• 한국축산식품학회지
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• 제43권1호
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• pp.184-194
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• 2023

Recently, interest in food-derived bioactive peptides as promising ingredients for the prevention and improvement of hypertension is increasing. The purpose of this study was to determine the structure and antihypertensive effect of an antioxidant peptide purified from velvet antler in a previous study and evaluate its potential as a various bioactive peptide. Molecular weight (MW) and amino acid sequences of the purified peptide were determined by quadrupole time-of-flight electrospray ionization mass spectroscopy. The angiotensin I-converting enzyme (ACE) inhibition activity of the purified peptide was assessed by enzyme reaction methods and in silico molecular docking analysis to determine the interaction between the purified peptide and ACE. Also, antihypertensive effect of the purified peptide in spontaneously hypertensive rats (SHRs) was investigated. The purified antioxidant peptide was identified to be a pentapeptide Asp-Asn-Arg-Tyr-Tyr with a MW of 730.31 Da. This pentapeptide showed potent inhibition activity against ACE (IC₅₀ value, 3.72 μM). Molecular docking studies revealed a good and stable binding affinity between purified peptide and ACE and indicated that the purified peptide could interact with HOH2570, ARG522, ARG124, GLU143, HIS387, TRP357, and GLU403 residues of ACE. Furthermore, oral administration of the pentapeptide significantly reduced blood pressure in SHRs. The pentapeptide derived from enzymatic hydrolysate of velvet antler is an excellent ACE inhibitor. It might be effectively applied as an animal-based functional food ingredient.

• Fisheries and Aquatic Sciences
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• 제22권10호
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• pp.22.1-22.7
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• 2019

The functional peptides from protein hydrolysates of various fishery sources have been identified such as antioxidant activity. The main intention of this study was purification and characterization of antioxidative peptide from black eelpout muscle. The antioxidative peptides were purified from black eelpout (Lycodes diapterus) muscle using different proteases. Antioxidant activity of black eelpout hydrolysates was evaluated using DPPH radical scavenging activity. Among six hydrolysates, the pepsin hydrolysate had the highest antioxidant activity compared to the other hydrolysates. Therefore, it was further purified and a peptide with seven amino acid residues of DLVKVEA (784 Da) was identified by amino acid sequence analysis. The EC₅₀ value for scavenging DPPH radicals by purified peptide was 688.77 μM. Additionally, the purified peptide exhibited protective effect against DNA damage induces by oxidation in mouse macrophages (RAW 264.7 cells). The results of this study suggest that black eelpout muscle protein hydrolysate could potentially contribute to development of bioactive peptides in basic research.

• 한국수산과학회지
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• 제44권2호
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• pp.118-125
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• 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.

• 한국수산과학회지
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• 제44권5호
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• pp.456-463
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• 2011

We attempted to isolate a collagenase-1 inhibitory peptide from skate Dipturus chilensis skin protein. The protein from skate skin was digested by various enzymes (alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin) to produce a collagenase-1 inhibitory peptide. The collagenase-1 inhibitory activity of the peptides obtained was measured by gelatin digestion assay. Among the six hydrolysates, pepsin hydrolysate exhibited the highest collagenase-1 inhibitory activity. The peptide showing strong collagenase-1 inhibitory activity was purified by Sephadex G-25 gel chromatography and HPLC using an octadecylsilyls (ODS) column. The amino acid sequence of purified collagenase-1 inhibitory peptide was identified to be Asn-Leu-Asp-Val -Leu-Glu-Val-Phe (961 Da) by quadrupole time of flight (Q-TOF) and electrospray ionization mass spectrometry (ESI-MS) mass spectroscopy. The $IC_{50}$ value of purified peptide was 87.0 ${\mu}M$. Moreover, the peptide did not exhibit cytotoxic effects on human dermal fibroblast cell lines.

• 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.

• Preventive Nutrition and Food Science
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• 제8권1호
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• pp.66-69
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• 2003

An angiotensin converting enzyme (ACE) inhibitory peptide was isolated and purified from the hydrolysates of duck meat protein. Duck meat protein was hydrolyzed using trypsin at 37$^{\circ}C$ for 2 hrs. An ACE inhibitory peptide was purified using membrane filtration, anion exchange chromatography, gel permeation chromatography, fast protein liquid chromatography, normal phase HPLC. The purified inhibitory peptide was identified to be a tetrapeptide, Glu-Asp-Leu-Glu having $IC_{50}$/ value of 85.9 $\mu$M.

• 생명과학회지
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• 제27권1호
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• pp.50-56
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• 2017

졸복(Takifugu pardalis)의 아가미로부터 항균성 펩타이드를 정제하였다. 졸복 아가미의 산 추출물은 Sep-Pak C18에 의해 부분적으로 정제되었으며, 60% 메탄올 분획(RM60)은 Bacillus subtilis KCTC 1021에 대해 높은 항균활성을 나타내었다. 이 RM60을 사용하여 이온교환을 포함한 5단계의 연속적인 HPLC로 정제하였다. 정제된 펩타이드의 분자량과 아미노산 서열분석은 MALDI-TOF MS와 에드만 분해법으로 분석하였다. 이 펩타이드의 분자량은 약 1171.6 Da이었으며, 분석된 이 물질의 부분적인 일차구조서열은 다음과 같다; STKEKAPRKQ. 이 물질과 기존에 알려진 항균성 펩타이드들과 유사성을 조사한 결과, 정제된 물질은 histone H3 계열의 N-말단 부분과 유사하였다. 이러한 결과로부터 정제된 펩타이드는 졸복에서 반응하는 선천성 방어 시스템에서 중요한 역할을 하고 있다고 여겨진다.

• 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.

• 한국식물병리학회지
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• 제14권3호
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• pp.203-208
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• 1998

We isolated and characterized an antifungal peptide from the seeds of Phytolacca americana. Growth inhibition assay with Botrytis cinerea was used to screen inhibitory proteins from 60 different plant species. A 4 kDa antifungal peptide (Pa-AFP) inhibitory to hyphal growth of B. cinerea was found in the seeds of P. americana. The peptide Pa-AFP was purified to homogeneity by chromatographies of Sephadex G-50, DEAE-Sepharose, Sephacryl S-300, and C18 reverse-phase HPLC. Western blot analysis showed that a polyclonal antibody raised against the purified peptide cross-reacted with a 4 kDa protein in seeds but not in root and leaf tissues of P. americana. Pa-AFP inhibited the hyphal growth of Botrytis cinerea, Rihzoctonia solani, Fusarium oxysporum, and Magnaporthe grisea. Pa-AFP exhibited growth inhibition of Saccharomyces cerevisiae strain BWG7a, which was sensitive to osmotin.

• Applied Biological Chemistry
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• 제49권3호
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• pp.196-201
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• 2006

Vitellogenin은 어류의 난황 단백질의 전구체 물질로서 암컷 혈청에서 발견되는 단백질이며, 외부에서 에스트로겐이나 내분비계장애물질에 노출된 경우에는 수컷이나 미성숙한 암컷에서도 이의 합성이 촉진되는 것으로 보고되었다. 따라서 수컷에서 유도되는 vitellogenin은 외인성 에스트로겐 물질에 노출되었음을 암시하는 중요한 지표로 인정되고 있다. Vitellogenin에 대한 항체를 생산하기 위하여 잉어의 vitellogenin 서열 중의 일부분에 대한 peptide를 합성한 후 그 합성 peptide에 대한 항체를 제작하였다. 그리고 $17{\beta}$-estradiol을 주입한 잉어의 혈청에서 DE-52 이온 교환 크로마토그래피를 사용하여 vitellogenin을 정제한 후 이에 대한 항체를 제작하였다. 본 연구에서는 상기의 합성 vitellogenin peptide에 대하여 제작한 다클론 항체와 vitellogenin 단백질에 대한 다클론 항체가 추후 에스트로겐의 지표로 사용될 수 있는 지를 조사하고자 항체의 반응성을 조사하였다. 정제하여 얻은 vitellogenin에 대한 다클론 항체는 Western blotting 시 $17{\beta}$-estradiol을 주입한 잉어의 혈청과 암컷 잉어의 혈청에 있는 vitellogenin과 반응한 반면에 vitellogenin peptide에 대한 다클론 항체는 이들과 반응하지 않았다. 이는 공유결합적으로 많은 변형이 일어난 vitellogenin 단백질은 vitellogenin 합성 펩타이드에 대한 항체로는 검출되지 않음을 나타낸다.