• Proceedings of the Korean Society for Applied Microbiology Conference
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• 한국미생물생명공학회 1978년도 춘계학술대회
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• pp.96.3-96
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• 1978

Biotin 요구성 glutamic acid 생산능을 가진 Br-evibacterium sp.의 isoleucine 영양요구성 변이주가 다량의 L-leucine을 생성함을 발견하였다. 본균주에 관한 탄소원, 질소원, 무기염, vitamin, amino acid 등의 영양물질과 온도, 공기주입량, pH등에 관한 실험으로 다음과 같은 결과를 얻었다. 본균주는 탄소원, 질소원으로서 10% glucose와 4% $(NH_4)_2SO_4를$ 공합하여 $30^{\circ}C,$ 88시간 배양시킨 결과, 액중에 21 mg/ml의 L-leucine이 축적되었다. Fe$^{2+}$와 $Mn^{2+}$의 동시첨가는 L-leucine생성에 효과적이었으며 thiamine 역시 L-leucine 생성에 효과적이었다. 30ι jar fermentor 에서 폐당밀을 주원료로 한 실험결과, 폐당밀, $(NH_4)/SO_4,$ $KH_2PO_4,$ $MgSO_4ㆍ7H_2O로$ 구성된 액체배지에서 $30^{\circ}C,$ 36시간 배양시킨 것이, 27mg/ml의 L-leucine이 축적되었으며 이때의 대당수율은 16%이었다.

• Proceedings of the Korean Vacuum Society Conference
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• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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• pp.263.2-263.2
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• 2013

We report a systematic investigation of a set of macrophotoinitiators for use in polymerization-based signal amplification. To test the dependence of photopolymerization responses on the number of photoinitiators localized per molecular recognition event, we gradually increased the number of photoinitiator molecules coupled to a scaffold macromolecule. Macrophotoinitiators constructed with an average of 7 to 168 photoinitiators per polymer with the goals of quantifying the relationship between the number of initiators per binding event and the degree of amplified colorimetric readout. To evaluate the capacity of the macrophotoinitiators to detect molecular recognition, neutravidin was coupled to these molecules to recognize biotin-labeled DNA immobilized on biochip test surfaces. Fluorescein macroinitiators are found to be useful in detecting molecular recognition above a threshold of initiators per polymer. Above this threshold, increasing the number of initiators per macroinitiator resulted in increased signal strength.

• Animal Bioscience
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• 제34권10호
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• pp.1579-1589
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• 2021

Objective: This study was conducted to generate single stranded DNA oligonucleotides with selective affinity to bovine spermatozoa, assess its binding potential and explore its potential utility in trapping spermatozoa from suspensions. Methods: A combinatorial library of 94 mer long oligonucleotide was used for systematic evolution of ligands by exponential enrichment (SELEX) with bovine spermatozoa. The amplicons from sixth and seventh rounds of SELEX were sequenced, and the reads were clustered employing cluster database at high identity with tolerance (CD-HIT) and FASTAptamer. The enriched nucleotides were predicted for secondary structures by Mfold, motifs by Multiple Em for Motif Elicitation and 5' labelled with biotin/6-FAM to determine the binding potential and binding pattern. Results: We generated 14.1 and 17.7 million reads from sixth and seventh rounds of SELEX respectively to bovine spermatozoa. The CD-HIT clustered 78,098 and 21,196 reads in the top ten clusters and FASTAptamer identified 2,195 and 4,405 unique sequences in the top three clusters from the sixth and seventh rounds, respectively. The identified oligonucleotides formed secondary structures with delta G values between -1.17 to -26.18 kcal/mol indicating varied stability. Confocal imaging with the oligonucleotides from the seventh round revealed different patterns of binding to bovine spermatozoa (fluorescence of the whole head, spot of fluorescence in head and mid- piece and tail). Use of a 5'-biotin tagged oligonucleotide from the sixth round at 100 pmol with 4×10⁶ spermatozoa could trap almost 80% from the suspension. Conclusion: The binding patterns and ability of the identified oligonucleotides confirms successful optimization of the SELEX process and generation of aptamers to bovine spermatozoa. These oligonucleotides provide a quick approach for selective capture of spermatozoa from complex samples. Future SELEX rounds with X- or Y- enriched sperm suspension will be used to generate oligonucleotides that bind to spermatozoa of a specific sex type.

• KSBB Journal
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• 제8권2호
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• pp.156-163
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• 1993

Strains degrading ethylene glycol(EG) and terephthalic acid(TPA) were isolated from water systems, and identified as Pseudomonas sp. They were named as Pseudomonas sp. EAW for EG and as Pseudomonas sp. TS2 for TPA. The optimal culture conditions of temperature, pH and nitrogen source were found to be $35^{\circ}C$, 7.5 and ammonium sulfate, respectively. The growth of strains and removal efficiency was slightly promoted by trace elements such as niacin and biotin in case of EG, and by trace elements such as $Na_2MoO_4{\cdot}2H_2O$ and thiamin i case of TPA. With increasing inoculation sloe for batch culture, the removal efficiency of EG by the strain EAW was conspicuously increased, while the removal efficiency of TPA by the strain TS2 was not changed as much as that of EG. The growth rate of the strain EAW was much more decreased than that of the strain TS2 in the enrichment medium, as the frequency of repeated-batch culture in the rich-medium increased. in case of real wastewater, growth rate and removal efficiencies of EG and TPA were lower than those in the enrichment medium. $COD_{Mn}\;and\;COD_{Cr}$ removal efficiencies after 48 hrs batch culture in real wastewater were 89% and 93%, respectively. The specific growth rate was inhibited when the initial concentration of EG or TPA was more than 25g/L.

• Tuberculosis and Respiratory Diseases
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• 제62권1호
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• pp.43-50
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• 2007

Background: Mutated or deregulated expression of C-erbB-2 causes this gene to function as a potent oncogene. Vascular endothelial growth factor (VEGF) is a crucial angiogenic molecule in lung cancer. Both C-erbB-2 and VEGF can promote growth, proliferation and metastasis in non-small cell lung cancer (NSCLC). The purpose of this study was to investigate evaluate the relationship between the expressions of the C-erbB-2 and VEGF genes using immunohistochemistry. Materials and Methods: Ninety-five patients with NSCLC were involved (60 squamous cell carcinoma and 35 adenocarcinoma). The formalin-fixed paraffin embedded specimens were immunohistochemically stained for C-erbB-2 and VEGF using the avidin-biotin complex method. Results: Positive C-erbB-2 expression was observed more often in adenocarcinomas than squamous cell carcinomas (p<0.05). Although the immunohistochemical expressions of C-erbB-2 and VEGF in non-small-cell lung cancer showed increased tendencies at an advanced stage, the correlation between early and advanced cancers was insignificant. In adenocarcinomas, the expressions of VEGF and C-erbB-2 were significantly (p<0.05). Conclusion: The overexpression fo C-erbB-2 was significantly higher in adenocarcinomas than squamous cell carcinomas, and correlated with the expression of VEGF in adenocarcinomas of the lungs.

• Journal of Mushroom
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• 제2권4호
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• pp.207-213
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• 2004

The results of examining characteristics of mycelial growth and culture condition for determining the condition of artificially culturing Fomitopsis pinicola are as follows. 1) Mycelial growth and density of F. pinicola. were the highest in the medium of PIDA(Pine Dextrose Agar;66.3mm/10d) followed by the order of GDA, PDA, CDA, PODA, ODA, YM, MCM, MEA(pH 4.7), CHA, and MEA(pH 4.7). 2) Optimal temperature for the mycelial growth and density of F. pinicola were shown to be $30^{\circ}C$, but the mycelia were dead at $40^{\circ}C$. the mycelial growth and density of KNAC9005 strains was the highest at $30^{\circ}C$(66.3mm/10d) followed by the order of 25, 20, 15, 35, 10, and $5^{\circ}C$. 3) Optimal pH for the mycelial growth and density of $40^{\circ}C$ was revealed to be 6.0(88.4mm/10d). above or below pH 6.0, the mycelial growth and density were shown to be retarded. 4) Optimal carbon, nitrogen and organic acid sources for the spawn growth of $40^{\circ}C$ were maltose(331mg/25ml/15d), peptone(347mg/25ml/15d), and glutamic acid(357mg/25ml/15d), respectively. Optimal level of biotin was 370mg/15d and optimal C/N ratio was 40.

• Journal of Microbiology and Biotechnology
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• 제23권2호
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• pp.211-217
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• 2013

Corynebacterium glutamicum is one of the well-studied industrial strain that is used for the production of nucleotides and amino acids. Recently, it has also been studied as a possible producer of organic acids such as succinic acid, based on its ability to produce organic acids under an oxygen deprivation condition. In this study, we conducted the optimization of medium components for improved succinate production from C. glutamicum under an oxygen deprivation condition by Plackett-Burman design and applied a response surface methodology. A Plackett-Burman design for ten factors such as glucose, ammonium sulfate, magnesium sulfate, potassium phosphate ($K_2HPO_4$ and $KH_2PO_4$), iron sulfate, manganese sulfate, biotin, thiamine, and sodium bicarbonate was applied to evaluate the effects on succinate production. Glucose, ammonium sulfate, magnesium sulfate, and dipotassium phosphate were found to have significant influence on succinate production, and the optimal concentrations of these four factors were sequentially investigated by the response surface methodology using a Box-Behnken design. The optimal medium components obtained for achieving maximum concentration of succinic acid were as follows: glucose 10 g/l, magnesium sulfate 0.5 g/l, dipotassium phosphate ($K_2HPO_4$) 0.75 g/l, potassium dihydrogen phosphate ($KH_2PO_4$) 0.5 g/l, iron sulfate 6 mg/l, manganese sulfate 4.2 mg/l, biotin 0.2 mg/l, thiamine 0.2 mg/l, and sodium bicarbonate 100 mM. The parameters that differed from a normal BT medium were glucose changed from 40 g/l to 10 g/l, dipotassium phosphate ($K_2HPO_4$) 0.5 g/l changed to 0.75 g/l, and ammonium sulfate ($(NH_4)_2SO_4$) 7 g/l changed to 0 g/l. Under these conditions, the final succinic acid concentration was 16.3 mM, which is about 1.46 fold higher than the original medium (11.1 mM) at 24 h. This work showed the improvement of succinate production by a simple change of media components deduced from sequential optimization.

• Korean Journal of Plant Resources
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• 제15권2호
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• pp.135-143
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• 2002

The results of examining characteristics of mycelial growth and culture condition for determining the condition of artificially culturing Inonotus obliqua (chaga) are as follows. 1) Mycelial growth and density oft, oblique were the highest in the medium of BDA (Birch Dextrose Agar; 66.3mm/10d) followed by the order of GDA, PDA, CDA, PODA, ODA, YM, MCM, MEA (pH 7.0), CHA, and MEA (pH 4.7). 2) Optimal temperature for the mycelial growth and density of 1. obliqua were shown to be 30$^{\circ}C$, but the mycelia were dead at 40$^{\circ}C$. the mycelial growth and density of KNAC3005 strains was the highest at 30$^{\circ}C$(66.3mm/10d) followed by the order of 25, 20, 15, 35, 10, and 5$^{\circ}C$. 3) Optimal pH for the mycelial growth and density were revealed to be 6.0 (88.4mm/10d). Above or below pH 6.0, the mycelial growth and density were shown to be retarded. 4) Optimal carbon, nitrogen and organic acid sources for the spawn growth of 40 were maltose (331mg/25$m\ell$/15d), peptone (347mg/25$m\ell$/15d), and glutamic acid (357mg/25$m\ell$/15d), respectively. Optimal level of biotin was 370mg/15d and optimal C/N ratio was 40.

• Journal of the Korean Society of Food Science and Nutrition
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• 제16권4호
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• pp.251-261
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• 1987

This study was attempted to increase the production of L-Threonine by Brevibacterium Flavum ATCC 14067, To select the strain which produce the highest threonine, mutants ere induced by N-methyl-N'-nitro-N-nitrosoguanidine treatment. The composition of media and cultural condition for its overproduction of threonine were also studied. In a threonine producer, strain B-13(Met-) was the strain producing the highest amount of threonige among methionine, lysine and isoleucine auxotrophs. The following results were obtained. 1. The wild strain and B-13(Met-) produced threonine 1.4mg/ml and 4.86mg/ml , respectively. 2. The optimum composition of medium for producing threonine by Brevibacterium Flavum B-13 was glucose 10%, ammonium sulfate 4%, potassium phosphate monobasic 0.2%, magnesium sulfate 0.05%, biotin $200{\mu}l$, thiamine $300{\mu}l$. Addition of nicotinic acid also led to increase L-threonine production. 3. In addition of organic nutrients to the fermentation medium, peptone n'ere effective and addition of methionine $100{\mu}g/ml$ produced the highest amount of L-Threonine. Aspartic acid and homoserine were also effective when these amino acid were added to the fermentstion medium. 4. Cultural conditon on threonine production by B-16 were investigated. The optimum pH was 7.0-8.0. The highest amount of threnine was produced after 4 days of cultural period.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.656-660
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• 2013

Pseudomonas fluorescens strain CL0145A was discovered at the New York State Museum Field Research Laboratory as an effective agent against the environmentally destructive zebra mussel, which has contaminated US waters. Dried cells of the microbe are being commercialized as an environmentally friendly solution to the problem. We found that antibiotic activity against the Gram-positive bacterium Bacillus subtilis is produced and excreted by this strain. We have carried out studies to optimize production of the antibiotic. Studies were begun in a complex corn meal medium. Activity was found in both cells and culture supernates and was maximal after one day of fermentation. Static fermentation conditions were found to be superior to shaken culture. Production of extracellular antibiotic in complex medium was found to be dependent on the content of sucrose and enzyme-hydrolyzed casein. Indeed, production was greater in sucrose plus enzyme-hydrolyzed casein than in the complex medium. Of a large number of carbon sources studied as improvements over sucrose, the best was glycerol. An examination of nitrogen sources showed that production was improved by replacement of enzyme-hydrolyzed casein with soy hydrolysates. Production in the simple glycerol-Hy-Soy medium was not improved by addition of an inorganic salt mixture or by complex nitrogen sources, with the exception of malt extract. In an attempt to keep the medium more defined, we studied the effect of amino acids and vitamins as replacements for malt extract. Of 21 amino acids and 7 vitamins, we found tryptophan, glutamine, biotin, and riboflavin to be stimulatory. The final medium contained glycerol, Hy-Soy, tryptophan, glutamine, biotin, and riboflavin.