE-inhibitory peptides derived from goat's whey hydrolyzed by various proteolytic enzymes were separated and purified for antihypertension materials. The highest ACE-inhibitory activity of goat's whey hydrolysates was 85.5 % by pepsin for 72 hrs. Also the highest ACE-inhibitory activity of goat's whey hydrolysates was F-4 by pepsin for 72 hrs by Sephadex G-25 gel chromatograms. F-4e and F-4ed from F-4 by RP-HPLC to first and second purification were the highest in ACE-inhibitory activity, respectively. The most abundant amino acid was leucine(I 8.54 %) in F-4ed of ACE-inhibitory peptides after second purification. Amino acid sequence of F-4ed of ACE-inhibitory peptides showed Leu-Lys-Asp-Tyr-Gly-GlyVal- Ser-Leu and Leu-Gly-Asp-Gly-Ala-Gly- Asp-Val-Ala-Phe. $IC_{50}$ calibrated in peptic hydrolysates(72 hrs), F-4, F-4e and F-4ed from goat's whey hydrolysates by pepsin for 72 hrs were 33.93, 28.75, 11.74 and 1.09 mg/ml, respectively. From the results of this experiment, goat's whey hydrolysate by pepsin was shown to have ACE-inhibitory activity.

• 한국수산과학회지
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• 제32권4호
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• pp.427-431
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• 1999

해조자원의 기능 특성을 밝혀 그 이용도를 증진시키기 위한 연구의 일환으로 일반 해조류의 단백질 함량 및 물추출물과 해조 가수분해물의 ACE 저해효과를 조사한 결과는 다음과 같다. 1 단백질 함량은 김이 $39.6\%$로 가장 높았으며, 파래 $22.1\%$, 미역 $21.1\%$, 우뭇가사리 $18.3\%$, 청각 $15.7\%$, 톳 $12.4\%$, 다시마 $8.2\%$의 순으로 나타났다. 2. 해조 물추출물의 ACE 저해효과는 $50^{\circ}C,\;70^{\circ}C,\;98^{\circ}C$의 세 조건에서 각각 추출한 결과 $70^{\circ}C$에서 추출시 시료 모두에서 가장 높게 나타났으며, ACE 저해율이 가장 높은 $70^{\circ}C$를 기준으로 비교했을 때 우뭇가사리 $10.9\%$, 김 $9.3\%$, 파래 $8.9\%$, 미역 $8.2\%$, 톳 $7.5\%$, 다시마 $7.1\%$, 청각 $7.0\%$로 나타났다. 3. maxazyme과 papain에 의한 해조 가수분해물의 ACE 저해효과는 김이 파래, 미역, 우뭇가사리에 비해 월등히 높았고 pep-tide-nitrogen함량 역시 매우 높았다. 김의 경우 가수분해 8시간에서 ACE 저해효과가 maxazyme $31.3\%$, papain $27.9\%$로 가장 높았고 peptide-nitrogen함량도 이 시간에서 가장 높게 나타났다. 파래의 ACE 저해효과 역시 8시간에서 maxazyme $9.6\%$, papain $8.5\%$로 다른 시간대에 비해 대체로 높았으며 peptide-nitrogen함량은 16시간까지 완만히 증가하였다.

• Fisheries and Aquatic Sciences
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• 제14권3호
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• pp.174-178
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• 2011

This study aimed to determine the degree of hydrolysis and angiotensin-I-converting enzyme (ACE)-inhibitory activity of Giant Jellyfish Nemopilema nomurai (jellyfish) hydrolysates. The degree of hydrolysis using six proteolytic enzymes (Alcalase, Flavozyme, Neutrase, papain, Protamex, and trypsin) ranged from 13.1-36.8% and the inhibitory activities from 20.46-79.58%. Using papain hydrolysate, we newly isolated and characterized ACE-inhibitory peptides with a molecular weight of 3,000-5,000 Da that originated from jellyfish collagen. The purified peptide (FII-b) was predicted to be produced from an alpha-2 fragment of the type IV collagen of jellyfish. The N-terminal sequence of FII-b was Asp-Pro-Gly-Leu-Glu-Gly-Ala-His-Gly- and showed 87% identity to the collagen type IV alpha-2 fragment of Rattus norvegicus and a predicted protein from Nematostella vectensis, indicating that the ACE-inhibitory peptide originated from the collagen hydrolysate and had an $IC_{50}$ value of 3.8 ${\mu}g$/mL. The primary structure of the fragment is now being studied; this peptide represents an interesting new type of ACE inhibitor and will provide knowledge of the potential applications of jellyfish components as therapies for hypertension.

• Journal of Microbiology and Biotechnology
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• 제16권5호
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• pp.757-763
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• 2006

Angiotensin I-converting enzyme (ACE) inhibitors have generally been very useful to remedy or prevent hypertension. This study describes the extraction and characterization of an ACE inhibitor from the fruiting body of Pholiota adiposa ASI 24012, which can be used as an antihypertensive drug. The maximal ACE inhibitory activity $(IC_{50};0.25mg)$ was obtained when the fruiting body of Pholiota adiposa ASI 24012 was extracted with distilled water at $30^{\circ}C$ for 12 h. After the purification of ACE inhibitor with ultrafiltration, Sephadex G-25 column chromatography, and reverse-phase HPLC, an active fraction with an $IC_{50}$ of 0.044 mg was obtained. The purified ACE inhibitory peptide was a novel pentapeptide, showing very little similarity to other ACE inhibitory peptide sequences. The molecular mass of the purified ACE inhibitor was estimated to be 414 daltons with a sequence of Gly-Glu-Gly-Gly-Pro, and showed a clear antihypertensive effect on spontaneously hypertensive rats (SHR) at a dosage of 1 mg/kg.