• 한국수산과학회지
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• 제42권5호
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• pp.417-425
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• 2009

The potential utility of fish scales to the functional food industry has been investigated due to its antioxidant and antihypertensive characteristics. In this study, we report on the reactive oxygen species (ROS) scavenging and angiotensin I converting enzyme (ACE) inhibitory activities of gelatin hydrolysates processed from Branchiostegus japonicus scales, which are also high in protein content (about 46.1%). We prepared the enzymatic gelatin hydrolysates with four proteases (${\alpha}$-chymotrypsin, Alcalase, Neutrase and trypsin) from B. japonicus scale gelatin, which was prepared according to different reaction times, substrate/enzyme ratios and substrate concentrations. The enzymatic hydrolytic degrees of the gelatin increased time-dependently up to 6 hrs, while the Alcalase gelatin hydrolysates showed the highest hydrolysis degrees compared to the others. Furthermore, gelatin hydrolysates of Neutrase and ${\alpha}$-chymotrypsin showed the highest DPPH radical and $H_2O_2$ scavenging activities ($IC_{50}$ value; 9.18 mg/mL and 9.74 mg/mL), respectively. However, the activities were not significant (P<0.05). We also observed that the four gelatin hydrolysates significantly increased ACE inhibitory activities from approximately 20% to 60% (P<0.05), Among them, the Alcalase gelatin hydrolysates showed the higher ACE inhibitory activity ($IC_{50}$ value; 0.73 mg/mL) compared to the others. These results suggest that the enzymatic gelatin hydrolysates prepared from B. japonicus scales may possess a potentially useful function as an ACE inhibitory agent. As such, the utility of B. japonicus scales should be given due consideration for application in the functional food industry.

• Journal of Microbiology and Biotechnology
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• 제26권3호
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• pp.461-468
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• 2016

In recent years, various naturally occurring defence peptides such as plectasin have attracted considerable research interest because they could serve as alternatives to antibiotics. However, the production of plectasin from natural microorganisms is still not commercially feasible because of its low expression levels and weak stability. A tandemly arrayed plectasin gene (1,002 bp) from Pseudoplectania nigrella was generated using the isoschizomer construction method, and was inserted into the pPICZαA vector and expressed in Pichia pastoris. The selected P. pastoris strain yielded 143 μg/ml recombinant plectasin (Ple) under the control of the methanol-inducible alcohol oxidase 1 (AOX1) promoter. Ple was estimated by SDS-PAGE to be 41 kDa. In vitro studies have shown that Ple efficiently inhibited the growth of several gram-positive bacteria such as Streptococcus suis and Staphylococcus aureus. S. suis is the most sensitive bacterial species to Ple, with a minimum inhibitory concentration (MIC) of 4 μg/ml. Importantly, Ple exhibited resistance to pepsin but it was quite sensitive to trypsin and maintained antimicrobial activity over a wide pH range (pH 2.0 to 10.0). P. pastoris offers an attractive system for the cost-effective production of Ple. The antimicrobial activity of Ple suggested that it could be a potential alternative to antibiotics against S. suis and S. aureus infections.

• 대한소아치과학회지
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• 제30권4호
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• pp.626-636
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• 2003

치태형성을 억제하는 유산균 K1270을 김치로부터 분리한 후 배양적, 생화학적 특징 및 16S rDNA염기서열 분석에 의해 Pediococcus pentosaceus K1270으로 동정하였다. K1270 균주는 5% 자당이 함유된 배지에서 Streptococcus mutans Ingbritt에 의한 인공치태 형성을 대조군에 비해 94.6 % 억제하였으며, 비수용성 글루캔의 합성을 89.6 % 억제하였다. S. mutans Ingbritt의 증식은 대조군에 비해 100배 정도 억제하였다. K1270균주는 TMB와 peroxidase가 첨가된 MRS 한천배지에서 과산화수소를 생산하였으며, 인공치태 형성에 대한 K1270균주의 억제 효과는 catalase 첨가에 의해 일부 감소되었다. K1270 균주의 배양 상청액을 10% 자당이 함유된 $2{\times}M17$ broth에 동량 가한 경우 인공치태 형성 및 Streptococcus mutans Ingbritt의 증식이 억제되었으며, 이 억제효과는 catalase첨가에 의해 일부 감소되었고, 열처리, 또는 trypsin 처치에 의해 완전히 소실되었다. 따라서, 본 연구에서 분리, 동정된 P. pentosaceus K1270은 과산화 수소와 bacteriocin 유사물질을 분비하여 S. mutans Ingbritt의 증식을 억제함으로써 인공치태 형성을 억제하는 것으로 사료된다.

• 한국연초학회지
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• 제13권2호
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• pp.27-33
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• 1991

Current studies indicated that emphysema in smokers might be due, in part, to the local suppression of G, -protease inhibitor(u, -Pl) in lung by reactive oxygen species in cigarette smoke or smoke-activated lung neutrophiles. In the present works, we examined the possibility that a measure which inactivated $\alpha$l-Pl by cigarette smoke could be an alternative method to evaluate the cigarette quality, In order to determine the inactivation of $\alpha$1, -Pl, trypsin inhibitory capacity(TIC) was assayed. A rapid loss of $\alpha$1, -Pl activity occurred when $\alpha$1-Pl solutions was exposed the gas phase or total particulate matter(TPM) obtained from various brands. The inactivation of $\alpha$1-Pl by gas phase was dependent upon the number of puffs and the age of the smoke. However, that by TPM was rather decreased since 2 puffs and also showed no more change over 24hrs after exposing. Inactivation of $\alpha$1-Pl determined by our suggested method(5 puffs, 24hours of aging after exposing) using various commercial cigarettes exhibited that high tar brands has inactivated it more strongly than low tar cigarettes. But the ability of some brands to inactivate $\alpha$1-Pl does not correlate with the content of tar or nicotine. These results so여esc that the degree of $\alpha$1-Pl inactivation by cigarette smoke may be a useful index for the evaluation of cigarette quality and that it should be also contribute to the manufacture of less hazardous cigarettes.

• 대한한방내과학회지
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• 제32권2호
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• pp.188-202
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• 2011

Objectives : Oxidative stress seems to play a major role in mechanisms by which ethanol causes liver injury. Previous studies have shown that treatment with Chungganhaeju-tang (Qingganjiejiu-tang, CGHJT) has protective effects on alcoholic liver disease. The aim of this study was to investigate the effects of Chungganhaeju-tang on oxidative stress. Materials and Methods : In vitro, we evaluated the inhibitory activities of CGHJT on DPPH (1,1-diphenyl-2-picryl-hydrazyl), xanthine oxidase, trypsin, and hyaluronidase, and measured cell viability, and proliferation. In the cell culture model, we measured the activities of superoxide dismutase (SOD), and catalase (CAT) after CGHJT treatment in C34 and E47 cell lines, HepG2 cells transfected with/without the cytochrome P450 2E1 (CYP2E1) gene. In vivo, we measured malondialdehyde levels in the liver tissue and alcohol concentration in the blood. Results : CGHJT showed significant free radical scavenging activity against DPPH and xanthine oxidase in the in vitro study, and increased cell viability, proliferation, and activities of superoxide dismutase, catalase in C34 and in E47 cell lines. CGHJT reduced malondialdehyde levels and blood alcohol concentration in vivo, as well. Conclusions : This study suggests that CGHJT has antioxidant effects on oxidative stress by reducing lipid peroxidation and inhibiting the ethanol induced suppression of antioxidant enzyme activities.

• Asian-Australasian Journal of Animal Sciences
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• 제22권11호
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• pp.1587-1593
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• 2009

Duck egg white (DEW) hydrolysates were prepared by five enzymes (papain, trypsin, chymotrypsin, alcalase, and flavourzyme) and their antioxidant activities investigated in this study. DEW hydrolyzed with papain (DEWHP) had the highest peptide content among the five enzymatic treatments. Besides, the peptide content of DEWHP increased when the enzyme to substrate ratio (E/S ratio) increased. It was suggested that higher E/S ratio contributed to elevate the degree of hydrolysis of DEW effectively. Similar results were also obtained by Tricine-SDS-PAGE. In addition, SDS-PAGE patterns indicated papain was the only one amongst all enzymes with the ability to hydrolyze DEW. In antioxidant properties, DEWHP showed more than 70% of inhibitory activity on linoleic acid peroxidation and superoxide anion scavenging. Moreover, the $Fe^{2+}$ chelating effect of DEWHP was greater than 90%, while no significant difference was observed in DPPH radical scavenging and reducing ability. The results of peptide contents, antioxidant activities and electrophoresis suggested that the higher the peptide content, the stronger the antioxidant activities in DEWHP.

• Preventive Nutrition and Food Science
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• 제7권4호
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• pp.347-352
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• 2002

The present study was undertaken to evaluate the anti-rheumatoidal arthritis effect of the R. verniciflua heartwood extract, its EtOAc fraction, and its primary flavonoids, sulfuretin and fustin. All test samples showed variably significant inhibitory effects on hind paw edema and trypsin inhibitor activity induced by Freund's complete adjuvant reagent (FCA reagent), and on vascular permeability caused by acetic acid. Treatment with 10 mg/kg (i.p.) sulfuretin for seven days inhibited edema formation by 54.2$\pm$3.0%. Test samples, especially sulfuretin, shifted the values of biochemical parameters such as serum-cholesterol, serum-triglyceride and serum-total protein toward the normal and restored the numbers of leucocytes and platelets. These results suggest that the heartwood of R. verniciflua reduces immunological injuries caused by FCA reagent provides evidence that suluretin is an active anti-rheumatoid arthritis agent.

• Archives of Pharmacal Research
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• 제26권12호
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• pp.1024-1028
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• 2003

A prolyl endopeptidase inhibitor was isolated from the ethyl acetate soluble fraction of Phyllanthus ussurensis. The active compound was identified as an ellagitannin, corilagin. It was shown to non-competitively inhibit prolyl endopeptidase (PEP) with the $IC_{50}$ value of $1.17 \times $10^{-6}\mu$M. The Ki value was $6.70 \times 10^{-7}$ M. Corilagin was less inhibitory to other serine proteases such as chymotrypsin, trypsin, and elastase, indicating that it was relatively a specific inhibitor of PEP. Corilagin also effectively inhibited reactive oxygen species such as hydroxide and superoxide anion radical, hydrogen peroxide, and DPPH. Especially, corilagin showed potent scavel1ging activity on the superoxide anion radical in the ESR method ($IC_{50} =3.79 \times 10^{-6}$M) as well as xanthine oxidase system.

• Food Science and Biotechnology
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• 제15권4호
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• pp.617-624
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• 2006

Various plant phenolics were assessed for (${\beta}$-secretase (BACE1) inhibitory activity in order to screen for anti-dementia agents. Among 39 phenolics, eight compounds, 1,2,3-trigalloyl glucopyranoside, acetonyl geraniin, euphorscopin, furosine, helioscopinin A, helioscopinin B, jolkinin, and rugosin E exhibited strong inhibition of BACE1 with $IC_{50}$ values of $5.87{\times}10^{-8}-54.93{\times}10^{-6}\;M$. Among them, rugosin E was the most potent ($IC_{50}$ $5.87{\times}10^{-8}\;M$). The active compounds were shown to be non-competitive inhibitors by Dixon plot. All the phenolic BACE1 inhibitors except furosin also suppressed prolyl endopeptidase (PEP) activity. However, these phenolic compounds caused less inhibition of ${\alpha}$-secretase (tumor necrosis factor a converting enzyme; TACE) and no significant inhibition of other serine proteases such as trypsin, chymotrypsin, and elastase was seen, demonstrating that they are relatively specific to both BACE1 and PEP. No significant structure-activity relationships were found.

• Journal of Microbiology and Biotechnology
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• 제21권6호
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• pp.617-626
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• 2011

Transglutaminase from Streptomyces mobaraensis is an enzyme of unknown function that cross-links proteins to high molecular weight aggregates. Previously, we characterized two intrinsic transglutaminase substrates with inactivating activities against subtilisin and dispase. This report now describes a novel substrate that inhibits papain, bromelain, and trypsin. Papain was the most sensitive protease; thus, the protein was designated Streptomyces papain inhibitor (SPI). To avoid transglutaminase-mediated glutamine deamidation during culture, SPI was produced by Streptomyces mobaraensis at various growth temperatures. The best results were achieved by culturing for 30-50 h at $42^{\circ}C$, which yielded high SPI concentrations and negligibly small amounts of mature transglutaminase. Transglutaminasespecific biotinylation displayed largely unmodified glutamine and lysine residues. In contrast, purified SPI from the $28^{\circ}C$ culture lost the potential to be cross-linked, but exhibited higher inhibitory activity as indicated by a significantly lower $K_i$ (60 nM vs. 140 nM). Despite similarities in molecular mass (12 kDa) and high thermostability, SPI exhibits clear differences in comparison with all members of the wellknown family of Streptomyces subtilisin inhibitors. The neutral protein (pI of 7.3) shares sequence homology with a putative protein from Streptomyces lavendulae, whose conformation is most likely stabilized by two disulfide bridges. However, cysteine residues are not localized in the typical regions of subtilisin inhibitors. SPI and the formerly characterized dispase-inactivating substrate are unique proteins of distinct Streptomycetes such as Streptomyces mobaraensis. Along with the subtilisin inhibitory protein, they could play a crucial role in the defense of vulnerable protein layers that are solidified by transglutaminase.