• Food Science of Animal Resources
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• v.40 no.3
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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.

• Microbiology and Biotechnology Letters
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• v.27 no.6
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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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• v.33 no.1
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• pp.35-40
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• 2008

This experiment was designed to clarify the effect of high concentrations of extracellular glucose on the periodontal ligament cells. The cells were incubated for 24 and 48 hours with ${\alpha}-MEM$ including 1,000 mg/L(control group) and 4,500 mg/L(experimental group) of glucose. Then, the expressions of ${\alpha}5$ integrin, cathepsin-B and -L were examined using RT-PCR method. The results were as follows: 1. ${\alpha}5$ integrin expression was increased after incubation in high glucose media. 2. Cathepsin-B expression was increased after the 24-H incubation with high glucose media, but was decreased after the 48-H incubation. 3. The expression of cathepsin-L was decreased by the high glucose media. Theses results suggest that extracellular glucose at high concentration may be attributed to delayed wound healing in diabetes patients through increase in ${\alpha}5$ integrin and decrease in cathepsins, which lead to accumulation of extracellular matrices and adhesion molecules.

• Journal of Microbiology and Biotechnology
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• v.7 no.5
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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.

• Microbiology and Biotechnology Letters
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• v.29 no.2
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• pp.84-89
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• 2001

Isolation and Characterization of Cathepsin B inhibitor Produced by Streptomyces luteogriseus KT-IO. Han, Kil~Hwan and Sang~Dal Kim*. Department of Applied Microbiology, Yeungnam Universit}/t Kyongsan 712749, Korea - The cathepsin B inhibitor produced by Streptomyces luteogriseus KT-IO was very stable in heat, acidic and alkaline conditions. The cathepsin B inhibitor was isolated from the extracted fraction of culture broth with butanol, methanol and chloroform subsequently, the inhibitor was purified with following several column chromatography sLlch as DEAE-Sephadex A-25, Sephadex G-15, silica gel 60, Sephadex LH-20, and preparative HPLC. The cathepsin B inhibitor showed positively to detective reaction of ninhydrine, 5% H2S04, iodine, but negatively to the reaction of Ehrlich's reagent, DNS, aniline. The molecular formular of cathepsin B inhibitor was elucidated by JR, lH and 13C-NMR, FAB mass and elemental analyzer. Consequently, it was identified as C4HlI04N6. The cathepsin B inhibitor had the mode of competitive inhibition with the reaction of cathepsin B.

• Food Science and Preservation
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• v.12 no.3
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• pp.275-281
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• 2005

This study was carried out to investigate the cathepsin B inhibition effect by Achyranthes japonica N. root extract in vitro. The methanol/$H_{2}O$(4:1, v/v) extract was fractionated by ethyl acetate(F1), chloroform(F2), chloroform/methanol(3:1, v/v)(F3) and methanol(F4). The yield of F4 in Achyranthes japonica N. root was $8.27\%$. As an index material of Achyranthes japonica N. root, 20-hydroxy ecdysone was detected by TLC, and HPLC and it's content was $0.33\%$. Three isolates(F1, F3, F4) showed the cathepsin B inhibition activity, and F4 showed the highest inhibition activity among them. In the inhibition activity on cathepsin B, leupeptin, 20-hydroxy ecdysone and F4(at the same concentration of 20-hydroxy ecdysone.) were 92, 88 and $97\%$ on BANA($N{\alpha}$-benzoyl-DL-arginine ${\beta}$-naphthylamide) substrate, and 62, 36 and $67\%$ on CLN($N{\alpha}$-CBZ(carbobenzlyoxy)-L-lysine p-nitrophenyl ester HCI) substrate, respectively.

• International Journal of Industrial Entomology and Biomaterials
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• v.23 no.1
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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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• v.37 no.4
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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.

• Journal of Fisheries and Marine Sciences Education
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• v.28 no.4
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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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• v.32 no.9
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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.