• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.700-707
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• 2010

A novel antioxidative peptide (APVPH I, antioxidative peptides from venison protein hydrolysates I) was purified from venison by enzymatic hydrolysis, column chromatography of DEAE-Sephacel, and high-performance liquid chromatography. The molecular mass of the purified peptide was found to be 9,853 Da and the amino acid sequences of the purified peptide was Met-Gln-Ile-Phe-Val-Lys-Thr-Leu-Thr-Gly. The purpose of this study was to evaluate the effects of APVPH I against $H_2O_2$-induced neuronal cells damage in PC-12 cells. Antioxidative enzyme levels in cultured neuronal cells were increased in the presence of the peptide. In addition, APVPH I inhibited productions of nitric oxide (NO), reactive oxygen species (ROS), malondialdehyde (MDA), and cell death against $H_2O_2$-induced neuronal cell damage in PC-12 cells. It was presumed to be APVPH I involved in regulating the apoptosis-related gene expression in the cell environment. The present results indicate that APVPH I substantially contributes to antioxidative properties in neuronal cells.

• Bulletin of the Korean Chemical Society
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• 제26권12호
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• pp.1911-1920
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• 2005

Effective artificial metalloproteases have been designed by using cross-linked polystyrene as the backbone. Artificial active sites comprising Cu(II) complexes as the catalytic site and other metal centers or organic functionalities as binding sites were synthesized. The activity of Cu(II) centers for peptide hydrolysis was greatly enhanced on attachment to polystyrene. By placing binding sites in proximity to the catalytic centers, the ability to hydrolyze a variety of protein substrates at selected cleavage sites was improved. Thus far, the most advanced immobile artificial proteases have been obtained by attaching the aldehyde group in proximity to the Cu(II) complex of cyclen.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2012년도 제42회 동계 정기 학술대회 초록집
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• pp.557-557
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• 2012

Silicatein-${\alpha}$, the enzyme extracted from silica spicules in glass sponges, has been studied extensively in the way of chemistry from 1999, in which the pioneering work by Morse, D. E. - the discovery of the enzymatic hydrolysis in Silicatein-${\alpha}$ - was published. Since its reaction conditions are physiologically favored, synthesis of various materials, such as gallium oxide, zirconium oxide, and silicon oxide, was achieved without any hazardous wastes. Although some groups synthesized oxide films and particles, they have not achieved yet controlled morphogenesis in the reaction conditions mentioned above. With the knowledge of catalytic triad involved in hydrolysis of silicone alkoxide and oligomerization of silicic acid, we designed the novel peptide amphiphiles to not only form self-assembled structure, but also display similar activities to silicatein-${\alpha}$. Designed templates were able to self-assemble into left-handed helices for the peptide amphiphiles with L-form amino acid, catalyzing polycondensation of silicic acids onto the surface of them. It led to the formation of silica helices with 30-50 nm diameters. These results were characterized by various techniques, including SEM, TEM, and STEM. Given the situation that nano-bio-technology, the bio-applicable technology in nanometer scale, has been attracting considerable attention; this result could be applied to the latest applications in biotechnology, such as biosensors, lab-on-a-chip, biocompatible nanodevices.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.899-902
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• 2005

Peptide assimilation by Helicobacter pylori was investigated using L-phenylalanyl-3-thia-phenylalanine (PSP) as a detector peptide; the release of thiophenol upon enzymatic hydrolysis of PSP was spectrophotometrically detected with the aid of 5,5'-dithiobis[2-nitrobenzoic acid] (DTNB). By adding PSP to whole-cell suspension, thiophenol was produced progressively, resembling that found in Esherichia coli or Staphylococcus aureus. Interestingly, the rate of thiophenol production by H pylori in particular was markedly reduced when cells were pretreated with trypsin, indicating surface exhibition of peptidase. According to the competitive spectrophotometry using alanyl-peptides, H pylori did not appear to assimilate PSP through the peptide transport system. No discernible PSP assimilation could be ascertained in H pylori cells, unless provided with some additives necessary for peptidase activity, such as $Ni^{2+}\;or\;Mg^{2+}$ and an appropriate concentration of potassium or ammonium salts. These observations strongly suggest that, regardless of a presumptive peptide transport system, peptide assimilation of H. plori appears to be highly dependent upon milieu conditions, due to unique peptidase exhibition on the cell surface.

• 한국축산식품학회지
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• 제34권2호
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• pp.151-157
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• 2014

This study investigated the effects of three proteases (trypsin, pepsin and chymotrypsin) on the hydrolysis efficiency of porcine placenta and the molecular weight (Mw) distributions of the placental hydrolysates. Because placenta was made up of insoluble collagen, the placenta was gelatinized by applying thermal treatment at $90^{\circ}C$ for 1 h and used as the sample. The placental hydrolyzing activities of the enzymes at varying concentrations and incubation times were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel permeation chromatography (GPC). Based on the SDS-PAGE, the best placental hydrolysis efficiency was observed in trypsin treatments where all peptide bands disappeared after 1 h of incubation as compared to 6 h of chymotrypsin. Pepsin hardly hydrolyzed the placenta as compared to the other two enzymes. The Mw distribution revealed that the trypsin produced placental peptides with Mw of 106 and 500 Da. Peptides produced by chymotrypsin exhibited broad ranges of Mw distribution (1-20 kDa), while the pepsin treatment showed Mw greater than 7 kDa. For comparisons of pre-treatments, the subcritical water processing (37.5 MPa and $200^{\circ}C$) of raw placenta improved the efficiency of tryptic digestions to a greater level than that of a preheating treatment ($90^{\circ}C$ for 1 h). Consequently, subcritical water processing followed by enzymatic digestions has the potential of an advanced collagen hydrolysis technique.

• 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.

• Fisheries and Aquatic Sciences
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• 제21권5호
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• pp.13.1-13.8
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• 2018

Amyloid plaque, also called senile plaque, the product of aggregation of ${\beta}$-amyloid peptides ($A{\beta}$), is observed in brains of the patients with Alzheimer's disease (AD) and is one of the key factors in etiology of the disease. In this study, hydrolysates obtained from the sea hare (Aplysia kurodai) were investigated for ${\beta}$-secretase inhibitory peptide. The sea hare's muscle protein was hydrolyzed using six enzymes in a batch reactor. Trypsin hydrolysate had highest ${\beta}$-secretase inhibitory activity compared to the other hydrolysates. ${\beta}$-secretase inhibitory peptide was separated using Sephadex G-25 column chromatography and high-performance liquid chromatography on a C18 column. ${\beta}$-secretase inhibitory peptide was identified as eight amino acid residues of Val-Ala-Ala-Leu-Met-Leu-Phe-Asn by N-terminal amino acid sequence analysis. $IC_{50}$ value of purified ${\beta}$-secretase inhibitory peptide was $74.25{\mu}M$, and Lineweaver-Burk plots suggested that the peptide purified from sea hare muscle protein acts as a competitive inhibitor against ${\beta}$-secretase. Results of this study suggest that peptides derived from sea hare muscle may be beneficial as anti-dementia compounds in functional foods or as pharmaceuticals.

• 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.

• 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.

• 한국식품영양학회지
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• 제10권4호
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• pp.485-490
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• 1997

옥수수 글루텐을 식물성 효소인 파파인으로 가수분해하여 글루텐 펩타이드를 생산하였다. 생산된 글루펜 펩타이드류는 인이온 존재하에서 인이온과 칼슘이온과의 침전형성을 방지하여 칼슘의 용해성을 증진시켰다. 칼슘이온의 용해도는 8.3mg의 펩타이드 존재시 대조구에 비해 5.2배 증대되었다. 산성 아미노산의 함량이 매우 높은 이들 펩타이드류를 Delta pak 칼럼을 이용하여 분획하였으며 이들 분획 가운데 3번째 분획이 가장 높은 칼슘 용해성을 보였다.