• Title/Summary/Keyword: Strain-control

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Creation of an Ethanol-Tolerant Yeast Strain by Genome Reconstruction Based on Chromosome Splitting Technology

  • Park, A-Hwang;Sugiyama, Minetaka;Harashima, Satoshi;Kim, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.184-189
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    • 2012
  • We sought to breed an industrially useful yeast strain, specifically an ethanol-tolerant yeast strain that would be optimal for ethanol production, using a novel breeding method, called genome reconstruction, based on chromosome splitting technology. To induce genome reconstruction, Saccharomyces cerevisiae strain SH6310, which contains 31 chromosomes including 12 artificial mini-chromosomes, was continuously cultivated in YPD medium containing 6% to 10% ethanol for 33 days. The 12 mini-chromosomes can be randomly or specifically lost because they do not contain any genes that are essential under high-level ethanol conditions. The strains selected by inducing genome reconstruction grew about ten times more than SH6310 in 8% ethanol. To determine the effect of mini-chromosome loss on the ethanol tolerance phenotype, PCR and Southern hybridization were performed to detect the remaining mini-chromosomes. These analyses revealed the loss of mini-chromosomes no. 11 and no. 12. Mini-chromosome no. 11 contains ten genes (YKL225W, PAU16, YKL223W, YKL222C, MCH2, FRE2, COS9, SRY1, JEN1, URA1) and no. 12 contains fifteen genes (YHL050C, YKL050W-A, YHL049C, YHL048C-A, COS8, YHLComega1, ARN2, YHL046W-A, PAU13, YHL045W, YHL044W, ECM34, YHL042W, YHL041W, ARN1). We assumed that the loss of these genes resulted in the ethanol-tolerant phenotype and expect that this genome reconstruction method will be a feasible new alternative for strain improvement.

Inhibition of Verticillium Wilt in Cotton through the Application of Pseudomonas aeruginosa ZL6 Derived from Fermentation Residue of Kitchen Waste

  • Qiuhong Niu;Shengwei Lei;Guo Zhang;Guohan Wu;Zhuo Tian;Keyan Chen;Lin Zhang
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1040-1050
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    • 2024
  • To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42℃), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ߡNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMߡNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.

Validating a Xylose Regulator to Increase Polyhydroxybutyrate Production for Utilizing Mixed Sugars from Lignocellulosic Biomass Using Escherichia coli

  • Suk-Jin Oh;Hong-Ju Lee;Jeong Hyeon Hwang;Hyun Jin Kim;Nara-Shin;Sang-Ho Lee;Seung-Oh Seo;Shashi Kant Bhatia;Yung-Hun Yang
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.700-709
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    • 2024
  • Polyhydroxybutyrate (PHB) production from lignocellulosic biomass is economically beneficial. Because lignocellulosic biomass is a mixture rich in glucose and xylose, Escherichia coli, which prefers glucose, needs to overcome glucose repression for efficient biosugar use. To avoid glucose repression, here, we overexpressed a xylose regulator (xylR) in an E. coli strain expressing bktB, phaB, and phaC from Cupriavidus necator and evaluated the effect of xylR on PHB production. XylR overexpression increased xylose consumption from 0% to 46.53% and produced 4.45-fold more PHB than the control strain without xylR in a 1% sugar mixture of glucose and xylose (1:1). When the xylR-overexpressed strain was applied to sugars from lignocellulosic biomass, cell growth and PHB production of the strain showed a 4.7-fold increase from the control strain, yielding 2.58 ± 0.02 g/l PHB and 4.43 ± 0.28 g/l dry cell weight in a 1% hydrolysate mixture. XylR overexpression increased the expression of xylose operon genes by up to 1.7-fold. Moreover, the effect of xylR was substantially different in various E. coli strains. Overall, the results showed the effect of xylR overexpression on PHB production in a non-native PHB producer and the possible application of xylR for xylose utilization in E. coli.

Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice

  • Chung, Eu Jin;Hossain, Mohammad Tofajjal;Khan, Ajmal;Kim, Kyung Hyun;Jeon, Che Ok;Chung, Young Ryun
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.152-164
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    • 2015
  • Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.

Evaluation of Bio-Control Efficacy of Trichoderma Strains against Alternaria alternata Causing Leaf Blight of Ashwagandha [Withania somnifera (L.) Dunal]

  • Rahman, Md. Ahsanur;Rahman, Md. Arifur;Moni, Zakiah Rahman;Rahman, Mohammad Anisur
    • Journal of Forest and Environmental Science
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    • v.36 no.3
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    • pp.207-218
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    • 2020
  • Ashwagandha is an important ancient medicinal crops, being affected with many diseases, among which leaf blight caused by Alternaria alternata has become the constraint resulting in huge yield losses. Continuous usage of chemical methods leads to environment, soil and water pollution. Whereas biological control of diseases is long lasting, inexpensive, eco-friendly and harmless to target organisms. In this context, it is aimed to evaluate five Trichoderma strains viz. Trichoderma virens IMI-392430, T. pseudokoningii IMI-392431, T. harzianum IMI-392432, T. harzianum IMI-392433 and T. harzianum IMI-392434 as bio-control efficacy against A. alternata and growth promoting effect in Ashwagandha. All the Trichoderma strains had varied antagonistic effects against the pathogen. In dual culture technique, the strain T. harzianum IMI-392433 showed maximum percentage inhibition of mycelial growth (54.89%) followed by T. harzianum IMI-392432 (53.83%), T. harzianum IMI-392434 (48.94%) and T. virens IMI-392430, (43.62%) against the pathogen, while the least inhibition percentage was observed with the T. pseudokoningii IMI-392431 (36.60%). The culture filtrate of the Trichoderma strain, T. harzianum IMI-392433 recorded highest inhibition on the mycelial growth (39.05%) and spore germination (80.77%) of pathogen and the lowest was recorded in T. pseudokoningii IMI-392431 (20.45 and 50%). Moreover, seeds treated with spore suspension of the strain T. harzianum IMI-392433 reduced the percentages of disease severity index significantly. The strain T. harzianum IMI-392433 also significantly increased seed germination %, seedling vigor and growth of Ashwagandha. The correlation matrix showed that root yield per plant of Ashwagandha had significant and positive correlation with plant height (r=0.726⁎⁎), number of leaf (r=0.514⁎⁎), number of primary branch (r=0.820⁎⁎), number of secondary branch (r=0.829⁎⁎), fresh plant weight (r=0.887⁎⁎), plant dry weight (r=0.613⁎⁎), root length (r=0.824⁎⁎), root diameter (r=0.786⁎⁎), root dry weight (r=0.739⁎⁎) and fresh root weight (r=0.731⁎⁎). The significant and negative correlation (r=-0.336⁎⁎) was observed with the root yield and percentages of disease severity index. The study recognized that the T. harzianum IMI-392433 strain performed well in inhibiting the mycelial growth and reduced the percentages of disease severity index of pathogen as well as increased the plant growth in Ashwagandha.

Pathogenicity of Korean Entomopathogenic Nematodes to Exomala orientalis(Coleoptera : Scarabaeidae) (등얼룩풍뎅이(Exomala orientalis)에 대한 한국산 곤충병원성 선충의 병원성)

  • Lee Dong-Woon;Kim Hyeons-Hwan;Lee Sang-Myeong;Choo Ho-Yul;Choi Woo-Goun;Kweon Tae-Woong
    • Asian Journal of Turfgrass Science
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    • v.19 no.1
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    • pp.39-46
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    • 2005
  • The 21 strains of Korean entomopathogenic nematodes, {Heterorhabditis bacteriophora Hamyang strain(HbH), Heterorhabditis sp. 202, 205, 217, Heterorhabdiris sp. KCTC 0991BP strain, Steinernema carpocapsae Pocheon(ScP), S. longicaudum Gonaju, S. longicaudum Nonsan, Steinernema sp. 7,24, 52, 55, 60, 64, 206, 207, 209, 210, 219, and 227 strain} were evaluated for the control of a turfgrass insect pest, Exomala orientalis. Heterorhabditis spp. showed higher pathogenicity than Steinernema spp. against 3rd instar larvae of E. orientalis with $55\%$ mortality by Heterorhabditis sp. 202 strain and $50\%$ by HbH and Heterorhabditis sp.205 strain at the rate of 200 infective juveniles per larva 14 days later after treatment. The number of infective juveniles of Korean entomopathogenic nematodes in 3rd instar larvae of E. orientalis was higher in Heterorhabditis spp. than in Steinernema spp.. In general, numbers of produced infective juveniles of three species were much higher, i.e., Heterorhabditis sp.202 strain produced 273,064 infective juveniles, S. carpocapsae Pocheon strain 273,043, and Heterorhabditis sp. 217 strain 248,887, respectively.

A Study on the Contact Seam Tracking Sensor by Using Strain Gauges (스트레인 게이지를 이용한 접촉식 용접선 추적 센서에 관한 연구)

  • 안병원;배철오;김현수
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.7 no.5
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    • pp.1019-1025
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    • 2003
  • There are many kinds of seam trackers in the industrial welding field. We are proposed the contact seam tracking sensor applying strain gauges kind of contact sensor that mostly used in welding part now. For this seam tracking experiment, we made the strain gauges sensor by ourselves and tested how well the sensor tracks the seam. The experiment device consist of strain gauges sensor, amplifier circuit of strain gauges signal, saw wave generator, MOSFET power diving circuit and X-Y slide by moved DC motor. The tracking areas are X-Y planes(left, right, up and down) and the change of strain gauge resistance causes to move DC motor that connected to X-Y slide. As a result of experiment, we confirmed that the strain gauges sensor tracks a seam well, and X-Y slide DC motor was controlled by PWM control.

A study on the thermal-mechanical fatigue life prediction of 12 Cr steel (12 Cr 강의 열피로 수명단축에 관한 연구)

  • Ha, Jeong-Soo;Kim, Kun-Young;Ahn, Hye-Thon
    • Journal of the Korean Society for Precision Engineering
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    • v.11 no.4
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    • pp.114-125
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    • 1994
  • Fatigue behavior and life prediction method were presented for themal-mechanical and isothermal low cycle fatigue of 12 Cr forged steel used for high temperature applications. In-phase and out-of-phase thermal-mechanical fatigue test from 350 .deg. C to 600 .deg. C and isothermal low cycle fatigue test at 600 .deg. C, 475 .deg. C, 350 .deg. C were conducted using smooth cylindrical hollow specimen under strain-control with total strain ranges from 0.006 to 0.015. The phase difference between temperature and strain in thermal-mechanical fatigue resulted in significantly shorter fatigue life for out-of-phase than for in-phase. Thermal-mechanical fatigue life predication was made by partitioning the strain ranges of the hysteresis loops and the results of isothermal low cycle fatigue tests which were performed under the combination of slow and fast strain rates. Predicted fatigue lives for out-of-phase using the strain range partitioning method showed an excellent agreement with the actual out-of-phase thermal-mechanical fatigue lives within a factor of 1.5. Conventional strain range partitioning method exhibited a poor accuracy in the prediction of in-phase range partitioning method in a conservative way. By the way life prediction of thermal-mechanical fatigue by Taira's equivalent temperature method and spanning fartor method showed good agreement within out-of-phase thermal-mechanical fatigue.

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Development of Thermal Distortion Analysis Method Based on Inherent Strain for TMCP Steels (TMCP 강판의 고유변형도 기반 열변형 해석법 개발)

  • Ha, Yun-Sok;Yang, Jin-Hyuk;Won, Seok-Hee;Yi, Myung-Su
    • Journal of Welding and Joining
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    • v.26 no.3
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    • pp.61-66
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    • 2008
  • As ships become to be larger than ever, the thicker plate and the higher tensile steel plate are used in naval shipyard. Though special chemical composition is needed for high-tensile steels, recent high-tensile steels are made by the TMCP(Thermo-Mechanical control process) skill. The increase of yield stress and tensile stress of TMCP steels is induced from bainite phase which is transformed from austenite, but that increased yield stress can be vanished by another additional thermal cycle like welding and heating. As thermal deformations are deeply related by yield stress of material, the study for prediction of plate deformation by heating should reflect principle of TMCP steels. This study developed an algorithm which can calculate inherent strain. In this algorithm, not only the mechanical principles of thermal deformations, but also the predicting of the portion of initial bainite is considered when calculating inherent strain. The simulations of plate deformation by these values showed good agreements with experimental results of normalizing steels and TMCP steels in welding and heating. Finally we made an inherent strain database of steels used in Class rule.

Low cycle fatigue behaviour of TMCP steel in as-received and welded states (TMCP 고장력강재와 그 용접부의 저사이클피로특성에 관한 연구)

  • 김영식;한명수
    • Journal of Welding and Joining
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    • v.8 no.4
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    • pp.46-57
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    • 1990
  • TMCP steel manufactured by controlled rolling followed by accelerated cooling process is known to have extra-ordinary mechanical properties such as tensile strength and toughness. However, there is much uncertainty about the fatigue fracture characteristics, especially, in the welded state of this steel. In case of this steel, the softening zone by welding is generated in heat affected zone in contrast with the case of conventional normalized high strength steel. This softening zone is considered to play significant roles in low cycle fatigue fracture of the welded part of this steel. In this paper, the low cycle fatigue behaviors of TMCP steel were inspected in as-received and welded state using the smooth specimen. The fatigue life-time was seperately investigated on the basis of failure of the specimen and crack initiation which is detected by differential strain method. Moreover, the low cycle fatigue characteristics of TMCP steel were quantitatively compared with those of the conventional normalized steel of same strength level.

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