• Korean Journal of Microbiology
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• v.51 no.4
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• pp.319-328
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• 2015

The $sdu1^+$ gene encodes Sdu1, a PPPDE superfamily member of deubiquitinating enzymes (DUBs) in Schizosaccharomyces pombe. Sdu1 was previously shown to contain an actual ubiquitin C-terminal hydrolase (UCH) activity using the recombinant plasmid pYSTP which harbors the $sdu1^+$ gene. This work was designed to assess a thermotolerant role of Sdu1 against high incubation temperatures. In the temperature-shift experiments, the S. pombe cells harboring pYSTP grew much better after the shifts to $37^{\circ}C$ and $42^{\circ}C$, when compared with the vector control cells. After being shifted to $37^{\circ}C$ and $42^{\circ}C$ for 6 h, the S. pombe cells harboring pYSTP contained lower reactive oxygen species (ROS) levels, compared with the vector control cells. The nitric oxide (NO) levels of the S. pombe cells harboring pYSTP were slightly lower than those of the vector control cells in the absence or presence of the temperature shifting. The total glutathione (GSH) levels of the S. pombe cells harboring pYSTP were significantly higher than those of the vector control cells. Total superoxide dismutase (SOD) and GSH peroxidase activities were also higher in the S. pombe cells harboring pYSTP after the temperature shifts than in the vector control cells. In brief, the S. pombe Sdu1 plays a thermotolerant role against high incubation temperature through the down-regulation of ROS and NO and the up-regulation of total GSH content, total SOD and GSH peroxidase activities.

• The Journal of Korean Medicine
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• v.16 no.2 s.30
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• pp.320-336
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• 1995

SANGNYANGHYOLTANG(SYT) is one of the most important prescription that has been used in oriental medicine for diabetes mellitus. The sudy was done in order to elucidate the anti-diabetic effect of SYT. After pretreatment of SYT(1,000mg／kg) for 6 weeks, the effect of of SYT was prevented on serum liver function test and hepatic lipid peroxide content in rats i.v. injected with streptozotocin(STZ, 50mg／kg, tail vein) 5 weeks after pretreatment of SYT. The hepatic microsomal cytochrome P-450 and aniline hydroxylase were significantly decreased, and aminopyrine N-demethylase activity was significantly increased in SYT-STZ group as compared with control group. Changes in aldehyde oxidase, xanthine oxidase, superoxide dismutase, catalase, epoxide hydrolase, UDP-glucuronyltransferase and sulfotransferase activities were not significantly different in any of the group. The cytosolic glutathione S-transferase activity was significantly decreased in SYT-STZ group as compared with control group. The selenium-independent glutathione peroxidase was significantly increased in SYT-STZ group as compared with control group, but there was no significant difference in selenium-dependent glutathione peroxidase in any of the groups. The hepatic glutathione concentration was significantly increased in SYT-STZ group as compared with control group, and ${\gamma}-glutamylcystein$ synthetase and glutathione reductase activities were not significantly different in any of the groups. The hepatic lipid peroxide content, serum aminotransferase and sorbitol dehydrogenase activities were slightly decreased in significantly in SYT-STZ groups.

• Tuberculosis and Respiratory Diseases
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• v.55 no.1
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• pp.88-97
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• 2003

Background : Although smoking is a major cause of chronic obstructive pulmonary disease (COPD), only 10-20% of cigarette smokers develop symptomatic COPD, which suggests the presence of genetic susceptibility. This genetic susceptibility to COPD might depend on variations in the activities of the enzyme that detoxify hazardous chemical products, such as microsomal epoxide hydrolase (mEPHX) and glutathione-S transferase M1 subunit (GSTM1) genes. Methods : The genotypes of 58 patients with COPD, and 79 age matched control subjects, were determined by a polymerase chain reaction, followed by restriction fragment length polymorphism (PCR-RFLP) for the mEPHX, and multiplex PCR for the GSTM1. Results : GSTM1 was deleted in 53.3% of the subjects. There was no difference in GSTM1 deletion rates between the COPD patients (32/58, 55.2%) and the control subjects (41/79, 51.9%). The combination patterns of two polymorphisms of mEPHX showed slow enzyme activity in 29(21.2%), normal in 73(53.3%) and fast in 32(23.4%). The COPD group (7/57, 12.3%) showed a significantly lower incidence of slow enzyme activity compared to the control subjects (22/77, 28.6%, p<0.05). However, when the COPD and control groups were compared with smokers only, there were no significant differences in the genotypes of GSTM1 and mEPHX. Conclusion : The genotypes of GSTM1 and mEPHX were not significant risk factors of COPD in this cohort of study.

• Korean Journal of Poultry Science
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• v.51 no.2
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• pp.107-116
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• 2024

The study was conducted to examine age-related development of digestive organs, intestinal microvilli hydrolase, and hepatic antioxidant enzyme in Korean native chicks (KNC) aged from 0-d to 28-d of post-hatching. Body weight did not significantly increase from 0-d to 3-d-old, but after that remarkably increased from 3-d to 28-d-old (P<0.05). The relative weight (g/100 g of BW) of the proventriculus, gizzard, and liver was significantly higher at 3- and 7-d-old chicks than that of the other ages. The relative weight of the intestine, mucosal tissues, and pancreas was markedly developed at the ages of 3-, 14-, and (or) 21-d-old chicks (P<0.05). In the small intestine, the specific activities of maltase and sucrose were significantly higher at 14-d-old compared with the other ages (P<0.05). Leucine aminopeptidase activity showed a constant level from 0- to 28-d-old without significance. The specific activity of alkaline phosphatase was significantly higher at 0-d-old compared with the other ages (P<0.05). In the liver, the specific activities of superoxide dismutase, glutathione peroxidase, and glutathione S-transferase were shown to be lowest at 0-d-old, but they continued to increase as the age increased. The lipid peroxidation was significantly high at the age of 21-d (P<0.05), after that its level decreased at 28-d old. In conclusion, the KNC rapidly developed digestive organs and intestinal microvilli hydrolase activity from 3- to 14-d-old after hatching. Hepatic antioxidant enzyme activity continued to increase as the age increased after hatching, resulting in 28-d-old chicks showing the highest antioxidant enzyme activity in the KNC.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.402-406
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• 2005

Lipomyces starkeyi produces a novel glucanhydrolase containing endo-dextranase and ${\alpha}-amylase$ activities. A cDNA from L. starkeyi encoding a dextranase was isolated and characterized. The 2,052 kb cDNA fragment (lsd1) carrying dextranase gene showed one open reading frame (ORF) composed of 1,824 bp flanked by a 41 bp 5'-UTR and a 184 bp 3'-UTR including a poly(A) tail of 27 bp. The ORF encodes for a 608 amino acid with a predicted molecular mass of 67.6 kDa. There was 77% deduced amino acid sequence identity between the LSD1 dextranase and the dextranase from Penicillium minioluteum. The primary structure of the dextranase from L. starkeyi has distant similarity with enzymes belonging to glycosyl hydrolase family 49. The lsd1 protein was expressed in the Saccharmyces cerevisiae under control of GAL1 promoter and active dextranase was produced.

• Journal of Microbiology and Biotechnology
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• v.27 no.3
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• pp.542-551
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• 2017

Small phytochemicals have been successfully adopted as antibacterial chemotherapies and are being increasingly viewed as potential antibiofilm agents. Some of these molecules are known to repress biofilm and toxin production by certain bacterial and yeast pathogens, but information is lacking with regard to the genes allied with biofilm formation. The present study was performed to investigate the inhibitory effect of burdock root extract (BRE) and of chlorogenic acid (CGA; a component of BRE) on clinical isolates of Klebsiella pneumoniae. BRE and CGA exhibited significant antibiofilm activity against K. pneumoniae without inflicting any harm to its planktonic counterparts. In vitro assays supported the ${\beta}$-lactamase inhibitory effect of CGA and BRE while in silico docking showed that CGA bound strongly with the active sites of sulfhydryl-variable-1 ${\beta}$-lactamase. Furthermore, the mRNA transcript levels of two biofilm-associated genes (type 3 fimbriae mrkD and trehalose-6-phosphate hydrolase treC) were significantly downregulated in CGA- and BRE-treated samples. In addition, CGA inhibited biofilm formation by Escherichia coli and Candida albicans without affecting their planktonic cell growth. These findings show that BRE and its component CGA have potential use in antibiofilm strategies against persistent K. pneumoniae infections.

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1699-1706
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• 2014

A lichenase gene (mt-lic) was identified for the first time through function-based screening of a soil metagenomic library. Its deduced amino acid sequence exhibited a high degree of homology with endo-${\beta}$-1,3-1,4-glucanase (having both lichenase and chitosanase activities), encoded by the bgc gene of Bacillus circulans WL-12. The recombinant lichenase overexpressed and purified from Escherichia coli was able to efficiently hydrolyze both barley ${\beta}$-glucan and lichenan. The enzyme showed maximal activity at a pH of 6.0 at $50^{\circ}C$, with Azo-barley-glucan as the substrate. The metal ions $Mn^{2+}$, $Mg^{2+}$, $Ca^{2+}$, and $Fe^{2+}$ enhanced the enzymatic activity, whereas the $Cu^{2+}$ and $Zn^{2+}$ ions inhibited the enzymatic activity. The $K_m$ and $V_{max}$ values of the purified lichenase were determined to be 0.45 mg/ml and 24.83 U/min/mg of protein, respectively.

• Microbiology and Biotechnology Letters
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• v.48 no.3
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• pp.276-286
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• 2020

Probiotics are live microorganisms that, when administered in adequate amounts, confer health benefits to the host. This study was conducted for the isolation of potential lactic acid bacteria (LAB) with probiotic properties from goat milk and yogurt. Several tests were conducted in vitro using the standard procedures for evaluating the inhibitory spectra of LAB against pathogenic bacteria; tolerance to NaCl, bile salt, and phenol; hemolytic, milk coagulation, and bile salt hydrolase activities; gastrointestinal transit tolerance; adhesion properties; and antibiotic susceptibility. Among 40 LAB strains screened according to culture characteristics, five isolates exhibited antagonistic properties. Three were identified as Pediococcus acidilactici, and two were identified as Enterococcus faecium, exploiting 16S rRNA gene sequencing. All the isolates succeeded in the gastrointestinal transit tolerance assay and successively colonized mucosal epithelial cells. Based on the results of these in vitro assays, both P. acidilactici and E. faecium can be considered as potential probiotic candidates.

• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1711-1716
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• 2010

A gene encoding the ${\beta}$-xylosidase/${\alpha}$-arabinofuranosidase (XylC) of Paenibacillus woosongensis was cloned into Escherichia coli. This xylC gene consisted of 1,425 nucleotides, encoding a polypeptide of 474 amino acid residues. The deduced amino acid sequence exhibited an 80% similarity with those of both Clostridium stercorarium ${\beta}$-xylosidase/${\alpha}$-N-arabinosidase and Bacillus cellulosilyticus ${\alpha}$-arabinofuranosidase, belonging to the glycosyl hydrolase family 43. The structural gene was subcloned with a C-terminal His-tag into a pET23a(+) expression vector. The His-tagged XylC, purified from a cell-free extract of a recombinant E. coli BL21(DE3) Codon Plus carrying a xylC gene by affinity chromatography, was active on para-nitrophenyl-${\alpha}$-arabinofuranoside (pNPA) as well as para-nitrophenyl-${\beta}$-xylopyranoside (pNPX). However, the enzymatic activities for the substrates were somewhat incongruously influenced by reaction pHs and temperatures. The enzyme was also affected by various chemicals at different levels. SDS (5 mM) inhibited the enzymatic activity for pNPX, while enhancing the enzymatic activity for pNPA. Enzyme activity was also found to be inhibited by addition of pentose or hexose. The Michaelis constant and maximum velocity of the purified enzyme were determined for hydrolysis of pNPX and pNPA, respectively.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.800-805
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• 2007

Two endoglucanases with processive cellulase activities, produced from Fomitopsis palustris grown on 2% microcrystalline cellulose(Avicel), were purified to homogeneity by anion-exchange and gel filtration column chromatography systems. SDS-PAGE analysis indicated that the molecular masses of the purified enzymes were 47 kDa and 35 kDa, respectively. The amino acid sequence analysis of the 47-kDa protein(EG47) showed a sequence similarity with fungal glycoside hydrolase family 5 endoglucanase from the white-rot fungus Phanerochaete chrysosporium. N-terminal and internal amino acid sequences of the 35-kDa protein(EG35), however, had no homology with any other glycosylhydrolases, although the enzyme had high specific activity against carboxymethyl cellulose, which is a typical substrate for endoglucanases. The initial rate of Avicel hydrolysis by EG35 was relatively fast for 48 h, and the amount of soluble reducing sugar released after 96 h was $100{\mu}g/ml$. Although EG47 also hydrolyzed Avicel, the hydrolysis rate was lower than that of EG35. Thin layer chromatography analysis of the hydrolysis products released from Avicel indicated that the main product was cellobiose, suggesting that the brown-rot fungus possesses processive EGs capable of degrading crystalline cellulose.