• Journal of Plant Biotechnology
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• v.47 no.4
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• pp.283-288
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• 2020

Although the evolution of Arabidopsis thaliana and humans diverged approximately 1.6 billion years ago, recent studies have demonstrated that protein function and cellular processes involved in disease response remain remarkably conserved. Particularly, γ-secretase, a multisubunit protein complex that participates in intramembrane proteolysis (RIP) regulation, is also known to mediate the cleavage of more than 80 substrates including the amyloid precursor protein (APP) and the Notch receptor. Although the genes (PS1/2, APH-1, PEN-2, and NCT) coding for the γ-secretase complex components are present in plant genomes, their function remains largely uncharacterized. Given that the deposition of 42 amino acid long amyloid-β peptides (hAβ₄₂) is thought to be one of the main causes of Alzheimer's disease, we aimed to examine the physiological effects of hAβ₄₂ peptides on plants. Interestingly, we found that Arabidopsis protoplast death increased after 24 h of exposure to 3 or 5 µM hAβ₄₂ peptides. Furthermore, transgenic Arabidopsis plants overexpressing the hAβ₄₂ gene exhibited changes in primary root length and silique phyllotaxy. Taken together, our results demonstrate that hAβ₄₂ peptides, a metazoan protein, significantly affect Arabidopsis protoplast viability and plant morphology.

• Journal of Korean Medicine Rehabilitation
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• v.33 no.3
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• pp.17-32
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• 2023

Objectives The purpose of this study was to investigate the antioxidant, anti-inflammatory and cartilage regeneration effects of Euiiin-tang water extract (EIIT) in the treatment of monosodium iodoacetate (MIA)-induced osteoarthritis in rats. Methods Animal models were divided into five groups. The normal group didn't do any treatments causing osteoarthritis. The control group was orally administerd distilled water instead of the drug, the positive control group used indomethacin 5 mg/kg, the EIIT 100 group used EIIT 100 mg/kg and the EIIT 200 group used EIIT 200 mg/kg, and seven rats were placed per group. We administered drug to rats for 2 weeks and analyzed oxidative stress-related proteins in joint tissue. Inflammation mediators and inflammatory cytokines induced by the activity of inflammation-related proteins were analyzed. In addition, the expression of anti-inflammatory cytokines and collagen-related factors were analyzed, and H&E staining and Safranin-O staining were performed to see the effect on histopathological changes. Results 1) Oxidative stress-related proteins were significantly reduced. 2) Inflammationrelated proteins, inflammatory mediators and inflammatory cytokines were significantly reduced. 3) Anti-inflammatory cytokines were significantly increased. 4) Collagen proteolysis factors significantly decreased, and collagen degradation inhibitory factor was significantly increased. 5) EIIT administration significantly reduced cartilage degeneration and deformation in H&E staining, and reduced proteoglycan destruction in Safranin-O staining. Conclusions From the above experimental results, it judges that Euiiin-tang has antioxidant, anti-inflammatory, and cartilage regeneration effects on osteoarthritis in rats induced by MIA.

• Journal of Dairy Science and Biotechnology
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• v.41 no.3
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• pp.113-125
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• 2023

In this study, we explored the synergistic effects of whey protein concentrate (WPC) and soybean protein components after fermentation with lactic acid bacteria isolated from kimchi, and identified several peptides with desirable physiological functions, proteolysis, and immune effects. Antioxidant activity was determined using 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid, 1,1-diphenyl-2-picrylhydrazyl, ferric-reducing antioxidant power, and hydroxyl radical scavenging assays, followed by cross-validation of the four antioxidant activities. These assays revealed that samples with a 8:2 and 9:1 whey to soy ratio possessed higher antioxidant activity than the control samples. Antibacterial potency testing revealed high antibacterial activity in the 9:1 and 8:2 samples. Cytotoxicity testing of samples using 3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide revealed that only the 10:0, 1:9, and 0:10 samples had <80% viable cells, indicating no significant cytotoxicity. Nitric oxide (NO) assays revealed that NO expression was reduced in 8:2, 5:5, and 0:10 protein ratio fermentations, indicating low inflammatory reaction stimulatory potential. Cytokine expression was confirmed using an enzyme-linked immunosorbent assay kit. The 8:2 sample had the lowest inflammatory cytokine (interleukin [IL]-1α, IL-6, and tumor necrosis factor-α) levels compared with the lipopolysaccharide-treated group. Amino acid profiling of the 8:2 sample identified 17 amino acids. These results suggest that inoculating and fermenting Lactobacillus plantarum DK203 and Lactobacillus paracasei DK209 with an 8:2 mixture of WPC and soybean protein releases bioactive peptides with excellent anti-inflammatory and antioxidant properties, making them suitable for functional food development.

• Animal Bioscience
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• v.37 no.5
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• pp.908-917
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• 2024

Objective: Although pork loins is not a tough meat, they need to develop meat products with a soft texture for the elderly. This study focused on the physicochemical properties and tenderness characteristics of pork loin injected with green kiwifruit juice (GRJ) and gold kiwifruit juice (GOJ) during various incubation times. In addition, the antioxidant activities of hydrolysate derived from the hydrolysis of pork loin by kiwifruit juice protease were evaluated. Methods: The pork loin was injected with 10% and 20% GRJ and GOJ, under various incubation times (0, 4, 8, and 24 h). Then, the physicochemical properties and tenderness of pork loins were measured. 2,2- diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power were conducted to determine hydrolysate's antioxidant activities derived from pork loin's hydrolysis by kiwifruit juice protease. Results: GRJ had greater tenderizing ability than GOJ, even at the 10% addition. When kiwifruit juice was injected into pork loin, the tenderness increased with increasing incubation time. This was confirmed by the decrease in intensity of the myosin heavy chain (MHC) band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In particular, the MHC band decreased at 8 h for both 10% GRJ and 20% GOJ and at 4 h for 20% GRJ alone. The highest myofibril fragmentation index and peptide solubility were observed in pork loin treated with 20% GRJ compared to the other treatments during incubation. The 10% GRJ and 20% GOJ treatments showed similar levels of antioxidant activity of the protein hydrolysates in pork loin, and 20% GRJ showed the highest activity among the treatments. Conclusion: Kiwifruit juice had protease activity, and GRJ was more useful for tenderizing meat products than GOJ. Thus, GRJ at 10% could be a potential agent to tenderize and enrich the natural antioxidant activity through the proteolysis of pork loin.

• Journal of Life Science
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• v.19 no.9
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• pp.1226-1231
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• 2009

Enzyme substitution with recombinant phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is currently being explored for treatment of phenylketonuria (PKU), an autosomal recessive genetic disorder with mutations of the gene encoding phenylalanine-4-hydroxylase (EC 1.14.16.1). However, oral administration of PAL is limited because of proteolytic digestion in the gastrointestinal tract. The aim of this study was to determine the biochemical properties of PAL and delinate the susceptibility of wild-type PAL to pancreatic proteolysis by exploring several mutants, and to develop therapeutic drugs with PAL for PKU. The specific activity of PAL was assayed and its optimal pH, temperature stability, and intestinal protease susceptibility were investigated. Its $V_{max}$ values for phenylalanine and tyrosine were 1.77 and $0.47{\mu}mol$/ min/mg protein, respectively, and its $K_m$ values were $4.77{\times}10^{-4}$ and $4.37{\times}10^{-4}\;M$, respectively. PAL showed an optimal pH at 8.5, corresponding to the average pH range of the small intestine. It showed no loss of activity at $-80^{\circ}C$ for 5 months and possessed 93.4% of its activity under $4^{\circ}C$ for 4 wks. PAL was susceptible to chymotrypsin digestion and, to a lesser extent, to trypsin, elastase, carboxypeptidase A, and B. The trypsin and chymotrypsin cleaving sites were mutated to investigate protection from pancreatic digestion and the specific activities of these mutants were evaluated. The six mutants displayed low specific activities compared to the wild-type, suggesting that the primary trypsin and chymotrypsin cleaving sites may be essential for catalytic reaction. The PAL mutants could therefore be applied as a pretreatment modality without susceptibility to proteolytic attack, however, additional modification for enhancing enzymatic activity is needed to reduce the Phe levels effectively.

• Korean Journal of Food Science and Technology
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• v.15 no.2
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• pp.101-107
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• 1983

The four varieties of Korean soybeans were allowed to undergo natural fermentation for seven days at ambient temperature. The average pH of the product was 3.93 and titratable acidity was 1.94%. For all varieties of soybeans the content of riboflavin increased from 98 to $309.4{\mu}g/100g$ dry-matter, relative nutritive value from 78.66 to 94.59% and available lysine from 6.56 to 7.38 mg/gN, respectively. During fermentation, the activities of protease and lipase increased, while lipoxygenase and trypsin inhibitor activity decreased markedly. The capacity of water sorption of fermented soybean flour was increased with progress of proteolysis during fermentation. The cookie and noodle prepared with 20:80 mixture of fermented soybean flour and wheat flour were in the 'like' category, but it was desirable to neutralize the sour taste produced by fermentation. Among five kinds of products prepared from the fermented soybean flour pan cake was liked most by rural consumers.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.1
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• pp.172-180
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• 2002

Polychlorinated biphenyls(PCBs) are large scale industrial chemicals which are using in diverse applications. The goal of this study was to determine if exposure to 2,2',5,5'-tetrachlorobiphenyl (PCB 52) leads to an increase in the production of active oxidants, and subsequently promotes apoptosis of neuronal SK-N-MC cells. Reactive oxygen species (ROS) formation was examined in SK-N-MC cells after treatment of PCB 52 by concentrations and incubation times, respectively. It showed that the rate of ROS production in the cells was increased in a does-dependent manner to 45 min, followed by a return towards control levels after 120 min treatment. We also examined the association of PCB-induced apoptosis with the modulation of biomakers of oxidative damage to lipids (malondialdehyde [MDA]) in SK-N-MC cells. Increased MDA was observed in a dose-dependent manner in groups treated with 10, 15, and 20 figJ me of PCB 52 for 24 h. After treatment of PCB 52, the cells did not show any significant change in the rate of Cu/Zn-superoxide dismutase (Cu/Zn-SOD) activity. Whereas, the cells had a two-fold greater rate of change in catalase activity at 20 ㎍/㎖ of PCB 52 for 24 h when compared to control group. Korean Ginseng is one of the most important crude drugs which has been used as a traditional Oriental medicine. We next investigated protective effect of extracts of ginseng on cytotoxicity induced by PCB 52 in SK-N-MC cells. Pretreatment of SK-N-MC cells with 25-200 μg/ml of ginseng were reduced cell death in a dose-dependent manner in PCB 52-treated cells. To examine the sensitivity of beta-catenin to ginseng, the protective effect of a range of ginseng concentrations was examined in SK-N-MC cells treated with PCB 52. The result demonstrated that ginseng efficiently blocked PCB 52 inducible beta-catenin proteolysis in a concentration dependent manner. The ROS formation was also measured in the presences of extract of ginseng and superoxide dismutase (inhibitor of oxygen free radical production). The both SOD (400 U/ml) and ginseng (200 μg/ml) significantly inhibited RDS generation in PCB 52-treated group.

• Journal of Oriental Neuropsychiatry
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• v.26 no.1
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• pp.63-78
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• 2015

Objectives: Pinelliae Rhizoma has traditionally been used as an anti-depressant in oriental medicine. This study is to investigate the effect of Pinelliae Rhizoma extract (PRe) on psychological stress in genome wild expression of mice. Methods: After giving physical stress to mice, PRe was orally administered with 100 mg/kg/day for five days. After extracting whole brain tissue from the mice, their genome changes were observed by micorarray analysis method. The genome changes were analyzed by IMAGENE 4.0, TREEVIEW, FatiGo algorithems, BOND database, cytoscape program, etc. Results: 1. PRe administered group were remained at normal level; 60% of increase was shown in expressed genes by physical stress, and 65% of decrease was shown in expressed genes by psychological stress. 2. Genes with increased expression in control group that remained at a normal state in PRe administered group were involved with the gene of a cellular metabolic process on biological process, protein binding on molecular function, and cell part on cell composition. The pathway was found to be cytokin-cytokin receptor interaction. 3. Genes with decreased expression in control group that remained at a normal state in PRe administered group were involved with the gene of a cellular metabolic process on biologiacl detail and coupled ATPaes activity on molecular function. This gene related path was Ubiquintin mediated proteolysis etc. 4. Core node genes analyzed by protein interaction network were Vinculin, Cell sdivision cycle 42 homolog (S. cerevisiae) etc. They played an important role in maintaining cytoskeleton and controlling cell cycle. Conclusions: Several genes were up-regulated and down-regulated in response to psychological stress. The expression of most of the genes that were altered in response to psychological stress was restored to normal levels in PRe treated mice. When the interaction network information was analyzed, the recovery of the core node genes in PRe treated mice indicates that this final set of genes may be the effective target of PRe.

• Analytical Science and Technology
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• v.24 no.6
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• pp.510-518
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• 2011

Recent developments and improvements of multiple technological elements including mass spectrometry (MS) instrument, multi-dimensional chromatographic separation, and software tools processing MS data resulted in benefits of large scale proteomics analysis. However, its throughput is limited by the speed and reproducibility of the protein digestion process. In this study, we demonstrated a new method for rapid proteolytic digestion of proteins using acoustic technology. Tryptic digests of BSA prepared at various conditions by super acoustic for optimization time and intensity were analyzed by LC-MS/MS showed higher sequence coverage in compared with traditional 16 hrs digestion method. The method was applied successfully for complex proteins of a breast cancer cells at 30 min of digestion at intensity 2. This new application reduces time-consuming of sample preparation with better efficiency, even with large amount of proteins, and increases high-throughput process in sample preparation state.

• Journal of Life Science
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• v.17 no.9 s.89
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• pp.1224-1231
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• 2007

Insulin-like growth factor-I (IGF-I) and IGF-II have structure like insulin. In contrast to insulin, however, the bioavaility of IGFs is modulated by the IGF-binding protein (IGFBPs). Each of IGFBPs was different with molecular masses, biological characteristics, and immunological properties.. Human fibroblasts secrete IGFBPs that can modify IGF-I action. In diabetes mellitus, the most study of IGF systems have been investigated in insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus, and streptozotocin-in-duced animals in vivo. Recently, a little research regarding the IGFs system has been proposed in por-tion of cell in vitro. In this study, effects of low or high glucose condition on IGFBP-5 in GM10 was investigated. By western blotting analysis, IGFBP-5 level decreased in cells cultured at high glucose, but IGFBP-5 level of mRNA didn't change. IGFBP-5 protease that cleaves IGFBP-5 in conditioned me-dium had was inhibited by EDTA and heparin, like serine protease and metalloprotease. Furthermore, the protease activity was increased in high glucose cultivated condition. In results of gelatin zymog-raphy, molecular weight of proteolytic metalloenzymes was indentified 69-kDa and protease activity was increased in time-dependent manner. Although the mechanism has yet to be determined, IGFBP-5 proteolysis in GM10 cells cultured with high glucose may increase effects of IGFs to decrease the glu-cose level through dissociation of IGFs from IGFBPs. Therefore, we suggest that IGF- I and IGFBPs could be potential models in study of pathophysiology such as diabetes mellitus.