• Journal of the East Asian Society of Dietary Life
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• v.7 no.3
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• pp.289-300
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• 1997

Newborn dog chymosin was extracted from the stomachs of dogs of 2 weeks of age, and was purified by ion exchange chromatography. Half of the sequence was determined by amino acid sequencing and the complete sequence was deduced from a cloned chymosin cDNA Results showed that the zymogen showed 79% sequence identity with calf prochymosin and 54% identity with porcine pepsinogen A The size of the propart and location of the residue which becomes the amino-terminus in the active enzyme was the same in the prochymosins. The maximum general proteolytic activity at pH 3.2 of newborn dog chymosin was 3-4% of that of porcine pepsin A at pH 2, whereas the milk clotting activity relative to the general proteolytic activity of newborn dog chymosin was much higher than that of calf chymosin. Agar gel electrophoresis at pH 5.2 of stomach extracts of individual dogs showed the existence of two predominant genetic variants of zymogen and enzyme. The two variants could not be distinguished by amino acid composition or amino-terminal sequencing, and no differences in the enzymatic properties of the genetic variants were observed. It was concluded that of the residues that participate in the substrate binding, calf and newborn dog chymosin differ in the following positions (porcine pepsin numbering, subsites in parentheses) : Ser 12 Thr(S$_4$), Leu 30 Val(S$_1$/S$_3$), His 74 Gln(S'$_2$), Val 111 Ile(S$_1$/S$_3$), Lys 220 Met(S$_4$). With regard to the low general proteolytic activity of newborn dog chymosin, the substitution Asp303 Val relative to calf chymosin may contribute to an explanation of this.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.3
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• pp.411-418
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• 2014

Three mushroom mycelia, Ganoderma lucidum, Hericium erinaceum, and Phellinus linteus, were separately diluted with the natural culture media Acanthopanax senticosus. Solid-state fermentation was used to produce three different A. senticosus-fermented mushroom mycelium groups: G. lucidum mycelia, H. erinaceum mycelia, and P. linteus mycelia. The resulting mycelia were analyzed to assess their efficacies as health functional foods. Optimized fermentation conditions were determined by considering the density and growth speed of mycelia in each A. senticosus-fermented mushroom mycelium group. The cultured mushroom mycelia under the optimized conditions were extracted using water and 70% ethanol. Extraction was followed by filtration, concentration and freeze-drying to produce extract powder of A. senticosus-fermented mushroom mycelia: Water extracts (FM-5111, FM-5121, and FM-5131) and 70% ethanol extracts (FM-5112, FM-5122, and FM-5132). Analysis of extract powder of A. senticosus-fermented mushroom mycelia was performed using the maker compounds eleutheroside B and eleutheroside E. Analysis of ${\beta}$-glucan contents was performed by enzymatic procedures.

• Journal of Life Science
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• v.25 no.6
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• pp.663-672
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• 2015

The study was performed to investigate the effects of enzyme treated garlic (EG) and its natural resources composites on lipid levels in serum and liver of rats fed a high fat diet. Four different types of EG-composite extracts prepared: EG and EG + grape peel (EGG), EG + Persimmon (EGP) and EG + Catechu (EGC) by mixed 9.5:0.5, 9:1 and 8:2 (w/w) ratios, respectively. DPPH and ABTS radical scavenging activity in vitro, show the highest in EG + Catechu (EGC) composite by mixed 8:2 (w/w). EG and EG-composites extracts (8:2, w/w) were administered orally to SD-male rats at a concentration of 2.5 g/kg/day for 5 weeks. Total lipid and cholesterol contents in serum were significantly lower in EGC group than control group, and triglyceride content was the lowest in EGP group by 54.29 mg/dL. HDL-cholesterol contents were significantly higher in EGP and EGC groups. LDL-cholesterol content was lower in EG group than EG-composite groups, and VLDL cholesterol content was the lowest in EGP group. GOT, GPT and ALP activity was significantly lower in EGP group. Total lipid, cholesterol and triglyceride contents in liver were significantly lower in EGP and EGC group than control group. Antioxidant activity in serum was the highest in EGC groups by 50.86%, in liver was the highest in EGP groups. TBARS content in serum and liver was the lowest in EGP group. In these results, we suggest that EGP composites could have hypolipidemic and anti-obesity effects in rats fed a high fat diet.

• Journal of Nutrition and Health
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• v.54 no.6
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• pp.618-630
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• 2021

Purpose: The ginger rhizome (Zingiber officinale) is widely cultivated as a spice for its aromatic and pungent components. One of its constituents, 6-hydroxydopamine (6-OHDA) is usually thought to cross the cell membrane through dopamine uptake transporters, and induce inhibition of mitochondrial respiration and the generation of intracellular reactive oxygen species (ROS). This study examines the neuroprotective effect and acetylcholinesterase (AChE) inhibitory activity of fermented ginger extracts (FGEs) on 6-OHDA induced toxicity in SH-SY5Y human neuroblastoma cells. Methods: Ginger was fermented using 2 species of Bacillus subtilis, with or without enzyme pretreatment. Each sample was extracted with 70% ethanol. Neurotoxicity was assessed by applying the EZ-Cytox cell viability assay and by measuring lactic dehydrogenase (LDH) release. Morphological changes of apoptotic cell nuclei were observed by Hoechst staining. Cell growth and apoptosis of SH-SY5Y cells were determined by Western blotting and enzyme activity analysis of caspase-3, and AChE enzymatic activity was determined by the colorimetric assay. Results: In terms of cell viability and LDH release, exposure to FGE showed neuroprotective activities against 6-OHDA stimulated stress in SH-SY5Y cells. Furthermore, FGE reduced the 6-OHDA-induced apoptosis, as determined by Hoechst staining. The occurrence of apoptosis in 6-OHDA treated cells was confirmed by determining the caspase-3 activity. Exposure to 6-OHDA resulted in increased caspase-3 activity of SH-SY5Y cells, as compared to the unexposed group. However, pre-treatment with FGE inhibited the activity of caspase-3. The neuroprotective effects of FGE were also found to be caspase-dependent, based on reduction of caspase-3 activity. Exposure to FGE also inhibited the activity of AChE induced by 6-OHDA, in a dose-dependent manner. Conclusion: Taken together, our results show that FGE exhibits a neuroprotective effect in 6-OHDA treated SH-SY5Y cells, thereby making it a potential novel agent for the prevention or treatment of neurodegenerative disease.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.6
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• pp.812-818
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• 2016

Protective effects of fermented Curcuma longa L. (CL) against alcoholic liver damage were investigated in HepG2/2E1 cells. Fermented CL was extracted by cold water (FCC), hot water, 80% ethanol, and methanol. Of the four extracts, the strongest hepatoprotective effect against ethanol-induced oxidative stress was observed in FCC. Pretreatment with FCC also reduced intracellular reactive oxygen species formation compared to ethanol-alone treated cells. FCC also enhanced catalase, glutathione-S-transferase, glutathione reductase, glutathione peroxidase, and superoxide dismutase, and non-enzymatic antioxidative activities such as glutathione compared to alcohol-treated HepG2/2E1 cells. Our findings suggest that FCC might be considered as a useful agent in the prevention of liver damage induced by oxidative stress by increasing the antioxidant defense mechanism.

• Tuberculosis and Respiratory Diseases
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• v.49 no.3
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• pp.310-322
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• 2000

Background : Phospholipase C(PLC) plays an important role in cellular signal transduction and is thought to be critical in cellular growth, differentiation and transformation of certain malignancies. Two second messengers produced from the enzymatic action of PLC are diacylglycerol (DAG) and inositol 1, 4, 5-trisphosphate (IP3). These two second messengers are important in down stream signal activation of protein kinase C and intracellular calcium elevation. In addition, functional domains of the PLC isozymes, such as Src homology 2 (SH2) domain, Src homology 3 (SH3) domain, and pleckstrin homology (PH) domain play crucial roles in protein translocation, lipid membrane modificailon and intracellular memrane trafficking which occur during various mitogenic processes. We have previously reported the presence of PLC-${\gamma}1$, ${\gamma}2$, ${\beta}1$, ${\beta}3$, and ${\delta}1$ isozymes in normal human lung tissue and tyrosine-kinase-independent activation of phospholipase C-${\gamma}$ isozymes by tau protein and AHNAK. We had also found that the expression of AHNAK protein was markedly increased in various mstologic types of lung can∞r tissues as compared to the normallungs. However, the report concerning expression of various PLC isozymes in lung canærs and other lung diseases is lacking. Therefore, in this study we examined the expression of PLC isozymes in the paired surgical specimens taken from lung cancer patients. Methods : Surgically resected lung cancer tissue samples taken from thirty seven patients and their paired normal control lungs from the same patients, The expression of various PLC isozymes were studied. Western blot analysis of the tissue extracts for the PLC isozymes and immunohistochemistry was performed on typical samples for localization of the isozyme. Results : In 16 of 18 squamous cell carcinomas, the expression of PLC-${\gamma}1$ was increased. PLC-${\gamma}1$ was also found to be increased in all of 15 adenocarcinoma patients. In most of the non-small cell lung cancer tissues we had examined, expression of PLC-${\delta}1$ was decreased. However, the expression of PLC-${\delta}1$ was markedly increased in 3 adenocarcinomas and 3 squamous carcinomas. Although the numbers were small, in all 4 cases of small cell lung cancer tissues, the expression of PLC-${\delta}1$ was nearly absent. Conclusion : We found increased expression of PLC-${\gamma}1$ isozyme in lung cancer tissues. Results of this study, taken together with our earlier findings of AHNAK protein-a putative PLD-${\gamma}$, activator-over-expression, and the changes observed in PLC-${\delta}1$ in primary human lung cancers may provide a possible insight into the derranged calcium-inositol signaling pathways leading to the lung malignancies.

• Applied Biological Chemistry
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• v.16 no.2
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• pp.60-77
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• 1973

The non-enzymatic browning phenomenons of red ginseng were studied to identify these compounds which function as the factors for browning. The samples were classified into five divisions; Fresh ginseng, blanched ginseng, sun dried red ginseng, dehydrated red ginseng, and browning accelerated red ginseng respectively, and the various compounds in each of them were analyzed quantitatively and investigated the compounds which were thought to function for browning during the drying and the dehydration processes; the results were as follows. 1. The chemical compositions among five divisions did not show any difference except a) total and reducing sugars, b) total acids, c) water soluble extracts; a) and b) were decreased during the drying process, c) was decreased about 6-7% in red ginseng divisions. 2. Sixteen free amino acids; asp., thr., ser., glu., gly., ala., val., cys., met., ileu., leu., tyr., phe., lys., his., and arg, were identified in each division. Among them the arg, was extremly high. All of the essential amino acids were contained, while generally these amino acids were decreased in drying period and their rates were smaller in dehydrated red ginseng than in sun dried red ginseng. 3. Three kinds of sugars; fructose, glucose and sucrose were identified and other four kinds of unidentified sugars were seperated. The content of sucrose was 80% and all kind of sugars were generally less in red ginseng divisions than in the other two divisions. The decreasing rate of sngars was higher in the sun dried red ginseng than in the dehydrated red ginseng. Especially the decreasing rate of the reducing sugars was high as compared with that of sucrose. 4. Almost all the ascorbic acid was decomposed during the blanching whereas there could'nt be shown any change of the ascorbic acid content during the period of drying. 5. Eleven kinds of volatile acids; acetic acid, propionic acid, acrylic acid, iso-butyric acid, n-butyric acid, isovaleric acid, n-valeric acid, isoheptylic acid, n-heptylic acid, and an unknown volatile acid were identified. They showed a little decrease during the period of blanching perhaps on account of their volatility whereas they were increased in drying period. 6. Six kinds of non-volatile acids; citric acid, malic acid, ${\alpha}-ketoglutaric$ acid, succinic acid, pyruvic acid and glutaric acid were identified. The content of them were decreased during the drying procedures in red ginseng but only that of succinic acid was increased. 7. Three kinds of polyphenols; 3-caffeyl quinic acid, 4-caffeyl quinic acid, 5-caffeyl quinic acid and an unknown polyphenol were identified. The content of them showed considerable decrease during the drying procedures, especially in sun drying. 8. The intensity of the browning in each divisior was as follows; browning accelerated red ginseng> sun dried red ginseng> dehydrated red ginseng. 9. In the process of red ginseng preparation, a. certain relationship could be found between the decreasing rates of amino acids, reducing sugars, polyphenols and the intensity of browning. Therefore the browning phenomenon may be concluded that nonenzymatic browning reactions of the amino-carbonyl reaction and autoxidation of polyphenols are the most important processes, furthermore, as their reactions could be controlled it is thought to be possible to accelerate effectively browning within a relatively short period.