• Title/Summary/Keyword: strain selection

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Selection and Isolation of a Mutant Yeast Strain Tolerant to Multiple Targeted Heavy Metals

  • Lee, Sangman
    • Korean Journal of Environmental Agriculture
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    • v.33 no.2
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    • pp.129-133
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    • 2014
  • BACKGROUND: This study was performed for selecting yeast mutants with a high tolerance for targeted metals, and determining whether yeasts strains tolerant to multiple heavy metals could be induced by sequential adaptations. METHODS AND RESULTS: A mutant yeast strain tolerant to the heavy metals cadmium (Cd), copper (Cu), nickel (Ni), and zinc (Zn) was selected by sequential elevated exposures to each metal with intermittent mutant isolation steps. A Cd-tolerant mutant was isolated by growing yeast cells in media containing $CdCl_2$ concentrations that were gradually increased to 1 mM. Then the Cd-tolerant mutant was gradually exposed to increasing levels of $CuCl_2$ in growth media until a concentration of 7 mM was reached, thus generating a strain tolerant to both Cd and Cu. In the subsequent steps, this mutant was exposed to $NiCl_2$ (up to 8 mM), and a resultant isolate was further exposed to $ZnCl_2$ (up to 60 mM), allowing the derivation of a yeast mutant that was simultaneously tolerant to Cd, Cu, Ni, and Zn. CONCLUSION: This method of inducing tolerance to multiple targeted heavy metals in yeast will be useful in the bioremediation of heavy metals.

Comparative Study on the Nonlinear Material Model of HyperElastic Material Due to Variations in the Stretch Ratio (신장률 변화에 따른 초탄성 재료의 비선형 재료모델 비교 연구)

  • Lee, Kangsu;Ki, Minsuk;Park, Byoungjae
    • Journal of Ocean Engineering and Technology
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    • v.32 no.4
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    • pp.253-260
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    • 2018
  • Recently, the application of non-steel materials in ships and offshore plants is increasing because of the development of various nonlinear materials and the improvement of performance. Especially, hyper-elastic materials, which have a nonlinear stress-strain relationship, are used mainly in marine plant structures or ships where impact relaxation, vibration suppression, and elasticity are required, while elasticity must be maintained, even under high strain conditions. In order to simulate and evaluate the behavior of the hyperelastic material, it is very important to select an appropriate material model according to the strain of the material. This study focused on the selection of material models for hyperelastic materials, such as rubber used in the marine and offshore fields. Tension and compression tests and finite element simulations were conducted to compare the accuracy of the nonlinear material models due to variations in the stretch ratio of hyper-elastic material. Material coefficients of nonlinear material models are determined based on the curve fitting of experimental data. The results of this study can be used to improve the reliability of nonlinear material models according to stretch ratio variation.

Strain Improvement for Enhanced Production of Streptokinase and Streptodornase in Streptococcus sp.

  • HYUN, HYUNG-HWAN;YOON-BUM LEE;KYUNG-HWA SONG;JI-YOUNG JEON;HYUNE-HWAN LEE
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.101-106
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    • 1997
  • Strain improvement for the enhanced production of streptokinase and streptodornase in Streptococcus sp. ATCC 12449 was performed. Strain UB111, a hyperproductive mutant which was isolated by use of nitrosoguanidine and selection of colonies with large clear zones on DNase test agar plates supplemented with $1{\%}$ glucose and $0.5{\%}$ ammonium chloride, produced about 3 fold more streptokinase and streptodornase than the wild type when tested in shake flask fermentations. The enhanced production of both streptokinase and streptodornase was achieved by cultivating the mutant in a pH-controlled fermentor containing fermentation medium enriched with yeast extract ($2.1{\%}$). Under these conditions, the mutant produced 7300 units/ml of streptokinase and 800 units/ml of streptodornase.

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Fingerprinting of Listeria monocytogenes by Amplified Fragment Length Polymorphism Analysis

  • Jin, Hyun-Seok;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.8 no.1
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    • pp.29-37
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    • 2002
  • Listeria monocytogenes poses an increasing health risk, which in part is due to increasing health risk, consumption of ready-to-eat food products and the introduction of increasing numbers of food products from regions with different dietary habits. L. monocytogenes can be present in meat, shellfish, vegetables, unpasteurised milk and soft cheese and poses a risk if food containing these products is stored at refrigeration temperature and is not properly heated before consumption, as L. monocytogenes is psychrophilic. Amplified-fragment length polymorphism (AFLP) analysis is the method of genotypic techinique in which adaptor oligonucleotides are ligated to restriction enzyme fragments and then used as target sites for primers in a PCR amplification. The amplified fragments are electrophoretically separated to give strain-specific band profiles. Single-enzyme approach that did not require costly equipment or reagents for the fingerprinting of strains of Listeria monocytogenes was developed. Single-enzyme amplified fragment length polymorphism (SE-AFLP) analysis was used to perform species and strain identification of Salmonella, Shigella, Yersinia and E. coli. By careful selection of AFLP primers, it was possible to obtain reproducible and sensitive identification to strain level. The AFLP patterns of L. monocytogenes are divided by the kinds of specimens in which were isolated. SE-AFLP fragments can be analyzed using standard gel electrophoresis, and can be easily scored by visual inspection, due to the low complexity of the fingerprint obtained by this method. These features make SE-AFLP suitable for use in either field or laboratory applications.

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An Analysis of Poisson's Ratio Behaviors by Uniaxial Compressive Loading-reloading Test - On the Sedimentary Rocks of Kyungsang Basin - (일축압축 하에서 반복재하에 따른 포아송비의 거동분석 - 경상분지 퇴적암을 대상으로 -)

  • Lee, Jong-Suok;Moon, Jong-Kyu;Choi, Woong-Eui
    • Tunnel and Underground Space
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    • v.23 no.1
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    • pp.66-77
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    • 2013
  • This paper deals with Poisson's ratio and volumetric strain behavior on loading-reloading terms under uniaxial condition targeting 404 individual rocks, which include sedimentary rocks as sandstone, shale, mudstone, conglomerate and tuff on Kyungsang basin. Poisson' ratio demonstrates increase, convergence and decrease behavior according to the increase in load, which results in preponderance of increase behavior. Volumetric strain demonstrates normal, positive and negative behavior according to the increase in load, which results in preponderance of normal behavior. On practice, Poisson's ratio can be indicative of high or low values with low values of design load. Consequently, a careful selection of results in in-situ sample experiment should be made and varying design conditions should be considered.

Construction of a Plant Expression Vector for the Coat Protein Gene of Cucumber Mosaic Virus-As Strain for Plant Transformation (오이 모자이크 바이러스 As계통 외피단백질 유전자의 식물체 형질질환을 위한 발현벡타의 구축)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.11 no.1
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    • pp.66-72
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    • 1995
  • The coat protein (CP) gene of cucumber mosaic virus-As (CMV-As) strain was engineered for expression in the plant by using the cauliflower mosaic virus 35S transcript regulatory sequences. The CP gene was cloned into an Agrobacterium-derived binary vector. A chimeric gene was constructed by the cDNA of CMV-As CP and plant expression vector pBI121. The clone, pCMAS66, was first introduced into the phagemid vector pSPORT1 for situating sense orientation for translation and making restriction sites in order to re-introduce plant expression vector, pHI121. The resulting subclone pCASCP02 and plant expression vector pBI121 were treated with BamHI-SacI for excising the target gene and removing GUS gene, respectively. After Agrobacterium transformation by freeze-thaw technique, the clone, pCMASCP121-123 which contains sense orientation of the target gene, was selected and confirmed by restriction endonuclease analysis. The CMV-As CP gene was introduced into A. tumefaciens. The results on tobacco plant transformation with the vector system revealed that the system could be successfully introduced and showed high frequency of selection to putative transformations.

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Development of FK506-hyperproducing strain and optimization of culture conditions in solid-state fermentation for the hyper-production of FK506

  • Mo, SangJoon;Yang, Hyeong Seok
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.289-298
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    • 2016
  • FK506 hyper-yielding mutant, called the TCM8594 strain, was made from Streptomyces tsukubaensis NRRL 18488 by mutagenesis using N-methyl-N'-nitro-N-nitrosoguanidine, ultraviolet irradiation, and FK506 sequential resistance selection. FK506 production by the TCM8594 strain improved 45.1-fold ($505.4{\mu}g/mL$) compared to that of S. tsukubaensis NRRL 18488 ($11.2{\mu}g/mL$). Among the five substrates, wheat bran was selected as the best solid substrate to produce optimum quantities of FK506 ($382.7{\mu}g/g$ substrate) under solid-state fermentation, and the process parameters affecting FK506 production were optimized. Maximum FK506 yield ($897.4{\mu}g/g$ substrate) was achieved by optimizing process parameters, such as wheat bran with 5 % (w/w) dextrin and yeast extract as additional nutrients, 70 % (v/w) initial solid substrate moisture content, initial medium pH of 7.2, $30^{\circ}C$ incubation temperature, inoculum level that was 10 % (v/w) of the cell mass equivalent, and a 10 day incubation. The results showed an overall 234 % increase in FK506 production after optimizing the process parameters.

Selection of Protease Hyperproducing Mutant Strain from Serratia marcescens ATCC 2 1074 and Enzymatic Properties of the Protease (Serratia macescens ATCC 21074로부터 Protease 생산성이 높은 변이주의 선별 및 Protease의 효소학적 특성)

  • 김홍립;오평수
    • Microbiology and Biotechnology Letters
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    • v.19 no.5
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    • pp.450-455
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    • 1991
  • A protease hyperproducer, ampicillin resistant mutant, Serratia sp. SMNT-1 was selected from Serratia marcescens ATCC 21074 by mutagenesis. The protease productivity of this strain was about 11 times as much as that of the parental strain. The enzyme showed maximal activity at pH 9.0 and $40^{\circ}C$ and was stable over the pH range from 6.0 to 10.0 at $4^{\circ}C$, whereas it was unstable at $37^{\circ}C$ in alkaline condition. the enzyme was inactivated by heating at $60^{\circ}C$ for 10 min. The enzyme was inactivated by EDTA and reactivated by $Zn^{2+}, Co^{2+},\; and \; Mn^{2+}$, but the proteoiytic activity of the enzyme was not affected by DFP. From the above results, the protease produced by Serratia sp. SMNT-1 was classified as a metalloprotese.

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Isolation of Trp, Thr Overproducing Strain of Saccharomyces cerevisiae (Trp, Thr Analogue 복합 저항성 Saccharomyces cerevisiae 균주 개발)

  • 염형준;이승현;김선혜;선남규;안길환;이봉덕;원미선;송경빈
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.6
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    • pp.1017-1021
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    • 2004
  • To isolate a mutant which overproduces threonine and tryptophan, mutants of Saccharomyces cerevisiae were screened after UV and EMS mutagenesis. Hydroxynorvaline, a Thr analogue was used for selection of a Thr-overproducing mutant after UV mutagenesis. Among 31 mutants, TC 5-1 was selected as the strain candidate, based on amino acid analysis. TC 5-1 was then treated by EMS mutagenesis for Trp overproduction. Eight mutants were selected using fluorotryptophan for Thr and Trp overproducing strains. Amino acid analysis results showed that TC 6-1 was the best strain since it had the highest amount of Thr and Trp among mutants.

Structural damage identification based on genetically trained ANNs in beams

  • Li, Peng-Hui;Zhu, Hong-Ping;Luo, Hui;Weng, Shun
    • Smart Structures and Systems
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    • v.15 no.1
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    • pp.227-244
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    • 2015
  • This study develops a two stage procedure to identify the structural damage based on the optimized artificial neural networks. Initially, the modal strain energy index (MSEI) is established to extract the damaged elements and to reduce the computational time. Then the genetic algorithm (GA) and artificial neural networks (ANNs) are combined to detect the damage severity. The input of the network is modal strain energy index and the output is the flexural stiffness of the beam elements. The principal component analysis (PCA) is utilized to reduce the input variants of the neural network. By using the genetic algorithm to optimize the parameters, the ANNs can significantly improve the accuracy and convergence of the damage identification. The influence of noise on damage identification results is also studied. The simulation and experiment on beam structures shows that the adaptive parameter selection neural network can identify the damage location and severity of beam structures with high accuracy.