• Journal of Life Science
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• v.18 no.11
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• pp.1592-1599
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• 2008

The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.

• Progress in Medical Physics
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• v.25 no.4
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• pp.199-204
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• 2014

The purpose of this study is to evaluate the dosimetric outcome of the field-in-field (FIF) plans compared with tangential wedged beams (TWB) plans for whole breast irradiation of breast cancer patients. Twenty patients with right-sided breast cancer and 10 patients with left-sided breast cancer were retrospectively enrolled in this study. We generated a FIF plan and a TWB plan for each patient to compare dosimetric outcomes. The dose the homogeneity index (HI), the conformity index (CI) and the uniformity index (UI) were defined and used for comparison of the dosimetric outcome of the planning target volume (PTV). To compare the dosimetric outcome of the organs at risk, the mean dose ($D_{mean}$) and the percentage of volumes receiving more than 10, 20 and 30 Gy of the ipsilateral lung and heart were used. The FIF plans had significantly lower HI (p=0.002), higher UI (p=0.000) and CI (p=0.000) than those of the TWB plans, which means that the FIF plans were better than the TWB plans in the dosimetric comparisons of the PTV. The $V10_{lung}$ ($17.1{\pm}7.1$ vs. $18.6{\pm}6.6%$, p=0.020) and $V30_{lung}$ ($10.3{\pm}5.1%$ vs. $10.7{\pm}5.2%$, p=0.000) were lower with the FIF plans compared with those of the TWB plans, with statistical significance. For the left-sided breast cancer patients, $D_{mean}$ of the heart ($2.6{\pm}1.3$ vs. $3.2{\pm}1.4$ Gy, p=0.000), $V20_{heart}$ ($3.4{\pm}2.6$ vs. $3.6{\pm}2.8%$, p=0.005) and $V30_{heart}$ ($2.6{\pm}2.3%$ vs. $2.9{\pm}2.4%$, p=0.004) were significantly lower for the FIF plans in comparison with those of the TWB plans. The FIF plans increased the dose homogeneity, conformity and uniformity of the target volume for the whole-breast irradiation compared with the TWB plans. Moreover, FIF plans reduced the doses to the ipsilateral lung and heart.

• Journal of Microbiology and Biotechnology
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• v.28 no.4
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• pp.588-596
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• 2018

An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.

• Korean Journal of Ecology and Environment
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• v.38 no.2 s.112
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• pp.137-145
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• 2005

Traditional screening techniques have missed up to 99% of microbial resources existing in the nature. Strategies of direct cloning of environmental DNAs comprising tine genetic blueprints of entire microbial metagenomes provide vastly more genetic information than is contained in the culturable. Therefore, one way to screening the useful gene in a variety of environments is the construction of metagenomic DNA library. In this study, the water samples were collected from Daecheong Reservoir in the mid Korea, and analyzed by T-RFLP to examine the diversity of the microbial communities. The crude DNAs were extracted by SDS-based freezing-thawing method and then further purified using an $UltraClean^{TM}kit$ (MoBio, USA). The metagenomic libraries were constructed with the DNAs partially digested with EcoRI, BamHI, and SacII in Escherichia coli DH10B using the pBACe3.6 vector. About 14.0 Mb of metagenomic libraries were obtained with average inserts 13 ${\sim}$ 15 kb in size. The genes responsible for degradation of 1, 2-dichloroethane (1, 2-DCE) via hydrolytic dehalogenation were identified from the metagenomic libraries by colony hybridization. The 1, 2-dichloroethane dehalogenase gene (dhlA) was cloned and its nucleotide sequence was analyzed. The activity of the 1, 2-DCE dehalogenase was highly expressed to the substrate. These results indicated that the dhlA gene identified from the metagenomes derived from Deacheong Reservoir might be useful to develop a potent strain for degradation of 1, 2-DCE.

• Journal of Life Science
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• v.18 no.1
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• pp.84-90
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• 2008

Bacterial colonies which were able to oxidize the manganese were isolated from six soil samples in Byungchon area. Among them, one bacterial strain was selected for this study based on its high manganese oxidation activity. This selected bacterial strain was identified as Pseudomonas sp. MN5 through physiological-biochemical test and analysis of its 16s rRNA sequence. This selected bacterial strain was able to utilize fructose and maltose, but they doesn't utilizing various carbohydrates as a sole carbon source. Pseudomonas sp. MN5 showed a very sensitive to antibiotics such as kanamycin, chloramphenicol, streptomycin and tetracycline, but a high resistance up to mg/ml unit to heavy metals such as lithium, manganese and barium. Optimal manganese oxidation condition of Pseudomonas sp. MN5 was pH 7.5 and manganese oxidation activity was inhibited by proteinase K and boiling treatment. The manganese oxidizing protein produced by Pseudomonas sp. MN5 was purified by ammonium sulfate precipitation, HiTrap Q FF anion exchange chromatography and G3000sw $_{XL}$ gel filtration chromatography. By sodium dodecyl sulfate polyacrylamide gel electrophoresis, three manganese oxidizing protein with estimated molecular weights of 15 kDa, 46.7 kDa and 63.5 kDa were detected. Also, it was estimated that manganese oxidizing protein produced by Pseudomonas sp. MN5 were a kind of porin proteins through internal sequence and N-terminal sequence analysis.

• Journal of Life Science
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• v.21 no.2
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• pp.221-226
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• 2011

The gene encoding 4-$\alpha$-glucanotransferase (mgtA) from Thermotoga neapolitana was cloned and expressed in Escherichia coli in order to investigate whether this enzyme was capable of producing cycloamylose for industrial applications. MgtA was purified to homogeneity by HiTrap Q HP and Sephacryl S-200 HR column chromatographies. The size of the enzyme as determined by SDS-PAGE was about 52 kDa, which was in good agreement with its deduced molecular mass of 51.9 kDa. The optimal temperature and pH for the activity of the 4-$\alpha$-glucanotransferase was found to be $85^{\circ}C$ and 6.5, respectively. The enzyme hydrolyzed the 1,4-$\alpha$-glucosidic bonds in oligomeric 1,4-$\alpha$-glucans and transferred oligosaccharides (maltotriose being the shortest one) to acceptor maltodextrins. However, the enzymes had no activity against pullulan, glycogen, and other di- or trioligosaccharides with rare types of $\alpha$-bond. MgtA is distinguished from 4-$\alpha$-glucanotransferase from Thermotoga maritima in that it can convert maltotriose into maltooligosaccharides. The treatment of glucoamylase after the reaction of MgtA with maltotriose, maltotetraose, maltopentaose, or maltohexaose as sole substrate revealed that MgtA yielded linear maltooligosaccharides instead of cycloamylose.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.463-470
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• 2020

The JS18 strain was isolated from an old tree forest and produced extracellular enzymes that decolorize synthetic melanin. Phylogenetic analysis, based on the internal transcribed spacer (ITS) sequence, indicate that JS18 belongs to the Trametes velutina species. JS18 demonstrated laccase activity but no manganese peroxidase or lignin peroxidase activity. Batch culture indicated that the melanin decolorization activity of JS18 strain originated from the laccase. Syringic acid and CuSO4 induced maximum laccase production, yielding 98 U/ml laccase activity after cultivation for 7 days at 25℃. T. velutina secretes an extracellular laccase in GYP medium, and this enzyme was purified using (NH4)2SO4 precipitation, Hi-trap Q Sepharose columns and gel filtration. The molecular weight of the purified enzyme was estimated to be 67 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis. This enzyme produced 80% of its melanin decolorization activity within the first 24 h of evaluation in the presence of 1-hydroxybenzotriazole (HBT), while only about 4% of the melanin was decolorized in the absence of the mediator. The greatest decolorization was observed at 1.5 mM/l HBT, which decolorized 81% of the melanin within the first 24 h. The optimum pH and temperature for this decolorization were found to be 5.0 and 37℃, respectively. Our results suggest the possibility of applying HBT induced T. velutina JS18 laccase-catalyzed melanin decolorization.

• Phonetics and Speech Sciences
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• v.16 no.1
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• pp.67-76
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• 2024

Recent advances in text-to-speech (TTS) technology have significantly improved the quality of synthesized speech, reaching a level where it can closely imitate natural human speech. Especially, TTS models offering various voice characteristics and personalized speech, are widely utilized in fields such as artificial intelligence (AI) tutors, advertising, and video dubbing. Accordingly, in this paper, we propose a one-shot multi-speaker TTS system that can ensure acoustic diversity and synthesize personalized voice by generating speech using unseen target speakers' utterances. The proposed model integrates a speaker encoder into a TTS model consisting of the FastSpeech2 acoustic model and the HiFi-GAN vocoder. The speaker encoder, based on the pre-trained RawNet3, extracts speaker-specific voice features. Furthermore, the proposed approach not only includes an English one-shot multi-speaker TTS but also introduces a Korean one-shot multi-speaker TTS. We evaluate naturalness and speaker similarity of the generated speech using objective and subjective metrics. In the subjective evaluation, the proposed Korean one-shot multi-speaker TTS obtained naturalness mean opinion score (NMOS) of 3.36 and similarity MOS (SMOS) of 3.16. The objective evaluation of the proposed English and Korean one-shot multi-speaker TTS showed a prediction MOS (P-MOS) of 2.54 and 3.74, respectively. These results indicate that the performance of our proposed model is improved over the baseline models in terms of both naturalness and speaker similarity.

• Tuberculosis and Respiratory Diseases
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• v.44 no.1
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• pp.136-145
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• 1997

Background : Tracheal Gas Insufflation (TGI) is one of the newer ancillary measures in mechanical ventilation employed to enhance carbon dioxide elimination. TGI exerts its effect through reduction of deadspace ventilation, but the factors determining its effect are not well studied yet. Method : The subjects were seven mechanically-ventilated patients ($58.8{\pm}10.6$ yrs) who showed increased physiologic deadspace greater than 60%. After 30 nun of stabilization with 100% oxygen on pressure control ventilation, continuous flow TGI was administered via the insufflation lumen of Hi-Lo Jet Tracheal Tube (Mallincrodt, USA) for 15 min at 3 L/min and 5 L/min each. Results : $PaCO_2$ was decreased ($51.4{\pm}17.6$ at baseline, $49.1{\pm}18.9$ at TGJ 3 L/min $45.0{\pm}14.9$ mm Hg at TGI 5 L/min, p=0.050), and pH was increased ($7.37{\pm}0.12$, $7.38{\pm}0.13$, $7.39{\pm}0.12$, respectively, p=0.037) while mixed expired $CO_2$ ($P_ECO_2$) was not changed significantly from baseline (p=0.336) by TGI. Physiologic deadspace(Vdphy) was decreased ($73.0{\pm}7.9$% at baseline, $69.8{\pm}10.0$% at TGI 3 L/min, and $67.1{\pm}10.1$% at TGI 5 L/min, p=0.015). $AaDO_2$(p=0.147), Vt(p=0.2140), Pmean(p=0.7788) and mean arterial pressure(p=0.4169) were not changed. The correlation between % maximal decrease of Vdphy were r=0.790 with the ratio of baseline Vdana/Vdphy(p=0.035) and r=-0.754 with baseline Vdalv(p=0.050). Conclusion: TGI was effective in reducing $PaCO_2$ and deadspace, and the deadspace-reducing effect was best correlated with baseline anatomic/physiologic deadspace ratio.

• Journal of Acupuncture Research
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• v.22 no.1
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• pp.91-97
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• 2005

Objective : This study was designed to evaluate the effect of moxibustion with various scales on symptoms of idiopathic Parkinson's disease. Methods : Subjects were voluntarily recruited by newspapers and internet. All the subjects are confirmed as idiopathic parkinson's disease by a neurologist. The moxibustion therapy was performed 5 times a week by patient's family at home and once a week by oriental medical doctor at hospital. Moxibustion points were GV2O, CV12, ST36, BL18, BL2O. Intensity was up to pain threshold according to patients not to get burned. The patient's symptoms were assessed before, after 4 weeks and after 8 weeks treatment by unified Parkinson's disease rating scale(UPDRS), modified Hoehn-Yahr(H-Y) stage, Schwab & England activity of daily living and freezing of gait questionnaire(FOGQ). Results : Total UPDRS scores were significantly improved after 4 weeks(p<0.01) and after 8 weeks(p<0.01) compared to the pre-treatment. There were significant changes in H-Y stage after 4 weeks(p<0.05), but there were no significant changes in H-Y stage after 8 weeks. The scores of ADL were not significantly improved after 4 weeks(p>0.05) and after 8 weeks(p>0.05). There were significant changes in FOGQ scale after 4 weeks(p=0.05) and but there were no significant changes in FOGQ scale after 8 weeks(p=0.13). Conclusion : This study suggests that moxibustion treatments can be applicable to improve symptoms in the patients with idiopathic Parkinson's disease.