• BMB Reports
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• v.34 no.1
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• pp.39-42
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• 2001

Previous studies indicate that B. subtilis protoporphyrinogen oxidase is poorly inhibited by diphenyl ether herbicides. To better understand the basis of this insensitivity, the enzyme was overexpressed as a soluble protein in E. coli, purified and characterized. The mechanism of oxidation of B. subtilis protoporphyrinogen IX was studied and the enzyme kinetic parameters were determined for protoporpyrinogen IX; $K_m$, and $k_{cat}$ were $6.3\;{\mu}M$ and $0.028\;h-^1$, respectively. The enzyme required flavin adenine dinucleotide as a cofactor and its activity was enhanced by 1 mM n-octylglucopyranoside. The nonenzymatic oxidation rate was dependent on the concentration of protoporphyrinogen IX, suggesting that the reaction involves a pre-equilibrium step followed by a rate-limiting step.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.112-114
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• 1998

For enhancement of cis,cis-muconate productivity from benzoate, catechol 1,2-dioxygenase (C12O) which catalyzes the rate-limiting step (catechol conversion to cis,cis-muconate) was cloned and expressed in recombinant Pseudomonas putida BCM114. At higher benzoate concentrations (more than 15 mM), cis,cis-muconate productivity gradually decreased and unconverted catechol was accumulated up to 10 mM in the cae of wild-type P. putida BM014, whereas cis,cis-muconate productivity continuously increased and catechol was completely transformed to cis,cis-muconate for P. putida BCM114. Specific C12O activity of P. putida BCM114 was about three times higher than that of P. putida BM014, and productivity was enhanced more than two times.

• Journal of Forest and Environmental Science
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• v.26 no.3
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• pp.171-179
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• 2010

Effect of chromium (Cr) stress on antioxidant enzyme activities and malondialdehyde (MDA) content were investigated in leaves and roots of mangrove (italic (L.) Druce) seedlings. Cr toxicity effects were also assessed on young seedlings. The seedlings were grown in green house condition for three months in nutrient solution with 0, 0.5, 1, 1.5, 2, 2.5, and 3 mg $L^{-1}$ $CrCl_3$. This study showed that Cr led to the change of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) and activities at different concentrations. The activity of antioxidant enzymes in leaves of K. candel seedlings indicates that enzymes engaged in antioxidant defense in certain level especially in low concentration of Cr treatments. The activities of SOD and POD were activated by Cr in the root level, while CAT activity was inhibited. CAT activity decreased in response to high concentrations of Cr. In the present study indicated that SOD in root was active in scavenging the superoxide produced by Cr. Both in roots and leaves, an increase in malondialdehyde (MDA) content was observed with increase in metal concentration and exposure periods. Our finding indicated that the high concentration of excessive Cr supply may interfere with several metabolic processes of seedlings, causing toxicity to plants as exhibited by chlorosis, necrosis, photosynthetic impairing and finally, plant death.

• BMB Reports
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• v.32 no.5
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• pp.486-491
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• 1999

An engineered cDNA from Phanerochaete chrysosporium encoding both the mature and propeptide-sequence regions of lignin peroxidase H2 (Lip H2) was overexpressed in Escherichia coli BL21 (DE3) to evaluate its catalytic characteristics and potential application as a pollution scavenger. All expressed proteins were aggregated in an inactive inclusion body, which might be due to inherent disulfide bonds. Active enzyme was obtained by refolding with glutathione-mediated oxidation in refolding solution containing $Ca^{2+}$, heme, and urea. Propeptide-sequence region was not processed as evidenced by N-terminal sequence analysis. Recombinant Lip H2 (rLip H2) had the same physical properties of the native protein but differed in the $K_{cat}$. Catalytic efficiency ($k_{cat}/K_m$) of rLip H2 was slightly higher than that of the native enzyme. In order to express an active protein, fusion systems with thioredoxin or Dsb A, which have disulfide isomerase activity, were used. The fused proteins expressed by the Dsb A fusion vector were aggregated, whereas half of the thioredoxin fusion proteins were recovered as a soluble form but still catalytically inactive. These results suggest that Lip H2 may not be expressed as an active enzyme in Escherichia coli although the activity can be recovered by in vitro refolding.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1122-1132
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• 2018

In this study, we attempted to find new and efficient microbial enzymes for producing rare sugars. A ribose-5-phosphate isomerase B (OsRpiB) was cloned, overexpressed, and preliminarily purified successfully from a newly screened Ochrobactrum sp. CSL1, which could catalyze the isomerization reaction of rare sugars. A study of its substrate specificity showed that the cloned isomerase (OsRpiB) could effectively catalyze the conversion of $\text\tiny{L}$-rhamnose to $\text\tiny{L}$-rhamnulose, which was unconventional for RpiB. The optimal reaction conditions ($50^{\circ}C$, pH 8.0, and 1 mM $Ca^{2+}$) were obtained to maximize the potential of OsRpiB in preparing $\text\tiny{L}$-rhamnulose. The catalytic properties of OsRpiB, including $K_m$, $k_{cat}$, and catalytic efficiency ($k_{cat}/K_m$), were determined as 43.47 mM, $129.4sec^{-1}$, and 2.98 mM/sec. The highest conversion rate of $\text\tiny{L}$-rhamnose under the optimized conditions by OsRpiB could reach 26% after 4.5 h. To the best of our knowledge, this is the first successful attempt of the novel biotransformation of $\text\tiny{L}$-rhamnose to $\text\tiny{L}$-rhamnulose by OsRpiB biocatalysis.

• Tuberculosis and Respiratory Diseases
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• v.83 no.3
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• pp.228-233
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• 2020

Background: The Bronchiectasis Health Questionnaire (BHQ) is a simple and repeatable, self-reporting health status questionnaire for bronchiectasis. We have translated the original version of the BHQ into Korean using a standardized methodology. The purpose of this study was to assess the validity of the Korean version of the BHQ (K-BHQ) with Korean patients. Methods: Stable state patients with bronchiectasis from two academic hospitals were enrolled in this study. The validity was assessed by investigating the relationship between the K-BHQ scores and the Korean version of the Chronic Obstructive Pulmonary Disease Assessment Test (K-CAT) scores. We also investigated the relationship between the K-BHQ scores and other variables of the modified Medical Research Council's (mMRC) dyspnea scale, lung function, and exacerbations. Results: A total of 126 patients with bronchiectasis were enrolled. The mean age was 64.3 (standard deviation [SD], 9.7). Women comprised 53.2% of the patients. The mean forced expiratory volume in one second (FEV₁) was 60% of the predicted value (SD, 18.9%); the mean K-CAT score was 17.6 (SD, 9.1). The K-BHQ scores correlated strongly with the K-CAT scores (r=-0.656, p<0.001). There was significant correlation between the K-BHQ scores and the mMRC dyspnea scale (ρ=-0.409, p<0.001), FEV₁ (r=0.406, p<0.001), and number of exacerbations requiring hospitalization (ρ=-0.303, p=0.001). Conclusion: The K-BHQ is valid for assessing the health-related quality of life or health status of Korean bronchiectasis patients.

• Journal of Life Science
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• v.16 no.6
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• pp.984-988
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• 2006

Lipopolysaccharide(LPS) was posttreated after the 14 day-pretreatment of Ulva lactuca fractions(ULF), and their correlativity to enzymatic activity alteration was investigated in the liver of rats. ULF was intraperitoneally administered into rats at dose of $1m{\ell}/kg$ of 100 mg/kg concentration for 14 days. On the day 15, $1m{\ell}/kg$ of LPS was injected. The corelativity was examined by measuring the changed values of superoxide dismutase(SOD) in mitochondrial fraction and catalase(CAT), glutathione peroxidase(GPx) in liver homogenate. The results showed that LPS treatment decreased the high values of SOD, CAT, GPx to the low values, but ULF pretreatment increased the low values of SOD, CAT, GPx to the high values. It was suggested that ULF, LPS and antioxidative enzymes like SOD, CAT, GPx had the corelativity of the high-low-high pattern and that the ULF pretreatment played the proper preventive role in the protection against the LPS treatment-induced enzymatic inactivity in the water fraction.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.5
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• pp.588-597
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• 2010

We investigated the effect of quercetin on growth and plasma cholesterol level and the effects of quercetin pretreatment (Diet 1, 0%; Diet 2, 0.25%; and Diet 3, 0.5% quercetin) for 30 and 60 days on oxidative stress induced by hypo-osmotic conditions (17.5, 8.75, and 4 psu) in olive flounder. The weights of flounder were higher with Diet 3 than with Diet 1 and 2, which indicated that a high concentration (Diet 3) of quercetin was very effective in growth. Total cholesterol levels were lower with Diets 2 and 3 than with Diet 1, leading us to hypothesize that quercetin removed low-density lipoproteins from circulation and thereby reduced total cholesterol. To understand the antioxidant role of quercetin, we measured the mRNA expression and activities of superoxide dismutase (SOD) and catalase (CAT) and the $H_2O_2$ concentration in quercetin-treated flounder exposed to osmotic stress. The $H_2O_2$ concentration and the SOD and CAT expression and activity levels were lower in flounder fed with Diets 2 and 3 than with Diet 1, suggesting that quercetin directly scavenges reactive oxygen species to reduce oxidative stress. Furthermore, the plasma lysozyme activity and osmolality were higher with Diets 2 and 3 than with Diet 1, indicating that quercetin increases immune function and helps to maintain physiological homeostasis. Plasma cortisol was lower with Diets 2 and 3 than with Diet 1, suggesting the quercetin protects against stress. These results indicate that quercetin has hypocholesterolemic and antioxidant effects, increases immune function, and acts to maintain physiological homeostasis.

• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.53-57
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• 2004

The study was carried out to investigate the effects of morphology, reproductive cycle, incubation time and activation of oocytes on in vitro maturation of cat oocytes and development of IVM/IVF embryos. The results were summarized as follows: 1. When recovered from ovaries collected at different stages of the reproductive cycle (inactive, follicular and luteal stage), the developmental rates of oocytes to GV and MI stage were 72.5% and 27.5%, 57.5% and 7.5%, 62.5% and 17.5%, respectively. 2. The developmental rates of oocytes with cumulus cells to GV and MI stage in different conditions of incubation (5% $CO_2$ , 95% $O_2$ and 10% $CO_2$, 90% $O_2$) were 70.0% and 27.5%, 52.5% and 20.0%, 55.0% and 12.5%, respectively. 3. The developmental rates to GV and MI oocytes when cultured at different time of incubation (17∼20, 21∼24, 25∼28 and 29∼32 h) were 67.5% and 20.0%, 67.5% and 30.0%, 62.5% and 22.5%, 65.0% and 15.0%, respectively. 4. The fertilization and cleavage rates of freshly collected oocytes with and without cumulus cells were 72.5% and 25.0%, 37.5% and 7.5%, respectively. The rates were greater in oocytes with cumulus cells than those without cumulus cells. 5. The fertilization and cleavage rates of oocytes recovered from ovaries collected at different stages of the reproductive cycle (inactive, follicular and luteal stage) were 75.0% and 25.0%, 40.0% and 7.5%, 50.0% and 15.0%, respectively.

• Journal of Life Science
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• v.15 no.4 s.71
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• pp.626-632
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• 2005

The polyamines are essential components of all eukaryotic cells and absolutely necessary for cell growth. In the present study, the cytoprotective role of polyamine was characterized. When $Ac_2F$ rat liver cells were treated with 1M 2,2'-azobis (2-amidinopropane) dehydrochloride (AAPH), a water soluble free radical initiator, viability of the cells was noticeably decreased due to the increase of reactive oxygen species (ROS). The cytotoxic effect of AAPH as well as ROS generation were significantly inhibited by the treatment of polyamines. Among polyamines, especially spermine at $20{\mu}M$ concentration exerted over $45\%$ inhibition of AAPH-induced ROS generation. Western blotting was performed to determine whether superoxide dismutase(SOD) or catalase (CAT) expression was involved in oxidative stress. The AAPH treatment blocked both SOD and CAT protein expressions. Spermine could recover those protein expressions to the untreated control levels. According to the result of cycline E measurement, AAPH might block the entry of the cells into S phase of the cell cycle. The reduced expression of cyclin E protein could be fully recovered by the addition of spermine. The antioxidative effects of spermine was also further proved by the apopotitic morphological analysis using ethidium bromide and acridine orange.