• Journal of Microbiology and Biotechnology
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• v.26 no.9
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• pp.1570-1578
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• 2016

Commercial application of laccase is often hampered by insufficient enzyme stocks, with very low yields obtained from natural sources. This study aimed to improve laccase production by mutation of a Coriolopsis gallica strain and to determine the biological properties of the mutant. The high-yield laccase strain C. gallica TCK was treated with N-methyl-N-nitro-N-nitrosoguanidine and ultraviolet light. Among the mutants isolated, T906 was found to be a high-production strain of laccases. The mutant strain T906 was stabilized via dozens of passages, and the selected ones were further processed for optimization of metallic ion, inducers, and nutritional requirements, which resulted in the optimized liquid fermentation medium MF9. The incubation temperature and pH were optimized to be 30℃ and 4.5, respectively. The mutant strain T906 showed 3-times higher laccase activity than the original strain TCK under optimized conditions, and the maximum laccase production (303 U/ml) was accomplished after 13 days. The extracellular laccase isoenzyme 1 was purified and characterized from the two strains, respectively, and their cDNA sequence was determined. Of note, the laccase isoenzyme 1 transcription levels were overtly increased in T906 mycelia compared with values obtained for strain TCK. These findings provide a basis for C. gallica modification for the production of high laccase amounts.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.273-278
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• 2002

Two-step fed-batch fermentations were carried out to overproduce Bacillus licheniformis maltogenic amylase (BLMA) in recombinant Escherichia coli. The first step was to increase the cell mass by controlling the feeding of a glucose solution, while the second step was designed to improve the amylase expression efficiency by supplementing organic nitrogen sources. The linear gradient feeding method was successfully adopted to maintain the glucose concentration below 0.2 g/l during the fed-batch mode, as effectively minimizing acetic acid formation. When the dissolved oxygen (DO) level became limiting, an accumulation of acetic acid and drastic decrease in specific BLMA productivity were observed. Glucose and organic nitrogen sources consisting of yeast extract and casein hydrolysate were simultaneously supplied in the pH-stat mode to further increase the specific BLMA expression efficiency. An organic nitrogen source consisting of 200 g/1 yeast extract and 100 g/1 casein hydrolysate was found to be the best among the various combinations tested. The feeding of an organic nitrogen source in the second-step fed-batch period was highly beneficial in enhancing the BLMA production. The optimized two-step fed-batch culture resulted in 78 g/l maximum dry cell mass and 443 U/ml maximum BLMA activity, corresponding to 1.5-fold increase in the dry cell mass and 3.7-fold enhancement in BLMA production, compared with the simple fed-batch fermentation.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.301-305
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• 2002

The gene (ppaT) encoding Thermus caldophilus GK24 pyrophosphatase (Tca pyrophosphatase) was cloned and sequenced. The gene was found to contain an open reading frame encoding 175 amino acids with a calculated mass of 19,155 Da. The ppaT gene was expressed under the control of the tac promoter in Escherichia coli. The recombinant Tca pyrophosphatase was purified 21.4-fold with $56\%$ yield and specific activity of 25.7 U $mg^-1$, following a combination of heating (to denature the E. coli proteins) and one step of DEAE-Sephacel column chromatography. The native enzyme was found to have an approximate molecular mass of 110,000 Da and consisted of six subunits. The enzyme exhibited maximal activity at pH of 8.0-8.5 and was stable at $80-90^{\circ}C$. A divalent cation was absolutely required for the enzyme activity, with $Mg^2+$. being the most effective.

• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1753-1759
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• 2006

Paenibacillus sp. HY-8 isolated from the digestive tracts of the longicorn beetle, Moechotypa diphysis, produced an extracellular endoxylanase with a molecular weight of 20 kDa estimated by SDS-PAGE. The xylanase was purified to near electrophoretic homogeneity from the culture supernatant after ammonium sulfate precipitation, gel filtration, and ionexchange chromatography. The purified xylanase exhibited the highest activities at pH 6.0 and $50^{\circ}C$. The $K_m\;and\;V_{max}$ values were 7.2 mg/ml and 16.3 U/mg, respectively, for birchwood xylan as the substrate. Nucleotide sequence of the PCR-cloned gene was determined to have the open reading frame encoding a polypeptide of 212 amino acids. The N-terminal amino acid sequence and the nucleotide sequence analyses predicted that the precursor xylanase contained a signal peptide composed of 28 amino acids and a catalytically active 19.9-kDa peptide fragment. The deduced amino acid sequence shared extensive similarity with those of the glycoside hydrolase family 11 of xylanases from other bacteria. The predicted amino acid sequence contained two glutamate residues, previously identified as essential and conserved for active sites in other xylanases of the glycoside hydrolase family 11.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1863-1870
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• 2015

Fibrinolytic enzyme genes (aprE2, aprE176, and aprE179) were introduced into the Bacillus subtilis 168 chromosome without any antibiotic resistance gene. An integration vector, pDG1662, was used to deliver the genes into the amyE site of B. subtilis 168. Integrants, SJ3-5nc, SJ176nc, and SJ179nc, were obtained after two successive homologous recombinations. The integration of each fibrinolytic gene into the middle of the amyE site was confirmed by phenotypes (Amy-, Spec^S) and colony PCR results for these strains. The fibrinolytic activities of the integrants were higher than that of B. subtilis 168 by at least 3.2-fold when grown in LB broth. Cheonggukjang was prepared by inoculating each of B. subtilis 168, SJ3-5nc, SJ176nc, and SJ179nc, and the fibrinolytic activity of cheonggukjang was 4.6 ± 0.7, 10.8 ± 0.9, 7.0 ± 0.6, and 8.0 ± 0.2 (U/g of cheonggukjang), respectively at 72 h. These results showed that construction of B. subtilis strains with enhanced fibrinolytic activities is possible by integration of a strong fibrinolytic gene via a marker-free manner.

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.776-783
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• 2018

The agarase gene gaa16a was identified from a draft genome sequence of Gilvimarinus agarilyticus JEA5, an agar-utilizing marine bacterium. Recently, three agarase-producing bacteria, G. chinensis, G. polysaccharolyticus, and G. agarilyticus, in the genus Gilvimarinus were reported. However, there have been no reports of the molecular characteristics and biochemical properties of these agarases. In this study, we analyzed the molecular characteristics and biochemical properties of agarases in Gilvimarinus. Gaa16A comprised a 1,323-bp open reading frame encoding 441 amino acids. The predicted molecular mass and isoelectric point were 49 kDa and 4.9, respectively. The amino acid sequence of Gaa16A showed features typical of glycosyl hydrolase family 16 (GH16) ${\beta}$-agarases, including a GH16 domain, carbohydrate-binding region (RICIN domain), and signal peptide. Recombinant Gaa16A (excluding the signal peptide and carbohydrate-binding region, rGaa16A) was expressed as a fused protein with maltose-binding protein at its N-terminus in Escherichia coli. rGaa16A had maximum activity at $55^{\circ}C$ and pH 7.0 and 103 U/mg of specific activity in the presence of 2.5 mM $CaCl_2$. The enzyme hydrolyzed agarose to yield neoagarotetraose as the main product. This enzyme may be useful for industrial production of functional neoagaro-oligosaccharides.

• Korean Journal of Oriental Medicine
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• v.18 no.3
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• pp.119-126
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• 2012

1. Objectives Sasang constitutional medicine is unique in Korean traditional medicine. It diagnoses and treats patients based on his/her Sasang constitution (SC). Skin properties have been used as an effective diagnostic component in the classification of SC types in clinics. In this paper, we investigated the SC-based health relevance of skin elasticity and wrinkle properties. 2. Methods The skin elasticity and wrinkle of forearm and dorsal hand were measured in 299 elderly female subjects. To determine the subject's Sasang constitution, we adopted the classification results from a newly developed SC diagnostic tool. The health states of the subjects were scored by two Korean traditional medical doctors, by whom each subject was categorized either into the healthy state or the unhealthy state. 3. Results As a result, the elasticity hysteresis of forearm (E_HYS), the visco-elasticity (VE_MEAN), and the wrinkle frequency energy of backhand (W_HAND) showed significant differences between Taeum-in group and Soeum-in group. In case of the Soeum-in on unhealthy state, VE_MEAN was decreased significantly (p<.05). W_HAND and W_ARM_H of the healthy Taeum-in were less than those of the unhealthy Taem-in. 4. Conclusions In this study we showed that, for an elderly female population, skin elasticity and viscosity were significantly different not only between each SC type but also between healthy group and unhealthy group in each constitution. In particular, Soeum-in subjects were inferred to be superior in retaining skin softness when they were healthy, and Taeum-in subjects were easy to lose their firmness of skin surface when they became unhealthy.

• Journal of Trauma and Injury
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• v.33 no.2
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• pp.96-103
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• 2020

Purpose: In patients with trauma, rhabdomyolysis (RM) can lead to fatal complications resulting from muscle damage. Thus, RM must be immediately diagnosed and treated to prevent complications. Creatine kinase (CK) is the most sensitive marker for diagnosing RM. However, relying on CK tests may result in delayed treatment, as it takes approximately 1 hour to obtain CK blood test results. Hence, this study investigated whether the neutrophil-to-lymphocyte ratio (NLR) could predict RM at an earlier time point in patients with trauma, since NLR results can be obtained within 10 minutes. Methods: This retrospective study included 130 patients with severe trauma who were admitted to the emergency room of a tertiary institution between January 2017 and April 2020. RM was defined as a CK level ≥1,000 U/L at the time of arrival. Patients with severe trauma were categorized into non-RM and RM groups, and their characteristics and blood test results were analyzed. Statistical analysis was performed using SPSS version 26.0 for Windows. Results: Of the 130 patients with severe trauma, 50 presented with RM. In the multivariate analysis, the NLR (odds ratio [OR], 1.252; 95% confidence interval [CI], 1.130-1.386), pH level (OR, 0.006; 95% CI, 0.000-0.198), presence of acute kidney injury (OR, 3.009; 95% CI, 1.140-7.941), and extremity Abbreviated Injury Scale score (OR, 1.819; 95% CI, 1.111-2.980) significantly differed between the non-RM and RM groups. A receiver operating characteristic analysis revealed that a cut-off NLR value of 3.64 was the best for predicting RM. Conclusions: In patients with trauma, the NLR at the time of arrival at the hospital is a useful biochemical marker for predicting RM.

• Journal of Environmental Health Sciences
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• v.11 no.2
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• pp.17-27
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• 1985

The objectives of this study is to examine efficiency of swinery wastewater treatment by trickling filters' pilot plant. The results of this study are as follows: 1. The characteristics of sample. The BOD$_5$ was from 2,450 to 2,880mg/l, COD(KMnO$_4$ acid method) was from 910 to 1,064mg/l, and SS was from 920 to 990mg/l. The pH of influent was from 7.3 to 7.6, and the temperature of water was from 17.0$\circ$C to 22.5$\circ$C. 2. For experiment by recirculation, the BODs removal efficiency is 65.2% at recirculation ratio (r)=0, and 70.7% at r=1. The ramoval efficiency of this study is higher than NRC formula of U.S.A.. The recirculation is not significant effect on removal efficiency. 3. For experiment by hydraulic load, the BOD$_5$ removal rate decreased from 73.1% at $3.1m^3/m^2\cdot d$ to 65.3% at $9.2m^3/m^2\cdot d$. The design formula of this study which shows the removal rate of soluble BOD is $Le/Li =10^{-0.24} D/Q^{0.24}$ (Q: hydraulic load, D: depth of filter). 4. For experiment by organic load, the BOD$_5$ removal rate is increased from 70.2% at $0.77kg/m^3\cdot d$ to 75.4% at $4.28kg/m^3\cdot d$. We can obtain the straight line y=0.749 x (y: removed BOD, x :applied BOD) by the least squares method. 5. We can know that trickling filters is strong with the hydraulic load and the organic matter shock load. Here, we can judge that trickling filters is a good method for the treatment of swinery wastewater which containing high concentrated organic matter.

• Journal of Technologic Dentistry
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• v.23 no.2
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• pp.211-221
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• 2002

The purpose of this study was to analyze the commercial paste opaque products currently available in domestic of foreign as well as domestic, such as Duceram Plus(DU; Ducera Dental GmbH, Germany), VMK 95(VM; Vita Co., Germany), Noritake EX-3(EX; Noritake Co., Japan) and Ceramax(CE; Alphadent Co., Korea). They were characterized in thermal expansion coefficient, particle size distribution, viscosity and solvent using thermomechanical analyzer, particle size analyzer, rheometer and infrared spectrophotometer. Experimental results are as follows; Firstly, thermal expansion coefficients were determined $13.9{\times}10-6/^{\circ}C$ for DU, $14.3{\times}10-6/^{\circ}C$ for VM, $13.3{\times}10-6/^{\circ}C$ for EX, and $14.0{\times}10-6/^{\circ}C$ for CE. Secondly, percent of partice size below $1{\mu}m$ were 12% for DU, VM and CE, and 13% for EX, percent between $1{\mu}m$ and $5{\mu}m$ were 42% for DU, 42% for VM, 38% for EX, and 61 % for CE, percent between $5{\mu}m$ and $10{\mu}m$ were 21 % for DU, 24% for VM, 20% for EX, and 18% for CE, and over $10{\mu}m$ were 25% for DU, 22% for VM, 29% for EX, 9% for CE. Thirdly, the basic composition of the solvent in all of the commercial paste opaques were determined as ethylene glycol from FT-IR investigation. Lastly, measured viscosities were 1798 cp for DU, 536 cp for VM, 1110 cp for EX, and 721 cp for CE.