• Archives of Pharmacal Research
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• v.28 no.7
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• pp.816-822
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• 2005

Catharsius protease-2 (CPM-2) was isolated from the body of dung beetles, Catharsius molossus, using a three step purification process (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and affinity chromatography on DEAE Affi-Gel blue). The purified CPM-2, having a molecular weight of 24 kDa, was assessed homogeneously by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of CPM-2 was composed of X Val Gin Asp Phe Val Glu Glu lie Leu. CPM-2 was inactivated by $Cu^{2+}\;and\;Zn^{2+}$ and strongly inhibited by typical serine proteinase inhibitors such as TLCK, soybean trypsin inhibitor, aprotinin, benzamidine, and ${\alpha}_1$-antitrypsin. However, EDTA, EGTA, cysteine, $\beta$-mercaptoethanol, E64, and elastatinal had little effect on enzyme activity. In addition, antiplasmin and antithrombin III were not sensitive to CPM-2. Based on the results of a fibrinolytic activity test, CPM-2 readily cleaved $A{\alpha}-$ and $B{\beta}$-chains of fibrinogen and fibrin, and y-chain of fibrinogen more slowly. The nonspecific action of the enzyme resulted in extensive hydrolysis, releasing a variety of fibrinopeptides of fibrinogen and fibrin. Polyclonal antibodies of CPM-2 were reactive to the native form of antigen. The ELISA was applied to detect quantities, in nanograms, of the antigen in CPM-2 protein.

• Korean Journal of Microbiology
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• v.16 no.3
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• pp.122-130
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• 1978

During the sporulation stage in Saccharomyces cerevisiae J170, the incorporation of $D^{14}$ C-glucose into starved cells of sporulation stage as well as the vegetative one is appeared higher at pH 6.0. Glucose transport system, in both the vegetative and sporulation stage, is associated with "energy dependent" as the result of repression by such a respiratory inhibitor as 2, 4-dinitrophenol. The Km value of glucose uptake in vegetative stage and sporulation stage was 2.1 mM and 2.5 mM respectively, indicating that the glucose is considerably reuqired for vegetative growth. Competition and countertranspoer of glucose by frutose and galactose are more distinct in vegetative stage, comparing with sporulation stage. The main sugar components of yeast cells consists of ribose, mannose, and ${\alpha}, \;{\beta}-glucose$. Amounts of mannose is lower in the aporulation stage than that in the vegetative stage.

• Journal of the Korean Society of Food Science and Nutrition
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• v.12 no.4
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• pp.387-394
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• 1983

The factor, which was capable of accelerating the yeast cell wall lysis of the zymolyase(${\beta}-1$, 3-glucanase), was purified to a homogeneous state from the protease fraction of the crude zymolyase by Sephadex G-75 gel filtration and preparative polyacrylamide gel disc electrophoresis. The molecular weight of the purified factor was estimated to be 40,500 by SDS-polylacrylamide gel disc electrophoresis and it's iso-electric point was pH 9.6. The factor was found to be a basic protease consisted of single polypeptide chain with 395 amino acid residues and it showed the $E_{280,cm}^{1%}$ of 11.9 and the molecular extinction coefficient of $4.83{\times}10^4$, respectively.

• Journal of Microbiology and Biotechnology
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• v.29 no.8
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• pp.1212-1220
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• 2019

The study of metabolomics in natural products using the diverse analytical instruments including GC-MS, LC-MS, and NMR is useful for the exploration of physiological and biological effects and the investigation of drug discovery and health functional foods. Cordyceps militaris has been very attractive to natural medicine as a traditional Chinese medicine, due to its various bioactive properties including anti-cancer and anti-oxidant effects. In this study, we analyzed the metabolite profile in 50% ethanol extracts of C. militaris fruit bodies from three development periods (growth period, matured period, and aging period) using $^1H-NMR$, and identified 44 metabolites, which are classified as 16 amino acids, 10 organic acids, 5 carbohydrates, 3 nucleotide derivatives, and 10 other compounds. Among the three development periods of the C. militaris fruit body, the aging period showed significantly higher levels of metabolites including cordycepin, mannitol (cordycepic acid), and ${\beta}-glucan$. Interestingly, these bioactive metabolites are positively correlated with antitumor growth effect; the extract of the aging period showed significant inhibition of HepG2 hepatic cancer cell proliferation. These results showed that the aging period during the development of C. militaris fruit bodies was more highly enriched with bioactive metabolites that are associated with cancer cell growth inhibition.

• Food Science and Preservation
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• v.23 no.5
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• pp.680-688
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• 2016

This study analyzed the changes in physicochemical components of Astragalus membranaceus (AM) fermented with Phellinus linteus. Moisture content, pH, total acidity, total reducing sugar content, extraction yield, free sugar content, free amino acid and isoflavonoid (calycosin, formononetin) were investigated. The moisture content was increased during fermentation with Phellinus linteus. The pH level increased while the total acidity significantly decreased during fermentation. The reducing sugar content were in the range of 0.32~0.61%. The extraction yield using water was higher than that using 80% ethanol. The major free sugars were identified as glucose, fructose, sucrose and the content of free sugars decreased through fermentation. However, the glucose and sucrose contents of the water extracts were increased. In addition, the free amino acid increased significantly during fermentation. Finally, calycosin and formononetins contents in water extracts of after 30 days of AM fermentaion with Phellinus linteus were (3.91 mg/100 g) and (1.38 mg/100 g), respectively. These results suggest that fermentation with Phellinus linteus could be used to increase the bioactivity of AM. The mycelium-fermented AM could be a valuable source of functional material and edible resource for industry.

• Korean Journal of Food Science and Technology
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• v.48 no.4
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• pp.306-316
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• 2016

This study was conducted to evaluate the functional characteristics and safety properties of lactic acid bacteria isolated from salt-fermented anchovy, a putative probiotic candidate. The following isolates were identified by biochemical profiles, carbohydrate fermentation patterns, and 16S rRNA sequencing: Enterococcus faecium AJ06, Leuconostoc mesenteroides AJ13, Pediococcus halophilus AJ22, Lactobacillus sakei AJ29, and Pediococcus pentosaceus AJ35. The strains AJ06, AJ22, AJ29 exhibited high tolerance to simulated gastric and intestinal juices and were able to produce bile salt hydrolase on MRS agar plates supplemented with taurocholic acid and/or taurodeoxycholic acid. The strains AJ22 and AJ29, which demonstrated high adherence to Caco-2 cells and resistance to various antibiotics, effectively inhibited the growth of food-borne pathogens by the production of antimicrobial substances. These strains did not show ${\alpha}-$ or ${\beta}$-haemolysis on blood agar. Furthermore, biogenic amines in MRS broth containing the precursor amino acids were not mutagenic in Salmonella Typhimurium TA98 and TA100.

• BMB Reports
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• v.39 no.4
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• pp.346-354
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• 2006

The full-length cDNA of grass carp (Ctenopharyngodon idellus) and silver carp (Hypophthalmichthys molitrix) uncoupling protein 2 (UCP2) was obtained from liver. The grass carp UCP2 cDNA was determined to be 1152 bp in length with an open reading frame that encodes 310 amino acids. Five introns (Intron 3, 4, 5, 6 and 7) in the translated region, and partial sequence of Intron 2 in the untranslated region of grass carp UCP2 gene were also obtained. Gene structure comparison between grass carp and mammalian (human and mouse) UCP2 gene shows that, the UCP2 gene structure of grass carp is much similar to that of human and mouse. Partial UCP2 cDNA sequences of bighead carp (Aristichthys nobilis) and mud carp (Cirrhinus molitorella), were further determined. Together with the common carp (Cyprinus carpio) UCP2 sequence from GenBank (AJ243486), multiple alignment result shows that the nucleotide and amino acid sequences of the UCP2 gene, were highly conserved among the five major Chinese carps that belong to four subfamilies. Using beta-actin as control, the ratio UCP2/beta-actin mRNA (%) was determined to be $149.4{\pm}15.6$ (common carp), $127.4{\pm}22.1$ (mud carp), $96.7{\pm}12.7$ (silver carp), $94.1{\pm}26.8$ (bighead carp) and $63.7{\pm}16.2$ (grass carp). The relative liver UCP2 expression of the five major Chinese carps, shows a close relationship with their food habit: benthos and detrituseating fish (common carp and mud carp) > planktivorious fish (silver carp and bighead carp) > herbivorious fish (grass carp). We suggest that liver UCP2 might be important for Chinese carps to detoxify cyanotoxins and bacteria in debris and plankton food.

• Microbiology and Biotechnology Letters
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• v.41 no.4
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• pp.391-397
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• 2013

A putative cyclomaltodextrinase (LLCD) gene was cloned from the genome of Lactococcus lactis subsp. lactis KCTC 3769 (ATCC 19435), which encodes 584 amino acids with the predicted molecular mass of 68.7 kDa. KCTC 3769 shares approximately 40% of amino acid sequence identity with the CDase-family of enzymes. The dimeric enzyme with C-terminal six-histidines was heterologously expressed and purified from recombinant E. coli. LLCD showed the highest activity against ${\beta}$-cyclodextrin (CD) at pH 7.0 and $37^{\circ}C$. In particular, LLCD exhibited extremely low activity against starch and pullulan, while its CD-hydrolyzing activity was about 80 times higher than starch. Due to its much higher activity on CD over starch, LLCD has been identified as a member of CDases. However, LLCD can be distinguished from the other common CDases on the basis of its extremely low hydrolyzing activity against starch, pullulan, and acarbose.

• Journal of Sericultural and Entomological Science
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• v.48 no.1
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• pp.11-15
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• 2006

Antheraea yamamai silk fibroin powder was prepared by treatment with HCl. The prepared A. yamamai fibroin hydrolysate was characterized by gel filtration chromatography, amino acid analysis, X-ray diffractometry, circular dichroism analysis, differential thermal analysis, and thermogravimetry. The average molecular weight of A. yamamai powder was about 430 and the major amino acids composed of the powder were Ala and Ser. According to XRD analysis, A. yamamai silk powder showed sharp diffraction peaks at $2{\theta}=20.34^{\circ}\;and\;31.5^{\circ}$. CD spectrum showed a peak around 220 nm and a should 215 nm, assigned to ${\alpha}-helix\;and\;{\beta}-sheet$ structure, respectively. DSC and TGA showed that the maximum degradation temperature of powder was around $250{\sim}270^{\circ}C$. Moreover, no harmful heavy metal was contained in the A. yamamai silk fibroin hydrolysate.

• The Journal of Korean Medicine
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• v.26 no.3 s.63
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• pp.27-42
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• 2005

Objectives : Alzheimer's disease (AD) is a progressive and fatal neurodegenerative disease characterized by amyloid plaques and neurofibrillary tangles. These plaques are associated with degenerating neuronal processes and consist primarily of fibrillary aggregates of beta-amyloid$ protein, generated from amyloid precursor protein (APP). Another amyloidogenic fragment, the carboxyl terminus (CT) of APP, which is composed of 99-105 amino acid residues containing the complete $A{\beta}$ sequence, also appears to be toxic to neurones. Recent evidence suggest that CT105, carboxy terminal 105 amino acids peptide fragment of APP, may be an important factor causing neurotoxicity in AD. Methods : Although a variety of oriental prescriptions including Pinelliae rhizoma have traditionally been utilized for the treatment of AD, their pharmacological effects and action mechanisms have not yet been fully elucidated. In the present study, we investigated effects of the dichloromethane extract of Pinelliae rhizoma (PINR) on neurotoxicity and the formation of reactive oxygen species (ROS) and nitric oxide (NO) in SK-N-SH cells overexpressed with CT105. In addition, we evaluated its radical scavenging activity and effects on acetylcholinesterase (AChE) activity. Furthermore, effects on cognitive deficits induced by scopolamine treatment in rats were evaluated. Results ; We found in this study that PINR significantly inhibited apoptotic neuronal death induced by CT105 overexpression in SK-N-SH cells. Based on morphological examinations by phase-contrast microscopy, PINR reversed apoptotic changes of CT105-expressed cells. It was also found that PINR significantly promoted neurite outgrowth and inhibited formation of ROS nd NO. PINR was shown to scavenge DPPH radicals and noncompetitively inhibit AChE activity. Furthermore, it reduced scopolamine-induced memory impairment in rata, assessed by passive avoidance test. Conclusions : Taken together, these results demonstrate that PINR exhibits neuroprotective, antioxidant, and memory enhancing effects, and therefore may bs beneficial for the treatment of AD.