• Asian-Australasian Journal of Animal Sciences
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• v.6 no.1
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• pp.19-26
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• 1993

The effect heat treatment (autoclave) on nutritional value of winged bean as compared to soybean has been investigated. The winged bean and soybean were obtained from local cultivar grown in Indonesia. The beans were autoclaved at $120^{\circ}C$ for 15, 30, 45, 60 or 90 minutes, respectively before being ground for chemical analysis. Trypsin inhibitors of winged bean and soybean decreased (p < 0.05) along with decreasing of urease activity as heating time increased from 0 to 90 minutes. Heat treatment significantly (p < 0.05) reduced protein solubility in 0.2% potassium hydroxide of winged bean as well as soybean. In vitro protein digestibility was significantly (p < 0.05) improved by heating treatment (15 to 60 min of autoclaving), however, excessive heating (90 min of autoclaving) decreased the digestibility of winged beans. Excessive heating had adverse effect on lysine, cystine and methionine contents of winged beans. The results of this study suggested that autoclaving at $120^{\circ}C$ within 45 minutes should be adequate to remove protease inhibitors and could improve protein digestibility of winged beans.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.6
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• pp.797-804
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• 1996

Two trypsins were purified from shrimp hepatopancreas through ammonium sulfate fractionation, Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, and Sephacryl S-300 gel chromatography. Both enzymes had a single polypeptide chain with a molecular weight (M.W.) of 32 kDa by sodium dodecylsulfate polyacrylamide gel electrophoresis (SOS-PAGE), although trypsin A and B were estimated to be a molecular weight of 27.2 and 22.8 kDa, respectively, using Sephacryl S-300 gel filtration. Both trypsins had similar amino acid compositions and rich in glycine, valine, alanine, aspartic acid, and glutamic acid, but low in methionine and basic amino acids. Both enzymes were completely inactivated by soybean trypsin inhibitor (SBTI), phenylmethylsulfonyl fluoride (PMSF), tosyl-L-lysine chloromethyl ketone (TLCK), benzamidine, leupeptin, however, not affected by tosyl-L-phenylalanine chloromethyl ketone (TPCK) and pepstatin.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.257-262
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• 1992

Virginiae butanolide (VB) is an autoregulator which triggers virginiamycin production in Strefltomyces virginiae. VB-C binding protein activity was investigated under various additives. The VB-C binding protein was almost fully observed in sotubte fraction (>90%) and the binding activity was optimum at pH 7.0. The VB-C binding activity was increased about 15% in 0.5 M KCI, whereas decreased about 60% in 20 mM $Mo^{6+}$. Chelating reagents (ethylenediarnine tetraacetic acid, ethyleneglycol bis(2-aminoethylether) tetraacetic acid, 8-hydroxyquinoline) and SH protecting reagents (rnercaptoethanol, dithiothreitol, thioglycerol) inhibited the VB-C binding activity about 30~55% and 3~20%, respectively. Serine protease inhibitor (phenyl methane sulfonyl fluoride), nucleotides (guanosine 5'-monophosphate, adenosine 3',5'-cyclic monophosphate), and phosphatases (alkaline, acid phosphatase) increased the VB-C binding activity about 17%, 6~20%, and 4- 13%, respectively.

• Bulletin of the Korean Chemical Society
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• v.30 no.1
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• pp.163-171
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• 2009

Few $\alpha$-Hydroxy-$\beta$-amino acids were synthesized via various nucleophilic addition of the epoxide and followed by stereoselective nucleophilic substitution reaction and eliminative cleavage of the acetal selectively in diacetal compound. One of the synthesized $\alpha$-Hydroxy-$\beta$-amino acid reacted with L-leucine methylester to give corresponding dipeptide in good yields.

• Journal of Life Science
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• v.10 no.2
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• pp.218-227
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• 2000

Bone morphogenetic protein 1 (BMP-1) is part of a complex capable of inducing ectopic bone formation in mammals. Studies on TGF-β1 processing and Drosophila dorsal-ventral patterning have focused attention on BMP-1 as important in mediating the biological activity of this bone inducing complex. Herein, the bacterial expression, refolding, purification, and initial characterization of the BMP-1 proteolytic domain (BPD) are described. A semi-quantitative fluorescence-based thin layer chromatography assay was developed to assist in rapidly screening for optimal renaturation conditions. According to a preliminary screen for optimal conditions for the refolding of BPD , a detectable proteolytic activity against a high turnover substrate for astacin, a homologous protease from crayfish was observed. The conditions identified have allowed the expression of sufficient amounts of BPD for the characterization of the protein. Its proteolytic activity exhibits the same cleavage specificity as astacin against seven substrates that were previously synthesized for studying astacin. Furthermore, this activity is inhibited by the metal chelator 1,10-phenanthroline but not by its analogue 1,7-phenanthroline. The collagenase inhibitor Pro-Leu-Gly hydroxamate was found to inhibit both astacin and BPD activity. The results presented in this paper argue that BMP-1 does in fact possess an intrinsic proteolytic activity.

• BMB Reports
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• v.51 no.12
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• pp.648-653
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• 2018

Serine protease inhibitor Kazal type 1 (SPINK1) plays a role in protecting the pancreas against premature activation of trypsinogen and is involved in cancer progression. SPINK1 promoted LAC cells growth, migration, and invasion. Mechanistically, we found that SPINK1 promoted LAC cells migration and invasion via up-regulating matrix metalloproteinase 12 (MMP12). We observed that SPINK1 expression was only up-regulated in lung adenocarcinoma (LAC) tissues, and was an independent prognostic factor for poor survival. Our results indicate that SPINK1 might be a potential biomarker for LAC that promotes progression by MMP12.

• Animal Bioscience
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• v.35 no.2
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• pp.272-280
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• 2022

Objective: The aim of this study was to evaluate the effect of ageing and feeding strategies on the calpain protease system and meat quality traits in Braford steers. Methods: Thirty Braford steers were employed; 15 animals were supplemented with corn silage during finishing and 15 were kept only on pasture. Meat quality traits and calpain system protein activity were evaluated in longissimus thoracis et lumborum (LTL) steaks aged for 2, 7, 14, and 21 days. Results: Aged meat showed higher pH and calcium content, while Warner Bratzler shear force (WBSF) decreased to day 21. No interaction between ageing and diet was seen for quality traits. Steers finished with corn silage showed higher values of water holding capacity, WBSF and free calcium, and lower values of pH and cooking loss. Calpain and calpastatin activities decreased with ageing. Finishing steers on pasture produced higher values of calpains and lower values of calpastatin activities. The higher values of calpain 1 activity were observed in muscles aged 2 days from pasture finished animals, and the lower activity of the inhibitor in the 21 days aged samples of the same group. Conclusion: These results suggest a diet by ageing interaction in calpains and calpastatin and this interaction impact in Warner Bratzler Shear Force in Braford LTL muscle.

• Journal of Life Science
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• v.26 no.10
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• pp.1189-1195
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• 2016

Vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) are potent angiogenic factors that have been used clinically to induce angiogenesis. To enable migration and invasion, cells must proliferate and secrete proteinases, which degrade the surrounding extracellular matrix. The goal of this study was to investigate the cell proliferation; matrix metalloproteinase-2 (MMP-2), MMP-9, and plasmin secretion; and migration and invasion of glioma-derived U-373-MG cells induced by VEGF and HGF treatment. An additional goal was to test the hypothesis that elevated secretion of MMP-2, MMP-9, and plasmin contributed directly or indirectly to the proliferation, migration, and invasion of U-373-MG cells. Cell proliferation, migration, and invasion and MMP-2, MMP-9, and plasmin secretion were significantly increased in the VEGF and HGF-treated U-373-MG cells. To elucidate the role of the increased secretion of MMP-2, MMP-9, and plasmin in cell proliferation, migration, and invasion of the U-373-MG cells, they were treated with MMPs inhibitor (BB-94) and plasmin inhibitor (α2AP) prior to VEGF or HGF stimulation. The BB-94 and α2AP treatment resulted in a significant reduction in the cell proliferation, migration, and invasion of the U-373-MG cells as compared with the VEGF- and HGF-treated groups. The results indicate that inhibition of MMPs and plasmin reduce the cell proliferation, migration, and invasion of U-373-MG cells.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.1
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• pp.40-50
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• 1990

The purpose of this paper is to develop a colorant from krill, Euphausia superba, process wastes for use in food products. Carotenoproteins were extracted from preboiled krill processing offal(PKPO) and raw frozen krill processing offal(RKPO) with the aid of proteolytic enzymes. The long-term stability of the astaxanthin associated with the carotenoprotein by the addition of pretense inhibitor and antioxidant to the product were also investigated. Total astaxanthin contents of PKPO and RKPO were $35.1mg\%,\;22.1mg\%$ and those in carotenoproteins were $98.6mg\%,\;61.9mg\%$, respectively. The chitin contents of PKPO and RKPO were $6.9\%,\;4.5\%$, however, those of carotenoproteins were not determined. When $0.5\%$ trypsin was added to the extraction medium containing 0.5M $Na_3EDTA$ at $4^{\circ}C,\;74\%$ of astaxanthin and $83\%$ of the protein of PKPO were recovered as carotenoprotein in 24hrs. The amino acid profile in carotenoprotein was mainly composed of glutamic acid, methionine, aspartic acid and isoleurine. Their contents amounted to about 40% of the total amino acids, followed by alanine, phenylalanine, Iysine, leucine, threonine and tyrosine in that order, with a small amount of cysteine and tryptophan. The levels of essential amino acids were high as much as $38.3\%\~43.6\%$ of the total amino acids. The maximum observance of the carotenoid fraction from krill processing offal and from carotenoprotein was 469nm in petroleum ether. The separated components of carotenoprotein by TLC had Rfs $0.20\~0.23\;0.56\~0.60$ and $0.88\~0.91$. The carotenoids were comprised of astaxanthin, astaxanthin monoester and asthaxanthin diester in $25\~30\%\;,35\~40\%$and $40\~45\%$, respectively. The loss of carotenoids in the carotenoprotein can be prevented by the addition of pro-tease inhibitor(trasylol) and antioxidant(BHT) below $4^{\circ}C$.

• The korean journal of orthodontics
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• v.33 no.3 s.98
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• pp.169-183
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• 2003

Tooth movement by orthodontic force effects great tissue changes within the periodontium, especially by shifting the blood flow in the pressure side and resulting in a hypoxic state of low oxygen tension. The aim of this study is to elucidate the possible mechanism of apoptosis in response to hypoxia in MC3T3El osteoblasts, the main cells in bone remodeling during orthodontic tooth movement. MC3T3El osteoblasts under hypoxic conditions ($2\%$ orygen) resulted in apoptosis in a time-dependent manner as estimated by DNA fragmentation assay and nuclear morphology stained with fluorescent dye, Hoechst 33258. Pretreatment with Z-VAD-FMK, a pancaspase inhibitor, or Z-DEVD-CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to hypoxia. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. To confirm what caspases are involved in apoptosis, Western blot analysis was performed using anti-caspase-3 or -6 antibodies. The 10-kDa protein, corresponding to the active products of caspase-3, and the 10-kDa protein of the active protein of caspase-6 were generated in hypoxia-challenged cells in which the processing of the full length form of caspase-3 and -6 was evident. While a time course similar to this caspase-3 and -6 activation was evident, hypoxic stress caused the cleavage of lamin A, which was typical of caspase-6 activity. In addition, the stress elicited the release of cytochrome c into the cytosol during apoptosis. Furthermore, we observed that pre-treatment with SB203580, a selective p38 mitogen activated protein kinase inhibitor, attenuated the hypoxia-induced apoptosis. The addition of SB203S80 suppressed caspase-3 and -6-like protease activity by hypoxia up to $50\%$. In contrast, PD98059 had no effect on the hypoxia-induced apoptosis. To confirm the involvement of MAP kinase, JNK/SAPK, ERK, or p38 kinase assay was performed. Although p38 MAPK was activated in response to hypoxic treatment, the other MAPK -JNK/SAPK or ERK- was either only modestly activated or not at all. These results suggest that p38 MAPK is involved in hypoxia-induced apoptosis in MC3T3El osteoblasts.