• Title/Summary/Keyword: biochemical activity

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Water Extract of Hovenia dulcis Suppressed Lipid Peroxidation and Improved Renal Function in $CCl_4$ Intoxicated Rats ($CCl_4$를 투여한 랫드에서 헛개나무 열수추출물의 지질과산화 억제와 신기능 개선 작용)

  • park Yeun Woo;Yang Si Yang;Lee Min Kyung;Jin Ju Young;Cho Jung Hee;Kim Ki Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.868-873
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    • 2004
  • Renal dysfunction could be developed as the secondary disease of liver cirrhosis. Delayed or suppresed lipid peroxidation by the treatment with physiological active substances could be explained as the antioxidative and protective effect in tissue damage. In this study, we investigated an antioxidative effect and renal function improvement of Hovenia dulcis in liver fibrosis(cirrhosis) induced rats. The female Sprague-Dawley rats (180∼210 g) were divided into 3 groups (Normal, AC: CCl₄ mixture treated group, AC-HV: CCl₄ mixture+ Hovenia dulcis treated group) and renal damage was developed by CCl₄ mixture administration in 4 weeks (0.8 ㎖/rat). The tissue of kidney and liver and sera were used for quantitative measurement of enzyme activity, MDA and Hyp. The histological change and gene expression of collagen α1(III) mRNA and a1(IV) mRNA were observed by Masson's trichrome staining and RT-PCR. As a result, the clinical biochemical parameters of liver function (AST and ALT) in sera of AC-HV group showed significantly 46.4% and 104.8% lower (p<0.005), and the level of ALP and BUN as the parameter of protein urine and azotemia showed 17.8 % and 25.8 % lower than in AC group. In AC-HV group, the concentration of MDA in kidney and liver was decreased significantly 15.8% and 21.3% when compared with AC group (p<0.01 -0.005). The content of Hyp in kidney of AC-HV group is merely higher than in AC group, in contrast to liver tissue. The expression of collagen α1(III) mRNA and collagen α1(IV) mRNA was decreased in AC, but both of collagen mRNA in normal and AC-HV group expressed fast similar. More massive lipid droplets, thicker collagen fiber bundles in portal triads and more formation of portal central septum were observed in the liver of AC group than in AC-HV group. In conclusion, CCl₄ mixture intoxication could be developed not only liver fibrosis(cirrhosis) but also renal dysfunction by the massive lipid peroxidation and suppression of interstitial collagen and basement membrane collagen synthesis. And the water extract of Hovenia dulcis may be possessed the antioxidative and protective effect and improvement of kidney function in renal dysfunction induced rats.

Production and Characterization of α-Galactosidases from Two Bacillus licheniformis Isolates (Bacillus licheniformis 분리균 2종의 α-Galactosidase 생산성과 효소특성)

  • Jin, Hyun Kyung;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.195-203
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    • 2015
  • Two bacterial strains, Bacillus licheniformis YB-1413 and YB-1414, producing extracellular α-galactosidase, were obtained from homemade Doenjang. On the basis of their biochemical properties, 16S rRNA sequences and random amplified polymorphic DNA patterns by polymerase chain reaction, they were found to be somewhat different from one another. α-Galactosidase productivities of the two isolates were increased by wheat bran, but drastically decreased by melibiose, raffinose and sucrose which were used as carbon sources. The enzyme productivities were increased by yeast extract as a nitrogen source with maximum levels of 1.87 U/ml for YB-1413 and 1.69 U/ml for YB-1414, respectively. The enzymes of both isolates exhibited maximum activity for hydrolysis of para-nitrophenyl-α-D-galactopyranoside (pNP-αGal) under reaction conditions of pH 6.0 and 45℃. Their hydrolyzing activities for pNP-αGal were drastically decreased by the addition of low concentrations of ribose and galactose. They were capable of hydrolyzing completely α-1,6 linked galactosyl residue in melibiose, raffinose and stachyose, which are known to be anti-nutritional factors in products of soybean and legume. In relation to the latter, the isolates YB-1413 and YB-1414 have potential applicability in improving soybean-fermented foods and the nutritional value of soybean feed.

Gene Cloning and Expression of Trehalose Synthase from Thermus thermophilus HJ6 (Thermus thermophilus HJ6 유래 내열성 Trehalose Synthase의 유전자 클로닝 및 발현)

  • Kim, Hyun-Jung;Kim, Han-Woo;Jeon, Sung-Jong
    • Microbiology and Biotechnology Letters
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    • v.36 no.3
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    • pp.182-188
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    • 2008
  • A hyperthermophilic bacteria (strain HJ6) was isolated from a hot springs located in the Arima-cho, Hyogo, Japan. The cells were long-rod type ($2-4{\mu}m$), about $0.4{\mu}m$ in diameter. The pH and temperature for optimal growth were 6.5 and $80^{\circ}C$, respectively. Phylogenetic analysis based on the 16S rDNA sequence and biochemical studies indicated that HJ6 belonged to the genus Thermus thermophilus (Tt). The gene encoding the Trehalose synthase (TS) was cloned and sequenced. The open reading frame (ORF) of the TtTS gene was composed of 2,898 nucleotides and encoded a protein (975 amino acids) with a predicted molecular weight of 110.56 kDa. The deduced amino acid sequence of TtTS showed 99% and 83% identities to the Thermus caldophilus TS and Meiothermus ruber TS, respectively. TtTS gene was expressed in Escherichia coli cells, and the recombinant protein was purified to homogeneity. The optimal temperature and pH for Trehalose synthase activity were found to be $80^{\circ}C$ and 7.5, respectively. The half-life of heat inactivation was about 40 min at $90^{\circ}C$. The maximum trehalose conversion rate of maltose into trehalose by the enzyme increased as the substrate concentration increased, and reached 55.7% at the maltose concentration of 500 mM, implying that the enzyme conversion was dependent of the substrate concentration.

Biochemical Characteristics of Whole Soybean Cereals Fermented with Aspergillus Strains. (Aspergillus속 균류들을 이용한 콩알메주 발효의 생화학적 특성)

  • Kim, Dong-Ho;Kim, Seung-Ho;Choi, Nak-Sik;Bae, Seok;Jeon, Soon-Bae
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.551-557
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    • 1998
  • Whole soybean cereal was fermented with four Aspergillus strains in pilot meju fermentation system. The pH range of the product was 7.40~7.98, the contents of reducing sugar and amino-nitrogen were 0.04~2.78%, 178~309 mg%, respectively and that of free fatty acid ranged 2.67~5.05%. The components of the amino acid, organic acid, free sugars and fatty acid showed distinctive patterns among four groups of fermented soybean cereals. Amylase activity and carbohydrate degradation rate of A. usami was higher than other strains. But protease and protein degradation rate, lipase and lipid degradation rate were similar in four strains. The odor concentrates of soybean cereals fermented with Aspergillus strains were different from Bacillus strains. Especially, pyrazine components, the main and common flavor chemicals in Bacillus strains, were not determined in this study and Aspergillus specific components were 9-methyl-acridine, dl-limonene and 2,3-butanediol. Soybean paste, made from A. oryzae fermented soybean cereal, showed excellent sensory evaluation.

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Studies on Isolation of Pasteurella multocida from Slaughtered Pigs, Microbiological and Serological Properties, Antibiotic Susceptibility of the Isolates and Pathological Observation of Their Lungs (도축돈에서 Pasteurella multocida의 분리, 분리주에 대한 미생물학적, 혈청학적성상 및 항생물질의 감수성조사와 돈폐에 대한 병리학적 관찰에 관하여)

  • Lee Hak-Cheol;Ham Tai-Soo;Chung Yoo-Yeol;Cho Seong-Lyong;Lee Jag-Hyeon
    • Journal of Veterinary Clinics
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    • v.6 no.2
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    • pp.307-318
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    • 1989
  • In recent years much attention has been paid to swine respiratory infection caused by Pasteurella(P) multocida with rapid expansion of pork Industry in Korea. The present study was performed to observe the etiologic situation of P. multocida infection by bacteriological, serological(serotyping) and pathological examinations with the lungs respectively. In addition antibiotic susceptibility test was carried out against the isolated strains of P. multocida. The results obtained are as follows : 1. Eighteen strains(12.8%) wert isolated from the 140 cases of swine lungs examined, and biological and biochemical characteristics of the isolates were the sam as those in the references of other workers, whereas some differences were observed in sugar fermentation and enzyme activity according to the strain of isolates. 2. Capsular serotyping performed on 18 P. multocida revealed that 13 strains(72.2%) were A type and 5 strains(27.8%) were D type, respectively. 3. When serotyping was performed against somatic antigen on 18 strains capsular types of which were identified as described above 9(50%), 3(16.7%) and 4(22.2%) strains belong to 1:A, 3:A and 2:D, respectively, but untypable 2 strains(11.1%) were observed. 4. Antibiotic susceptibility test by employing disc method for 24 kinds of drugs revealed that 15 kinds of antibiotics were sensitive to 18 strains of P. multocida isolated such as ampicillin(l00%), penicillin(100%), cloxacillin(56%), piperacillin(70%), cefotaxime(30%), minocycline(60%), chloramphenicol(95%), erythromycin(39%), kanamycin(17%), gentamicin(70%), amikasin(30%), colistin(78%) and nalidixic acid(5%), respectively, but resistant to 9 kinds of antibiotics such as sulpenicillin, cefazolin, cephalothin, cefametazol, cefoperazone, kitasamycin, oleandomycin. lincomycin and bacitracin. 5. Pathological features of 60 cases of swine lungs indicated that pneumonic .lesions were observed in 38 cases(63.3%) examined by macroscopic finding, in which lesions of 8 cases(13.4%) would correspond to those of mycoplasmal infection, and 30 cases(50%) were similar to viral infection by histopathological finding, whereas 22 cases(36.7%) were considered to be normal by ecropsy or histopathological finding.

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Screening of Bacteria Producing Lipase from Insect Gut: Isolation and Characterization of a Strain, Burkholderia sp. HY-10 Producing Lipase (곤충 장내미생물로부터 lipase 생산능력이 우수한 Burkholderia sp. HY-10 균주의 분리 및 특성)

  • Park, Doo-Sang;Oh, Hyun-Woo;Bae, Kyung-Sook;Kim, Hyang-Mi;Heo, Sun-Yeon;Kim, Nam-Jung;Seol, Kwang-Youl;Park, Ho-Yong
    • Korean journal of applied entomology
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    • v.46 no.1 s.145
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    • pp.131-139
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    • 2007
  • From the course of screening of useful enzyme producing microorganism from insect guts, we isolated 9 lipase producing strains and their lipase producing activities were tested. 16S rDNA sequence analysis showed that they were Gram negative bacteria grouped on Serratia sp., Pseudomonas sp., and Burkholderia sp.. Among them, an excellent lipase producing strain, Burkholderia sp. HY-10 identified by 16S rDNA analysis and biochemical methods, was further studied its lipase producing characteristics. It was isolated from a longcorm beetle, Prionus insularis and showed cell density dependent lipase producing activity in the culture media that contained olive oil as a carbon source. Maximum lipase production was achieved in the M9 media containing 0.5% yeast extract and 0.5% olive oil when cultured at $30^{\circ}C$ for 36-42 hrs.

Immunocytochemical Localization of Metallothionein in Gastric Adenocarcinoma (위암 조직내 Metallothionein의 면역 세포화학적 연구)

  • Yang, Seung-Ha;Shin, Kil-Sang;Kim, Wan-Jong
    • Applied Microscopy
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    • v.32 no.4
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    • pp.411-419
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    • 2002
  • Metallothionein (MT) is a family of ubiquitous, low molecular weight ($6,000{\sim}7,000D$), cysteine-rich ($30{\sim}35%$) inducible protein with a high affinity to metal ions and has no aromatic amino acids and histidine. Some of the known functions of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Also, this protein has been reported to involve in tumor pathophysiology and therapy resistance. MT expression may affect a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. Many reports on the physiological and biochemical properties of MT have been published, but ultrastructural reports on the localization of MT in human gastric cancer tissues are extremely rare. The present study was undertaken to examine the ultrastructural features and the localization of MT within the gastric adenocarcinoma. Ultrastructures of gastric cancer cells were characterized by the high nuclear cytoplasmic ratio, the interdigitation between cells, the irregular nucleus containing much heterochromatin and the wide distribution of free ribosomes in the cytoplasm. Immunohistochemical reaction for MT was prominent in the gastric adenocarcinoma. And the immunogold labellings were more prominent within the nucleus than the cytoplasm. Particularly, immunogold particles were numerously seen at nulcleolus or nucleolar associated heterochromatin. These results suggest that MT expression by gastric cancer cells is associated with cell proliferative activity and is possibly synthesized in the cytoplasm, and then the protein is transported into the nucleus to participate in any transcriptional steps.

Biochemical characterization of a novel extracellular pullulan 6-glucanohydrolase from Bacillus circulans S-1

  • Lee, Moon-Jo;Park, Cheon;Park, Joon-Ho;Chung, Kang-Hyun;Nam, Kyung-Soo;Park, Jin-Woo;Kim, Cheorl-Ho
    • Journal of Life Science
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    • v.10 no.3
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    • pp.307-316
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    • 2000
  • Bacillus circulans S-1 extracellular pullulan 6-glucanohydrolase (EP) (EC 3.2.1.41) has been characterized with a purified enzyme of 140 kDa. The N-terminal amino acid sequence of the purified enzyme was P-L-N-M-S-Q-P. The enzyme displayed a temperature optimum of around $60^{\circ}C$ and a pH optimum of around pH 9.0. The enzyme was stable to incubation from pH 4.0 to pH 11.0 at $4^{\circ}C$ for 48h. The presence of substrate allowed the protection of the enzyme from heat inactivation. The activity of the enzyme was stimulated by several metal ions such as Mn2+ and Ca2+. The enzyme had an apparent Km of 7.92 mg/ml for pullulan. The purfied enzyme completely hydrolysed pullulan to maltotriose.

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Safety Test of Brown Rice Expressing Arabidopsis Calcium Transporter by Feeding Trial in Mice (애기장대 칼슘수송체를 발현하는 형질전환 현미의 생쥐 식이를 통한 안전성 평가)

  • Kim, Kyung-Min;Kim, Chang-Kil;Kim, Byung-Oh
    • Journal of Life Science
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    • v.18 no.10
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    • pp.1390-1394
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    • 2008
  • Previously, we found that the transgenic rice plants over-expressing the Arabidopsis $H^+/Ca^{2+}$ antiporter CAX 1 (accession no. U57411) gene accumulated 2.7 to 7.5-fold more calcium in the T3 rice grains as compared to those of control. To examine physiological safety of the $T_3$ rice grains, the effect of the $T_3$ brown rice on change in levels of body weight and white blood cells was compared with that of the control Ilpum brown rice by feeding trial in mice. During the feeding trial for one month, there was no significant difference between two mice groups, which were fed by the $T_3$ brown rice or Ilpum brown rice. There were no detectable differences in their effects on immune functions including plaque-forming unit, peritoneal macrophage number, and NK-cell activity. In addition, biochemical analysis of the blood failed to exhibit any difference between two mice groups. Together, these results suggested that the $T_3$ brown rice, which was produced from a genetically modified organism (GMO), might be safe and possess a potential to be applicable as calcium-fortified feed or food. Long-term safety of the $T_3$ brown rice, however, remains to be elucidated.

Effect of Silkworm (Bombyx mori) Excrement Powder on the Alcoholic Hepatotoxicit in Rats (알코올성 간독성에 미치는 누에배설물의 영향)

  • Kim, Yong-Soon;Kim, Kee-Young;Kang, Pil-Don;Cha, Jae-Young;Heo, Jin-Sun;Park, Bo-Kyung;Cho, Young-Su
    • Journal of Life Science
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    • v.18 no.10
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    • pp.1342-1347
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    • 2008
  • The purpose of present study was to investigate the protective effect of silkworm excrement powder (SEP) on alcohol-induced hepatotoxicity in rats. Semisynthetic diet supplemented with SEP (3%, w/w) given to alcohol-feeding rats for 30 days, then blood and tissues were collected, processed and used for alcohol concentration mensuration, various biochemical estimations and histopathological examination. Chronic alcohol administration resulted in significantly increase in the activities of the clinically important liver marker enzymes, alanine aminotransferase (ALT), aspartate aminotransferase (AST), $\gamma$-glutamyl transpeptidase ($\gamma$-GTP) and lactate dehydrogenase (LDH). Also, a highly significant increase in the blood alcohol level by alcohol treatment was observed. But alcohol-induced elevation of ALT and LDH levels markedly prevented and the level of blood alcohol decreased in SEP treated rats as compared to alcohol-administered control rats. SEP supplementation showed highly decreased the concentrations of total lipid, triglyceride and cholesterol in serum, as compared with alcohol treated control rats. Alcohol treatment induced the marked accumulation of large lipid droplets, hepatocytes necrosis and inflammation in the liver, but SEP administration attenuated to alcohol-induced accumulation of lipid droplets and hepatocyte necrosis. The results indicated that SEP may exert a protective effect against alcoholic hepatotoxicity through decreasing the activity of hepatic marker enzymes.