• 한국축산식품학회지
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• 제40권3호
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• pp.415-425
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• 2020

It is believed that two main proteolytic systems are involved in the tenderization of meat: the cathepsins and the calpains. Many researchers consider the calpain system to be the major contributor to meat tenderness during post-mortem storage. However, the role and activity of cathepsins during post-mortem storage or low temperature meat processing is unclear, particularly for the tough meat cuts like brisket. Thus, the study was designed to investigate the effects of cold (refrigerated and frozen) storage and sous vide processing on the activities of cathepsin B, H, and L in beef brisket. There were no significant changes in pH and cathepsin H activity throughout the 18 d of storage at both temperatures. However, an increase in cathepsin B activity was observed during the first 4 d at both storage temperatures, but subsequently the activity remained unchanged. Cathepsins B and L were found to be more heat stable at sous vide temperatures (50℃ for 24 h, 55℃ for 5 h and at 60℃ and 70℃ for 1 h) compared to cathepsin H. Cathepsin B+L activity was found to increase after sous vide cooking at 50℃ for 1 h but decreased to about 47% relative to the uncooked control after 24 h of cooking. These results suggest that cathepsins B and L may contribute to the improved meat tenderness usually seen in sous vide cooked brisket meat.

• 한국미생물·생명공학회지
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• 제27권6호
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• pp.458-465
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• 1999

Streptomyces luteogriseus KT-10 isolated from Korean farm soil produced a strong cathepsin B inhibitor. Optimal conditions for the cathepsin B inhibitor production by s. luteogriseus KT-10 were evaluated. The cathepsin B inhibitor was produced with maximal yield in the cultural condition of pH 7.0 and $25^{\circ}C$ for 4 days. Optimal medium for the cathepsin B inhibitor production was determined to be a medium containing 20g, peptone 3g, yeast extract 1g, K2HPO4 0.5g, MgSO4.7H2O 0.5g, NaNO3 0.5g, NaCl 0.5g per l. The cathepsin B inhibitor produced by S. luteogriseus KT-10 could also inhibit the other proteinases such as trypsin, papain, and cathepsin D.

• Journal of Oral Medicine and Pain
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• 제33권1호
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• pp.35-40
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• 2008

고농도 세포외 포도당이 치주인대세포의 integrin 발현과 cathepsin-B 및 -L의 발현에 미치는 영향을 살펴보기 위하여 사람 치주인대로부터 일차배양을 통해 얻은 치주인대세포를 1,000 mg/L 농도의 포도당이 포함된 배양액(대조군)과 4,500mg/L 농도의 포도당이 포함된 배양액(실험군)으로 나누어 24시간과 48시간 배양하였다. 그 후, RT-PCR을 통하여 integrin, cathepsin-B 및 -L의 발현을 평가하여 다음과 같은 결론을 얻었다. 1. 대조군에 비해 실험군에서 ${\alpha}5$ intergrin 발현이 증가하였다. 2. Cathepsin-B는 24시간 배양 실험군에서 발현이 증가하였으나, 48시간 배양후에는 발현이 감소하였다. 3. Cathepsin-L은 24시간과 48시간 배양군 모두에서 대조군에 비해 실험군에서 발현이 감소하였다. 이상의 결과로 보아, 고농도 포도당 조건은 치주인대세포의 integrin 발현을 증가시키며, 이는 세포활성에 영향을 미쳐 치주조직의 재생을 지연시킬 것으로 추측된다. 또한, 이 과정은 cathepsin 발현의 감소로 인해 촉진될 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제7권5호
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• pp.333-340
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• 1997

An actinomycetes, KT-10 isolated from ginseng field in Kyongpook, Korea was selected based on its ability to produce a lysosomal cathepsin B inhibitor. The inhibitor purified from the culture supernatant of the isolate KT-10 showed strong inhibitory effects against cathepsin B as well as against papain when the activities were measured using synthetic substrate, ${\alpha}$-N-benzyloxycarbonyl-L-Iysine p-nitrophenyl ester (CLN) or ${\alpha}$-N-benzoyl-D,L-arginine 2-naphthylamide (BANA). The isolate KT-10 was identified as a species of Streptomyces based on its morphological characteristics and chemotaxonomic data. The TAXON program of Ward was used to identify Streptomyces sp. KT-10 as a strain of Streptomyces luteogriseus belong to cluster 18 of the genus Streptomyces with a Willcox probability 0.999388. The cathepsin B inhibitor was presumed to a novel material composed of a polyhydroxylamine.

• 한국미생물·생명공학회지
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• 제29권2호
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• pp.84-89
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• 2001

포유동물조직의 세포내에 존재하는 lysosomal cysteine계 cathepsin B 효소는 암 발병과 전이, 류머티스 관절염, 염증 및 노인성치매 등의 여러 질병에 관여하고 있다. 이 cathepsin B를 저해하는 저해물질이 Streptomyces luteogriseus KT-10에 의해 생산되어 분리되었다. S. luteogriseus KT-10이 생산하는 cathepsin B 저해물질은 열에 안정하며 산, 알칼리에도 안정한 물질로 구성되어 있다. Cathepsin B 저해물질은 DEAE-Sephadex A-25, Sephadex G-15, silica gel, Sephadex LH-20의 column chromatography 과정을 거친 후 분취용 HPLC를 이용 분취한 후 백색분말의 cathepsin B 저해물질을 정제하였다. 이 정제한 저해물질은 UV 상의 전 범위에서 특이한 검출부위를 확인할 수 없었으며, $H_2$O, methanol, ethanol, n-butanol에는 녹지만 비극성인 chloroform, n-hexane, benzene 등에는 불용성이었다. 또한 ninhydrine, $H_2$$SO_4$, iodine에 양성 반응이지만 Ehrlichs, Pauly, Sakaguchi, phthalic acid, DNS, aniline 등의 단백반응과 당 반응에는 음성 반응으로 나타났다. IR 기기에서는 OH 기와 CH 기의 peak를 확인하였으며 $^1$H NMR 기기로 OH peak와 NH peak 또한 확인할 수 있었다. $^{13}$ C NMR spectrum은 4개의 탄소 peak를 가진 물질로 확인된 분자량이 207 dalton인 저해물질이다. 원소 분석기를 이용한 저해물질 분석 결과 분자식이 $C_4$$H_{11}$ $O_4$$N_{6}$로 나타났다. 이 cathepsin B 저해물질의 저해양식은 competitive 저해로 작용함을 확인할 수 있었다.

• 한국식품저장유통학회지
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• 제12권3호
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• pp.275-281
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• 2005

우슬이 민간요법으로 관절염 치료에 우수한 한약재로 알려진 바 우슬 추출물이 cathepsin B 에 대한 저해력을 검토하기 위하여 각종 용매로 우슬 추출물을 순차 분획하고 분획 추출물에 대하여 cathepsin B에 대한 저해활성을 검토하였으며 우슬의 지표물질인 20-hydroxy ecdysone이 추출분획에 검출되는지 TLC 및 HPLC를 이용하여 분석하였으며 우슬의 지표물질이 cathepsin B에 대한 저해활성여부를 확인한바 다음과 같은 결과를 얻었다. 우슬 뿌리에 대해 methanol/water(4:1, v/v) 추출물을 ethyl acetate, chloroform, chloroform/methanol(3:1, v/v), methanol의 각 용매로 분획한 결과, 분획 F4(methanol 분획)에서 수율이 가장 높아 $8.27\%$를 나타내었다. 각 분획별에 따라 cathepsin B에 대한 저해활성을 측정한 결과 F4분획물에서 가장 저해활성이 높았으며 F1분획에서도 높은 저해활성을 나타내었다. F4분획물 중 우슬의 지표물질인 20-hydroxy ecdysone이 검출되었으며 우슬에 함유되어있는 함량은 $0.33\%$이었다. 20-hydroxy ecdysone에 대하여 cathepsin B저해활성을 보기 위하여 cathepsin B저해활성제로 알려진 leupeptin과 활성을 비교해본 결과 기질을 BANA로 사용하였을 경우 leupeptin은 저해율이 $92\%$인데 비하여 20- hydroxy ecdysone은 $88\%$이었으며 F4분획물은 $97\%$를 나타내었고 기질을 CLN으로 사용하였을 경우 leupeptin은 저해율이 $62\%$인데 비하여 20-hydroxy ecdysone은 $36\%$이었으며 F4추출물은 $67\%$를 나타내었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제23권1호
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• pp.129-135
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• 2011

Cathepsins are well-characterized proteases that are ubiquitously expressed in lysosomes. Previous work revealed that $Bombyx$ $mori$ cathepsins B and D are expressed in the fat body and undergo decomposition during larval-pupal metamorphosis. Quantitative RT-PCR was performed to detect cathepsin gene expression at the transcription level when challenged by $B.$ $mori$ nuclear polyhedrosis virus (BmNPV), temperature and hormones (20-hydroxyecdysone (20E) and juvenile hormone analogue (JHA)). mRNAs encoding cathepsins B and D were significantly enhanced after the larvae were infected with BmNPV, and the peak of the induction appeared at 1 day before spinning. This attenuated the inducing effect on cathepsin expression caused by infection. Temperature shock induced cathepsin expression at the later stage of the $5^{th}$ instar, and transcription levels varied with development stage and temperature. Cathepsin B and D mRNA expression in the fat body were significantly induced by JHA at the day before spinning, and with 20E, the expression reached a peak at the last day of the $5^{th}$ instar. Cathepsin B and D mRNA expression exhibited detectable changes post-treatment, without significant differences between or among the hormone concentrations.

• Reproductive and Developmental Biology
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• 제37권4호
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• pp.175-183
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• 2013

Cathepsin B is abundantly expressed peptidase of the papain family in the lysosomes, and closely related to the cell degradation system such as apoptosis, necrosis and autophagy. Abnormal degradation of organelles often occurs due to release of cathepsin B into the cytoplasm. Many studies have been reported that relationship between cathepsin B and intracellular mechanisms in various cell types, but porcine embryos has not yet been reported. Therefore, this study evaluated the effect of cathepsin B inhibitor (E-64) on preimplantation developmental competence and quality of porcine embryos focusing on apoptosis and oxidative stress. The expression of cathepsin B mRNA in porcine embryos was gradually decreased in inverse proportion to E-64 concentration by using real-time RT-PCR. When putative zygotes were cultured with E-64 for 24 h, the rates of early cleavage and blastocyst development were decreased by increasing E-64 concentration. However, the rate of blastocyst development in $5{\mu}M$ treated group was similar to the control. On the other hand, both the index of apoptotic and reactive oxygen species (ROS) of blastocysts were significantly decreased in the $5{\mu}M$ E-64 treated group compared with control. We also examined the mRNA expression levels of apoptosis related genes in the blastocysts derived from $5{\mu}M$ E-64 treated and non-treated groups. Expression of the pro-apoptotic Bax gene was shown to be decreased in the E-64 treated blastocyst group, whereas expression of the anti-apoptotic Bcl-xL gene was increased. Taken together, these results suggest that proper inhibition of cathepsin B at early development stage embryos improves the quality of blastocysts, which may be related to not only the apoptosis reduction but also the oxidative stress reduction in porcine embryos.

• 수산해양교육연구
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• 제28권4호
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• pp.903-912
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• 2016

Hagfish which belongs to the chordate contact cyclostomata, is important phylogenetic relationship between vertebrate and invertebrate. Cathepsins of the cysteine protease family have traditionally been thought to play a major role in intracellular protein degradation and turnover in lysosomes. In this study, Catepsin L was cloned from Inshore hagfish (Eptatretus burgeri), the cDNA encoding ORF of the Eptatretus burgeri Cathepsin L (EbCtL) is 978 bp. The cDNA encoding proEbCtL was expressed in Escherichia coli strain BL21(DE3) using the pGEX-4T-1 expression vector system. The recombinant proEbCtL protein was overexpressed as a approximately 55 kDa fusion protein. The overproduced soluble GST-fusion protein was then applied to glutathione-Sepharose 4B column chromatography; the sample harboring the fusion protein evidenced a high degree of purity when analyzed via SDS-PAGE and Western blot analysis. Its activity was quantied by cleaving the synthetic peptide Z-FR-AMC, Z-LLE-AMC, and Suc-AAF-AMC, and the optimal pH for the protease activity was 8, 9.5, and 9, respectively.

• Bulletin of the Korean Chemical Society
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• 제32권9호
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• pp.3459-3464
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• 2011

In order to extend the scaffold of non-peptidic calpain inhibitor, we have designed and synthesized 14 chalcone derivatives categorized into two groups based on their structures. Compounds 7 ($IC_{50}=16.67{\pm}0.42{\mu}M$) and 8 ($IC_{50}=16.92{\pm}0.14{\mu}M$) in group A were most selective ${\mu}$-calpain inhibitor over cathepsins B and L. On the other hand, compound 14 possessing furan ring exhibited inhibitory activities for ${\mu}$-calpain ($IC_{50}=15.39{\pm}1.34{\mu}M$) as well as cathepsin B ($IC_{50}=20.59{\pm}1.35{\mu}M$). The results discovered implicated that chalcone analogues possessing proper size and functional groups can be a potential lead core for selective non-peptidic ${\mu}$-calpain inhibitor. Furthermore, dual inhibitors for ${\mu}$-calpain and cathepsin B can also be developed from chalcones by elaborate structure manipulation.