• Applied Biological Chemistry
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• v.11
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• pp.1-27
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• 1969

More than thirty kinds of sea food pickles have been eaten in Korea. Out of these salted yellow tail pickle, salted clam pickle, salted oyster pickle, and salted cuttlefish pickle were employed for the analysis of their components, identification of main fermenting microbes, and determination of enzyme characteristics concerned. Also studied was the effect of enzymic action of microbes, which are concerned with the fermenting of pickles, on the production of flavorous 5'-mononucleotides and amino acids. The results are summarized as follows: 1. Microflora observed in the pickles are: (a) Total count of viable cells after 1-2 months of pickling was found to be $10^7$ and that after 6 months decreased to $10^4$. (b) Microbial occurence in the early stage of pickling was observed to be 10-20% Micrococcus spp., 10-20% Brevibacterium spp., 0-30% Sarcina spp., 20-30% Leuconostoc spp., ca 30% Bacillus spp., 0-10% Pseudomonas spp., 0-10% Flavobacterium spp., and 0-20% yeast. (c) Following the early stage of pickling, mainly halophilic bacteria such as Bacillus subtilis, Leuconostoc mesenteroides, Pediococcus halophilus and Sarcina litoralis, were found to exhibit an effect on the fermentation of pickle and their enzyme activities were in direct concern in fermentation of pickles. (d) Among the bacteria participating in the fermentation, Sarcina litoralis 8-14 and 8-16 strains were in need of high nutritional requirement and the former was grown only in the presence of purine, pyrimidine and cystine and the latter purine, pyrimidine and glutamic acid. 2. Enzyme characteristics studied in relation to the raw materials and the concerned microbes isolated are as follows: (a) A small amount of protease was found in the raw materials and 30-60% decrease in protease activity was demonstrated at 7% salt concentration. (b) Protease activity of halophilic bacteria, Bacillus subtilis 7-6, 11-1, 3-6 and 9-4 strains, in the complete media decreased by 10-30% at the 7% salt concentration and that of Sarcina litoralis 8-14 and 8-16 strains decreased by 10-20%. (c) Proteins in the raw materials were found to be hydrolyzed to yield free amino acids by protease in the fermenting microbes. (d) No accumulation of flavorous 5'-mononucleotides was demonstrated because RNA-depolymerase in the raw materials and the pickles tended to decompose RNA into nucleoside and phosphoric acid. (e) The enzyme produced in Bacillus subtilis 3-6 strain isolated from the salted clam pickles, was ascertained to be 5'-phosphodiesterase because of its ability to decompose RNA and thus accumulating 5'-mononucleotide. (f) It was demonstrated that the activity of phosphodiesterase in Bacillus subtilis 3-6 strain was enhanced by some components in the corn steep liquor and salted clam pickle. The enzyme activity was found to decrease by 10-30% and 40-60% at the salt concentration of 10% and 20%, respectively. 3. Quantitative data for free amino acids in the pickles are as follows: (a) Amounts of acidic amino acids such as glutamic and aspartic acids in salted clam pickle, were observed to be 2-10 times other pickles and it is considered that the abundance in these amino acids may contribute significantly to the specific flavor of this food. (b) Large amounts of basic amino acids such as arginine and histidine were found to occur in salted yellow tail pickle. (c) It is much interesting that in the salted cuttlefish pickle the contents of sulfur-containing amino acids were exceedingly high compared with those of others: cystine was found to be 17-130 times and methionine, 7-19 times. (d) In the salted oyster pickle a high content of some essential amino acids such as lysine, threonine, isoleucine and leucine, was demonstrated and a specific flavor of the pickle was ascribed to the sweet amino acids. Contents of alanine and glycine in the salted oyster pickle were 4 and 3-14 times as much as those of the others respectively. 4. Analytical data for 5'-mononucleotides in the pickles are as follows: (a) 5'-Adenylic acid and 3'-adenylic acid were found in large amounts in the salted yellow tail pickle and 5'-inosinic acid in lesser amount. (b) 5'-Adenylic acid, especially 3'-adenylic acid predominated in amount in the salted oyster pickle over that in the other pickles. (c) The salted cuttlefish pickle was found to contain only 5'-adenylic acid and 3'-adenylic acid. It has become evident from the above fact that clam and the invertebrate lack of adenylic deaminase and contain high content of adenylic acid. Thus, they were demonstrated to be the AMP-type. (d) 5'-Inosinic acid was contained in the salted yellow tail pickle in a significant concentration, and it might be considered to be IMP-type. 5. Comparative data for flavor with regard to the flavorous amino acids and the contents of 5'-mononucleotides are: (a) A specific flavor of salted yellow tail pickle was ascribed to the abundance in glutamic acid and aspartic acid, and to the existence of a small amount of flavorous 5'-inosinic acid. The combined effect of these components was belived to exhibit a synergistic action in producing a specific fiavor to the pickle. (b) A specific flavor of salted clam pickle has been demonstrated to be attributable to the richness in glutamic acid and aspartic acid rather than to that of 5'-mononucleotides.

• Journal of Sasang Constitutional Medicine
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• v.23 no.3
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• pp.391-401
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• 2011

1. Objectives: Though the eating habits have improved and the living method has become convenient according to the economic growth thanks to the modern industrialization, because of the lack of exercise, obesity, wrong eating habits and stress etc, various symptoms of disease of adults are on the rise. This is the phenomenon that happens as the eating life has become life in the West along with the inundation of the western culture in our society. In this perspective, there has been many various clinical research that's been proceeded so far about the physical constitution and obesity, but there has been little research on the objective analysis of the clinical research about the alimentotherapy using taeeumjoweetang. 2. Methods: In this research we have checked the weights, fat rates, fat weight, abdominal fat rate, blood pressure, and BMI over the objects of the women that were diagnosed as lunisolar system as their physical constitution, and assessed the paramecium lipid, in-blood antioxidation enzyme and the damage of oxidization in the urine by physical constitution-specific of the body shapes that were determined by BMI. The statistical analysis of the current research was processed by using of SPSS 17.0 program. We have figure out statistical amounts such as the arithmetic average, average deviation rate and percentage number. Fro the verification of he significancy of each elements, we have used the paired t-test, ANOVA, Chi-square test at the level of p<0.05. 3. Results and Conclusions: Their characteristics are age $21.20{\pm}1.35$, height $160.30{\pm}6.11cm$, weight $64.66{\pm}8.72kg$, fat rates are $35.97{\pm}4.87%$, fat amount $23.40{\pm}5.48$, abdominal fat rate $0.823{\pm}0.03$, BMI $25.12{\pm}2.79kg/m^2$, and systolic blood pressure $111.60{\pm}10.28mmHg$ and diastolic blood pressure $68.60{\pm}7.43mmHg$ and we have let them take the medicine twice a day for 8 weeks. The clinical result for the Cholesterol, Triglyceride, HDL-cholesterol, LDL-cholesterol, SGOT, SGPT, of the object people was that for the function of the liver, the result of the SGOT and SGPT test was $17.16{\pm}3.05$, $15.00{\pm}2.99IU/L$, which was a decrease, and had statistical meaning, but for the SGOT, though the figure reduced to $11.92{\pm}4.61$, $10.80{\pm}3.07$, it had no statistical meaning. For the whole cholesterol level, the figure reduced to $169.00{\pm}19.95$, $160.08{\pm}22.52$ mg/dL and had statistical significance(p<0.05). Neuter fat number, Triglyceride has slightly increased to $67.52{\pm}36.32$, $68.08{\pm}47.33$ mg/dL but did not have any statistical meaning. The antioxdant enzyme marker standard marker, SOD has increased to $2.52{\pm}0.73$, $2.86{\pm}0.60U/ml$, and had statistical significance(p<0.01). Catalase also increased by $0.63{\pm}0.18$, $1.07{\pm}0.25mmol/ml$ and had statistical meaning(p<0.01). GPx also increased to $204.76{\pm}32.64$ nmol/ml and had statistical meaning(p<0.01). But, for the Total antioxidant, though it has raised to $1.51{\pm}0.26$, $1.57{\pm}0.17nmol/{\mu}l$, it did not have any statistical meaning. MDA of oxidative stress marker has decreased to $1.70{\pm}0.68{\mu}g/ml$, $1.21{\pm}0.50{\mu}g/m$ and had statistical significance(p<0.01). 8-OHdG also decreased $3.35{\pm}0.95ng/ml$, $2.21{\pm}0.50ng/ml$ and had statistical meaning(p<0.01). In this research, we have analyzed the various markers relating to BFM and changes in oxidative enzyme in blood by takingtaeeumjoweetang. Taeeumjoweetang has the positive effect on inbody antioxidant system and reducing the content of cholesterol, which is proven to help losing weight and improving hyperlipidemia statistically. With this research, we hope to improve the lifestyle of those who are either obese or need to manage their dietary habits, and also to become the touchstone of integrating Oriental Medicine with the science of food & nutrition.

• Journal of Life Science
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• v.30 no.9
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• pp.826-833
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• 2020

Phospholipase C gamma (PLCγ) has critical roles in receptor tyrosine kinase- and non-receptor tyrosine kinase-mediated cellular signaling relating to the hydrolysis of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P₂] to produce inositol 1,4,5 trisphosphate (IP₃) and diacylglycerol (DAG), which promote protein kinase C (PKC) and Ca²⁺ signaling to their downstream cellular targets. PLCγ has two isozymes called PLCγ1 and PLCγ2, which control cell growth and differentiation. In addition to catalytically active X- and Y-domains, both isotypes contain two Src homology 2 (SH2) domains and an SH3 domain for protein-protein interaction when the cells are activated by ligand stimulation. PLCγ also contains two pleckstrin homology (PH) domains for membrane-associated phosphoinositide binding and protein-protein interactions. While PLCγ1 is widely expressed and appears to regulate intracellular signaling in many tissues, PLCγ2 expression is restricted to cells of hematopoietic systems and seems to play a role in the regulation of immune response. A distinct mechanism for PLCγ activation is linked to an increase in phosphorylation of specific tyrosine residue, Y783. Recent studies have demonstrated that PLCγ mutations are closely related to cancer, immune disease, and brain disorders. Our review focused on the physiological roles of PLCγ by means of its structure and enzyme activity and the pathological functions of PLCγ via mutational analysis obtained from various human diseases and PLCγ knockout mice.

• Journal of Life Science
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• v.24 no.2
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• pp.186-190
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• 2014

Carbonic anhydrase isozymes (CAs) are widespread zinc-containing metalloenzyme family. The enzyme catalyzes the reversible interconversion of $CO_2$ and $HCO_3$. This reaction is the main role of CA enzymes in physiological conditions. CA III, one of the CA isozymes, has been identified in many tissues. It is distinguished from the other isozymes by several characteristics, particularly by a lower specific activity and by its resistance to acetazolamide. However, the physiological function of CA III in fish is unknown. In this study, we examined the detection of CAs in the Shaggy sea raven Hemitripterus villosus, using SDS-PAGE, isoelectric focusing (IEF), and western blot analysis. We detected a significant protein band with molecular weight about 30 kDa from the tissues of H. villosus by SDS-PAGE and western blotting. A specific band of CA III with pI 7.0 was detected by IEF and western blotting in gill and muscle. The immunoreaction of anti-CA III expressed in the gill of H. villosus was much stronger than other tissues. One explanation for this result is that the fish gill is the only organ that is exposed to the external environment and that plays an important role in acid-base relevant ion transfer, the transfer of $H^+$ and/or $HCO{_3}^-$, for the maintenance of systemic pH. This is the first report on the identification of a carbonic anhydrase III-like protein from H. villosus.

• Biomedical Science Letters
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• v.4 no.1
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• pp.1-9
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• 1998

A Corynebacterium diphtheriae iron-repressible gene dirA, that was homologous to TSA of Saccharomyces cerevisiae and AhpC subunit of Salmonella typhimurium alkyl hydroperoxide reductase, was amplified with PCR and expressed in E. coli. The DirA purified from the transformed E. coli crude extracts prevented the inactivation of enzyme caused by metal-catalyzed oxidation (MCO) system containing thiols but not by ascorbate/Fe$^{3+}$/$O_2$ MCO system. The DirA concentration, which inhibited the inactivation of glutamine synthetase by 50% (IC$_{50}$) against MCO system, was 0.12 mg/ml. The multimeric forms of DirA were converted to the monomeric form in SDS-PAGE under the thioredoxin system comprised of NADPH, Saccharomyces cerevisiae thioredoxin reductase, and thioredoxin. Also, DirA showed thioredoxin dependent peroxidase activity. All of these results were consistent with the characteristics of a thiol specific antioxidant (TSA) protein having two conserved cysteine residues.

• Applied Biological Chemistry
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• v.42 no.4
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• pp.293-297
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• 1999

The pts operon, which encodes several factors in the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) of Escherichia coli, has multiple promoters which respond to different signals to facilitate quick adaptation to changes in growth conditions. The influence of an 1 kbp DNA region upstream of the pts P0 promoter on pts expression was studied in vitro by employing the DNA templates containing both P0 and P1 promoter with or without the 1 kbp upstream DNA region for in vitro transcription assay. The 1 kbp DNA region upstream of the pts P0 promoter, however, had no effect on pts transcription in vitro. The intracellular concentration of cAMP was measured when cells were grown in the presence of glucose, mannose, or mannitol. The transcription of P0 was increased maximally in the presence of glucose even though the concentration of cAMP in the condition was lowest while the transcription from the P1b was highest when cells were grown in the presence of mannose or mannitol even though the intracellular concentration of cAMP was lower than cells grown in the absence of the sugar. These results suggest the possibility of the existence of a glucose inducible repressor specific for the P0 promoter and a second repressor that is inducible by glucose, mannose and mannitol specific for the P1 promoter.

• Korean Journal of Food Science and Technology
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• v.16 no.2
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• pp.211-217
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• 1984

To study affinity of proteolytic enzymes to soy proteins, the physicochemical and functional properties of enzymatically modified protein products, kinetic parameters and degree of hydrolysis were measured using trypsin, alcalase (serine type protease) and pronase. Bacterial alcalase and pronase showed much greater affinity to soy protein than animal intestinal trypsin. This effect was very significant when unheated soy isolate was used as a substrate. Specific activities of these enzymes decreased with the increment of substrate concentration (over 2.0%, w/v) when heat denatured soy protein was used as a substrate. However, the decrease in specific activity was negligible at substrate concentrations lower than 2.0%. Polyacrylamide gel electrophoretic results showed that the pattern of 2S protein band changed distinctly in alcalase hydrolysis as compared with those of trypsin and pronase. Protein solubilities of alcalase and pronase hydrolyzates increased by 25-30%, at their pI (pH 5.0) over the control. Virtually no change was observed in solubility by trypsin hydrolysis. Heat coagulability and calcium-tolerance of the protein increased by enzymatic hydrolysis. No clear tendency, however, was observed for emulsion properties, foam expansion and the amount of free -SH groups. The enzyme treatment considerably decreased foam stability.

• Horticultural Science & Technology
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• v.28 no.4
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• pp.648-655
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• 2010

This study is the first comprehensive report on the molecular cloning, structural characterization, sequence comparison between wild and mutant types, copy number in the genome, expression features and activities of a gene encoding 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) in Korean lawn grass ($Zoysia$ $japonica$). The full length cDNA of the EPSPS from Korean lawn grass ($zj$EPSPS) obtained from a 3' and 5' RACE method was 1540 bp, containing a 1176 bp ORF, a 144 bp leader sequence (5' UTR) and a 220 bp 3' UTR, which was eventually decoded 391 amino acid residues with a molecular mass of 41.74 kDa. The Southern blot detection of the $zj$EPSPS showed that the gene exists as a single copy in the Korean lawn grass genome. Sequence comparison of the $zj$EPSPS gene demonstrated that the glyphosate-tolerant mutant (GT) having a Pro-53 to Ser substitution in the gene seems to have a preferred binding activity of the enzyme to phosphoenol pyruvate(PEP) over glyphosate, which allows the continuous synthesis of aromatic amino acids in the shikimate pathway. From the Northern blotting analysis, the $zj$EPSPS was found to be highly expressed, with continuous increase until 36 hours after 0.5% glyphosate treatment in both wild and mutant samples, but 1.5-fold higher EPSP synthase activity was observed in the tolerant mutant when exposed to the glyphosate treatment. The molecular information of the $zj$EPSPS gene obtained from this study needs to be further dissected to be more effectively applied to the development of gene-specific DNA markers and zoysiagrass cultivars; nevertheless, the glyphosate-tolerant mutant having the featured $zj$EPSPS gene can be provided to turfgrass managers for weed problems with timely adoptable management options.

• The korean journal of orthodontics
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• v.24 no.2
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• pp.303-317
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• 1994

This study was designed to measure the changes in the titer of tooth root antibodies accompanying root resorption associated with orthodontic tooth movement in dogs to explore a role of the specific immune response in root resorption during orthodontic tooth movement. Five adult mongrel dogs, 2 years of age, were used in the study. Six lower incisors were extracted as sources of homologous antigen in the dogs. Tooth root antigen preparations were made from a 6M Guanidine-HCl-10% EDTA(pH5.0) extract of these root dentins. Root resorption was elicited by intrusion of six maxillary incisors with 200-250gm intrusive force. In 9th week, resorbing six maxillary anterior teeth were extracted. Serum samples were taken from each dog prior to intrusion and weekly for 11 consecutive weeks. Serum autoantibody titers were determined with an enzyme-linked immunosorbent assay. As controls for antibody specificity, sera which were previously incubated with tooth root antigen as well as sera to an unrelated bacterial antigen (Porphyromonas gingivalis 33277) for 3 hours at 25 were measured in all runs. Root resorption was monitored monthly using occlusal radiographs. And then root resorption patterns were observed with a zoom stereo microscope (Model SZH-121, Olympus optical Co. Ltd.). Incisors did not show clear radiographic evidence of significant and progressive root resorption, but periodontal ligament space had widened. But root resorption was observed on the apical regions of the maxillary incisors with a zoom stereo microscope. Teeth showed the shallow depression generally accompanying deep resorption. These demonstrate a slight tendency for an immediate decrease followed by rebound to levels above the pre-treatment baseline. A peak titer of autoantibody to dentin antigen occurred on day 28, then steadily decreased during the 9th week period as the roots resorbed and then rapidly spiked in animals when the resorbing teeth were extracted. When sera is incubated with tooth root antigen, serum activity in the ELISA was almost absent. This is because serum activity in the ELISA could be removed by absorption of the serum with dog dentin antigen. Serum ELISA activity to the unrelated bacterial antigen remained essentially unchanged in all animals throughout the experimental period. When the time course of changes in autoantibody to homologous tooth root antigen prepatration and unrelated bacterial antigen was compared, no significant differences were found(${\alpha}=0.05$). In general, the overall pattern of changes in autoantibody was similar to the two antigens. These findings suggest the possibility that these immunologic changes precede a significant development of root resorption lesions rather than merely reflecting their presence. Therefore, this suggests that the changes of antibody levels may have some predictive value for root resorption.

• Applied Biological Chemistry
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• v.51 no.3
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• pp.159-163
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• 2008

Xylogone sphaerospora ${\beta}$-mannanase was purified by Sephadex G-100 column chromatography. The specific activity of the purified enzyme was 8.44 units/ml protein, representing an 56.27-folds purification of the original crude extract. The final preparation thus obtained showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 42kDa. Konjac glucomannan was hydrolyzed by the purified ${\beta}$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography. The main hydrolysates were composed of D.P. (Degree of Polymerization) 3 and 4 glucomannooligosaccharides. For elucidate the structure of D.P 3 and 4 glucomannooligosaccharides, sequential enzymatic action was performed. D.P 3 and 4 were identified as M-G-M and M-M-G-M (G- and M- represent glucosidic and mannosidic link-ages). To investigate the effects of konjac glucomannooligosaccharides on in vitro growth of Bifido-bacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum and B. breve. Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 3 and D.P. 4 glucomannooligosaccharides, respectively. B. longum and B. bifidum grew up 3.9-fold and 2.8-fold more effectively by the treatment of D.P. 4 glucomannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 4 was more effective than D.P. 3 glucomannooligosaccharide on the growth of Bifidobacterium spp.