• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.5
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• pp.509-514
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• 1994

A processing method for fermented paste of favorable flavor and texture using pen shell by-product was investigated. The pen shell by-product was homogenized with the addition of water and hydrolyzed with $5\%$ of Protin P(105 PU/g) at $55^{\circ}C$ for 1 hour. Flavor of the hydrolysate could be improved by thermal treatment at $100^{\circ}C$ for 40 minutes with $10\%$ of invert sugar. $2\%$ of agar-agar and $6\%$ of starch added to hydrolysate emulsified by $8\%$ of polyglycerol condensed ricinoleate(PGDR) were significantly effective for the improvement of rheological properties such as hardness, springiness and chewiness of the fermented paste. $15\%$ of table salt was finally added to the product of fermented pen shell paste. The contents of moisture, protein, lipid, carbohydrate and salinity of the product were $62.7\%,\;3.2\%,\;4.4\%,\;10.6\%\;and\;15.6\%$, respectively. The major free amino acids were glutamic acid, arginine, aspartic acid, leucine, lysine, glycine and alanine. The product was stable for the storage of 60 days at $23{\pm}3^{\circ}C$ on bacterial growth.

• Applied Biological Chemistry
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• v.15 no.2
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• pp.93-109
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• 1972

Three lots of Chung-Kook-Jang were prepared by the use of 2 strains of Bacillus subtilis and Bacillus natto. For four samples taken from each lot in 12 hrs interval changes of nitrogenous compounds, insoluble protein, water soluble protein, peptides, free amino acids, amino and ammonia nitrogens during Chung-Kook-Jang fermentation, were studied together with the changes of moisture, pH, proteolytic enzyme activity. In addition the average peptide length of the peptides of a Bacillus subtilis lot was determined by the method of molecular sieving using ion exchange resin. The results were as follows: 1. The contents of moisture and total-nitrogen changed little in all samples throughout the fermentation as it would be expected. 2. In all three experimental lots the pH became higher gradually from the initial value of 6.65 to the final $7.5{\sim}7.85$ during the fermentation. Proteolytic enzyme activities, in accordance with this pH change, steadily increased up to $48{\sim}60$ hrs. of fermentation and then slightly decreased, probably affected by the high pH. The most strong proteolytic activity was observed in the experimental Chung-Kook-Jang fermentation lot using the Bacillus subtilis K-27 isolated by the author. 3. The contents of insoluble protein nitrogen in soybeans increased markedly (5%) by the cooking, after steeping 12 hrs in water. During the Chung-Kook-Jang fermentation, however, it decreased from 1/2 to 1/10 of that of the cooked soybeans. 4. The contents of water soluble protein nitrogen (5%) whereas, greatly decreased to the value of 1.0% by the cooking; but little changed further during the fermentation, 5. The total contents (0.25%) of peptides, amino, and ammonia-nitrogens, PAA-N., increased almost double by the cooking and steadily became higher as the fermentation proceeded, reaching finally up to$4{\sim}7%$ in 72 hrs fermentation. 6. The amounts of free amino acids of soybean generally decreased during the processing of cooking, even some of them like glutamic acid were destroyed completely, However in the subsequent Chung-Kook-Jang fermentation for 72 hrs., they showed from several to a few hundreds folds increases depending upon the kinds of amino acids. Valine which was contained in HCl-hydrolyzed steeped or cooked soybeans in amounts $220{\sim}267mg%$ was not detected at all as the free amino acid in all fermented samples. 7. Average peptide length (APL) of all fractions, eluted and fractionated by using the Dowex-50 ion exchange resin column, and fraction collector showed the highest value for the cooked soybean and then decreased as the fermentation proceeded. The APL value of effluent showed the highest in 12 hrs fermented sample, The value decreased thereafter by fermentation.

• Journal of Life Science
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• v.22 no.4
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• pp.552-558
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• 2012

Severe acute respiratory syndrome (SARS) is a severe respiratory infectious disease caused by a novel human coronavirus, SARS-CoV. The 3CL protease is a key enzyme in the proteolytic processing of replicase polyprotein precursors, pp1a and pp1ab, which mediate all the functions required for viral genomic replication and transcription. Therefore, this enzyme is a target for the development of chemotherapeutic agents against SARS. A large quantity of active SARS-3CL protease is required for development of anti-SARS agents. Here we have constructed overexpression vector for the production of the SARS-3CL protease. The gene encoding SARS-3CL protease was amplified using polymerase chain reaction and cloned into the pET29a expression vector, resulting in pET29a/SARS-3CLP. Recombinant SARS-3CL protease was successfully synthesized by the dialysis mode of the cell-free protein expression system, and purified by three-step fast protein liquid chromatography using HighQ and MonoP column chromatographies and Sephacryl S-300 gel filtration. In addition, the produced SARS-3CL protease was found to be an active mature form. This study provides efficient methods not only for the development of anti-SARS materials from natural sources, but also for the study of basic properties of the SARS-3CL protease.

• 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.

• The Journal of Korean Medicine
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• v.29 no.2
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• pp.151-164
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• 2008

Objective: To investigate the effects of Yeongdamsagantang (YDGT) on apoptosis of neuronal cells that can result in dementia. Method: The water extract of the YDGT was tested in vitro for its beneficial effects on neuronal survival and neuroprotective functions, particularly in connection with $A{\beta}$ oligomer-related dementias. $A{\beta}$ oligomers derived from proteolytic processing of the ${\beta}-amyloid$ precursor protein (APP), including the $amyloid-{\beta}$ peptide $(A{\beta})$, play a critical role in the pathogenesis of Alzheimer's disease. A neuroblastoma cell line stably expressing an $A{\beta}$ oligomerassociated neuronal degeneration was used to investigate if YDGT inhibits formation of $A{\beta}$ oligomer. To measure the ATP generating level in mitochondrial membrane, luciferin/luciferase luminescence kit (Promega) and luminator was used, and to survey the protein's apparition, confocal microscopy was used. Result: $A{\beta}oligomer$ had a profound attenuation in the increase in CT105 expressing neuro2A cells from YDGT. Experimental evidence indicates that YDGT protected against neuronal damage from cells, but its cellular and molecular mechanisms remain unknown. We demonstrated that YDGT inhibited formation of $amyloid-{\beta}$ $(A{\beta})$ oligomers, which were the behavior, and possibly causative, features of AD. The decreased $A{\beta}$ oligomer in the presence of YDGT was observed in the conditioned medium of this $A{\beta}oligomer-secreting$ cell line under in vitro. In the cells, YDGT significantly attenuated mitochondrion-initiated apoptosis. Conclusion: (i) a direct $A{\beta}$ oligomer toxicity and the apoptosis initiated by the mitochondria; and (ii) multiple cellular and molecular neuroprotective mechanisms, including attenuation of apoptosis and direct inhibition of $A{\beta}$ oligomer aggregation, underlie the neuroprotective effects of YDGT.

• Food Science and Preservation
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• v.23 no.3
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• pp.291-300
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• 2016

Kiwifruit (Actinidia chinensis, Hayward) was stored at $25^{\circ}C$ for 0~30 days and investigated to find out the optimum storage time to obtain the best physical and functional properties for consumers' preference. Kiwifruits was stored at different time period (0, 5, 10, 15, 20, and 30 days) for investigating their physiochemical quality, nutritional components, and functional characteristics. Kiwifruits stored for 20~30 days showed the best physiochemical quality such as higher total acidity and proper firmness. They were also more enriched with dietary fibers, free sugar, and organic acid, although no significant changes were observed in crude protein, crude fat, and moisture content. For functional properties, kiwifruits stored for 20 days showed significantly higher contents of total phenolics, flavonoids, and actinidin. In addition, it showed stronger antioxidant activity, whitening effect, and proteolytic activity when compared with other samples. SDS-PAGE analysis showed the presence of actinidin enzyme in kiwifruits. These results indicated that the kiwifruits stored for 15~20 days possessed excellent quality and high concentrations of nutritional and functional compounds, which could be best for both fresh consumption and product processing.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.2
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• pp.397-404
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• 2009

For standardization of LMK02 quality, Ginsenoside Rg3 of Red Ginseng and Decursin of Angelica gigas Nakai in the constituents of LMK02 were estimated as indicative components. From LMK02 water extract, has been used in vitro test for its beneficial effects on neuronal survival and neuroprotective functions, particularly in connection with APP-related dementias and Alzheimer's disease (AD). $A{\beta}$ oligomer derived from proteolytic processing of the ${\beta}$-amyloid precursor protein (APP), including the amyloid-${\beta}$ peptide ($A{\beta}$), play a critical role in the pathogenesis of Alzheimer's dementia. We determined that oligomer amyloid-${\beta}$ ($A{\beta}$) have a profound attenuation in the increase in rat hippocampus H19-7 cells from. Experimental evidence indicates that LMK02 protects against neuronal damage from cells, but its cellular and molecular mechanisms remain unknown. Using a hippocampus cell line on $A{\beta}$ oligomer-induced neuronal cytotoxicity, we demonstrated that LMK02 inhibits formation of $A{\beta}$ oligomer, which are the behavior, and possibly causative, feature of AD. In the Red Ginseng, the average amounts of Ginsenoside Rg3 were $47.04{\mu}g/g$ and $42.3{\mu}g/g$, 90 % of its weight were set as a standard value. And, in the Angelica gigas Nakai, the average amounts of Decursin were 2.71 mg/g and 2.44mg/g, 90 % of its weight were also set as a standard value. The attenuated $A{\beta}$ oligomer in the presence of LMK02 was observed in the conditioned medium of this $A{\beta}$ oligomer-induced cells under in vitro. In the cells, LMK02 significantly activated antiapoptosis and decreased the production of ROS. These results suggest that neuronal damage in AD might be due to two factors: a direct $A{\beta}$ oligomer toxicity and multiple cellular and molecular neuroprotective mechanisms, including attenuation of apoptosis and direct inhibition of $A{\beta}$ oligomer, underlie the neuroprotective effects of LMK02 treatment.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1653-1658
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• 2002

The objectives of this study were to examine calpain activity and meat tenderness by three different feeding patterns in Korean native cattle (KNC). Total forty-five animals were assigned each fifteen in long term restriction feeding (LTFR), long-term restriction feeding and hormone treatment (LTFR-tH), and short term non-restriction feeding (STFNR), respectively. Concentrate was restricted based on body weight in exp 1 and 2. However, it was fed ad libitum in exp. 3. Hormonal implantation was made with $M-PO^{TM}$ for bulls and with $F-TO^{TM}$ for heifers at 18, 20, 22 months of age in exp. 2. Animals were purchased (3-5 month old) from local cattle market and managed in two local farms and university research unit at three different years. Animals were slaughtered at 24 months for long-term trial and at 18 month for short term-trial. Loin and tender loin muscle was used for calpain activity and meat quality. Calpain proteolytic system was not changed by treatment. However, calpastatin activity was low in short-term trial. The calpain and calpastatin activity is reciprocal relationship, therefore, the high calpain activity may effect on quality grade. The shear force value was decreased as the processing of aging after postmortem. On the other hand, the cooking loss was significantly higher in short-term than in long-term trial, and then gradually decreased by the aging. Hormone implants to increase meat yield influenced to calpastatin activity more powerfully than calpain activity to meat tenderness. In meat color-a*, there was not significant difference in loin. Meat color-b* was decreased as postmortem aging time increased in tenderloin. Western blots were done to learn whether these proteins are degraded during postmortem storage and whether this degradation temporally parallels the decrease of shear force value. Vinculin was detected at 0 day and 1 day and degraded after 3 day. In conclusion, Calpain activity was affected slightly on meat tenderness. But meat tenderness was influenced by calpastatin, more effectively.

• The Korean Journal of Nuclear Medicine Technology
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• v.13 no.1
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• pp.116-119
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• 2009

Background: Preanalytical factors can affect reliability of hormone assay results. Adrenocorticotropic hormone (ACTH) in blood is considered highly unstable because of proteolytic degradation, so storage of blood samples on ice until analysis is recommended. In clinical practice, however, this procedure may present logistical problems because most samples for ACTH measurement must be shipped from the place of sample collection to the laboratory. Therefore, we studied the impact of time and temperature before plasma separation and analysis on the results of ACTH assays. Methods: A total number of 22 patients were enrolled in this study. We obtained 2 blood samples. ACTH concentrations were 35~126 pg/mL. ACTH concentrations were measured by immunoradiometric assay (IRMA) using commercial kits (CIS Biointernational, Gif-sur-Yvette, France). Results: ACTH levels showed a significant difference between the samples of $22^{\circ}C$ EDTA and $4^{\circ}C$ EDTA. Measured ACTH concentrations significantly decreased with time before freezing at $-20^{\circ}C$. ACTH levels showed no significant difference between the groups of after storage for 24 hr without centrifugation at $22^{\circ}C$ and $4^{\circ}C$. Conclusion: We recommend that blood samples be obtained on pre-chilled EDTA collection tubes. The shortest possible time between sample collection and processing is always the best laboratory practice.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.1
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• pp.8-17
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• 2011

To identify and examine the distribution of proteolytic inhibitory activity in crude extracts from fish eggs, and to determine the applicability of these protease inhibitors as anti-degradation agents in surimi-based products and fish meat, we compared the inhibitory activities of various extracts from fish eggs to those of commercial proteases, such as trypsin and papain. We used the optimal conditions for the screening of trypsin activity: 30 ug/uL of 0.1% trypsin and 0.6 mM Na-benzoyl-L-arginine-p-nitroanilide (BAPNA) with a pH of 8.0 at $40^{\circ}C$ for 60 min. The activities of papain and four commercial proteases were investigated after mixing with 100 ug/uL enzymes and 0.3% casein with a pH of 8.0 at $40^{\circ}C$ for 60 min. We performed a screening assay to detect the inhibitory activity (%) of crude extracts from eight species of fish eggs against the target proteases trypsin and papain. The assay revealed a wide distribution of trypsin and papain inhibitors in fish eggs. The specific inhibitory activities (11.6.28.6 U/mg) of crude extracts from fish eggs against trypsin and BAPNA substrate were higher than that (0.64 U/mg) of egg whites, used as a commercial inhibitor. The inhibitory activities of crude extracts from fish eggs against trypsin, and of egg whites against casein substrate (1.94.4.51 U/mg), were higher than those of papain (0.24.1.57 U/mg) and commercial protease (0.04.0.32 U/mg). The extracts from fish eggs were rich in protease inhibitors that exhibited strong inhibitory activity against trypsin, a serine protease, and papain, a cysteine protease.