• Journal of Microbiology and Biotechnology
• /
• v.25 no.2
• /
• pp.162-173
• /
• 2015

The cellular fatty acid composition is important for metabolic plasticity in Rhodobacter sphaeroides. We explored the effects of changing the cellular ratio of unsaturated fatty acids (UFAs) to saturated fatty acids (SFAs) in R. sphaeroides by overexpressing several key fatty acid biosynthetic enzymes through the use of expression plasmid pRK415. Bacteria containing the plasmid pRKfabI₁ with the fabI₁ gene that encodes enoyl-acyl carrier protein (ACP) reductase showed a reduction in the cellular UFA to SFA ratio from 4 (80% UFA) to 2 (65% UFA) and had decreased membrane fluidity and reduced cell growth. Additionally, the ratio of UFA to SFA of the chromatophore vesicles from pRKfabI₁-containing cells was similarly lowered, and the cell had decreased levels of light-harvesting complexes, but no change in intracytoplasmic membrane (ICM) content or photosynthetic (PS) gene expression. Both inhibition of enoyl-ACP reductase with diazaborine and addition of exogenous UFA restored membrane fluidity, cell growth, and the UFA to SFA ratio to wild-type levels in this strain. R. sphaeroides containing the pRKfabB plasmid with the fabB gene that encodes the enzyme β-ketoacyl-ACP synthase I exhibited an increased UFA to SFA ratio from 4 (80% UFA) to 9 (90% UFA), but showed no change in membrane fluidity or growth rate relative to control cells. Thus, membrane fluidity in R. sphaeroides remains fairly unchanged when membrane UFA levels are between 80% and 90%, whereas membrane fluidity, cell growth, and cellular composition are affected when UFA levels are below 80%.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.7
• /
• pp.1333-1340
• /
• 2016

The main objective of this study was to investigate whether Lactobacillus rhamnosus GG (LGG) ameliorated the effects of Citrobactor rodentium infection in Toll-like receptor 2 (TLR2) knockout (KO) and TLR4 KO mice, as well as in wild-type C57BL/6 (B6) mice. TLR2 KO, TLR4 KO, and B6 mice were divided into three groups per each strain. Each group had an uninfected control group (n = 5), C. rodentium-infected group (n = 8), and LGG-pretreated C. rodentium-infected group (n = 8). The survival rate of B6 mice infected with C. rodentium was higher when pretreated with LGG. Pretreatment with LGG ameliorated C. rodentium-induced mucosal hyperplasia in B6 and TLR4 KO mice. However, in C-rodentium-infected TLR2 KO mice, mucosal hyperplasia persisted, regardless of pretreatment with LGG. In addition, LGG-pretreated B6 and TLR4 KO mice showed a decrease in spleen weight and downregulation of tumor necrosis factor alpha, interferon gamma, and monocyte chemotactic protein 1 mRNA expression compared with the non-pretreated group. In contrast, such changes were not observed in TLR2 KO mice, regardless of pretreatment with LGG. From the above results, we conclude that pretreatment with LGG ameliorates C. rodentium-induced colitis in B6 and TLR4 KO mice, but not in TLR2 KO mice. Therefore, LGG protects mice from C. rodentium-induced colitis in a TLR2-dependent manner.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.10
• /
• pp.1001-1011
• /
• 2011

To explore bacterial diversity for elucidating genetic variability in acylhomoserine lactone (AHL) lactonase structure, we screened 800 bacterial strains. It revealed the presence of a quorum quenching (QQ) AHL-lactonase gene (aiiA) in 42 strains. These 42 strains were identified using rrs (16S rDNA) sequencing as Bacillus strains, predominantly B. cereus. An in silico restriction endonuclease (RE) digestion of 22 AHL lactonase gene (aiiA) sequences (from NCBI database) belonging to 9 different genera, along with 42 aiiA gene sequences from different Bacillus spp. (isolated here) with 14 type II REs, revealed distinct patterns of fragments (nucleotide length and order) with four REs; AluI, DpnII, RsaI, and Tru9I. Our study reflects on the biodiversity of aiiA among Bacillus species. Bacillus sp. strain MBG11 with polymorphism (115Alanine > Valine) may confer increased stability to AHL lactonase, and can be a potential candidate for heterologous expression and mass production. Microbes with ability to produce AHL-lactonases degrade quorum sensing signals such as AHL by opening of the lactone ring. The naturally occurring diversity of QQ molecules provides opportunities to use them for preventing bacterial infections, spoilage of food, and bioremediation.

• Mycobiology
• /
• v.32 no.2
• /
• pp.88-94
• /
• 2004

Protoplasts of the wild type strain of Pleurotus osteatus were mutagenized with UV light, and 3,000 colonies were examined for abnormal mycelial and fruiting phenotypes. Forty one strains displayed variant phenotypes in mycelia and fruiting processes. The variant phenotypes were classified into 6 groups: (1) auxotrophic strains, which are incapable of growing on minimal media and can only grow when provided with their specific requirements; (2) abnormal vegetative strains, which grow very slowly on minimal and complete media; (3) primordiumless strains, which fail to develop to the formation of primordia; (4) maturationless strains, which form primordia, but do not form mature fruiting bodies; (5) specifically colored strains, which have Specific bluish grey or bluish white pileus; (6) poorly spored strains, which fail to produce basidiospore or which produce few spores. These variant strains may be useful in genetic breeding programs and for the studies of fungal development and genetics.

• Korean Journal of Materials Research
• /
• v.20 no.7
• /
• pp.385-391
• /
• 2010

Twinning-induced plasticity (TWIP) steels have attracted great attention due to their excellent mechanical properties of high tensile strength (over 800MPa) and high ductility (over 50%), which result from the high strain hardening due to the mechanical twin formation during plastic deformation. The purpose of this study is to investigate the effect of annealing temperature and alloying elements on the mechanical properties of Fe-18Mn-0.6C TWIP steel. In 1.5%Al TWIP steel with 0.123%Ti content, the average recrystallized grain size was reduced to 2.5 ${\mu}m$ by cold rolling and annealing at $800^{\circ}C$ for 5 min, because of the pinning effect of the fine TiC carbides on grain coarsening. The tensile strength was decreased and the ductility was improved with the increase of the annealing temperature. However, a reversion of hardness and yield strength happened between $750^{\circ}C$ and $800^{\circ}C$ due to TiC and $M_3C$ type precipitation. 0.56% Ni added TWIP steel exhibited relatively lower yield strength, because Ni precipitates were not formed during the annealing process. When this specimen was annealed at $800^{\circ}C$ for 5min, the tensile strength and elongation were revealed at 1096MPa and 61.8%, respectively.

• Proceedings of the KWS Conference
• /
• 2002.10a
• /
• pp.583-588
• /
• 2002

The purpose of the study is to evaluate fracture toughness on the Laser and the electron beam welded joints of high tensile steels (HT500, HT550, HT650) by using 3-point bend CTOD and Charpy impact test. WM (weld metal) CTOD tests have been carried out using two kinds of CTOD specimen, the Laser beam welding (108mm length, and 24mm width, and 12mm thickness) and the electron beam welding (l71mm length, and 38mm width, and 19mm thickness). WM Charpy impact specimen is a standard V-notch type, and the temperature of the experiment is changed from -45 to 20 degree of centigrade. FE-analysis is also performed in order to investigate the effect of stress-strain fields on fracture characteristics. Results of the standard V-notch Charpy test are influenced by strength mis-match effect and the absorbed energy vE depends on crack path, and The transition temperature of Laser beam welded joints is more higher than that of electron beam welded joints. Results of the 3-point bend test give low critical CTOD and the crack path is in the weld metal of al specimens. These results indicate fracture toughness characteristics of the welded joints and transition temperature of HT500 are similar both a Laser beam welded joint and an electron beam welded joint. But the fracture toughness and the transition temperature of the electron beam welded joints of HT550 and HT650 are higher than those o the Laser beam welded joints.

• Korean Journal of Veterinary Research
• /
• v.38 no.2
• /
• pp.371-378
• /
• 1998

Five strains of gram-negative and yellow-pigmented bacteria were recently isolated from diseased freshwater fishes in Korea. All isolates were confirmed as a known fish pathogen of columnaris disease, Cytophaga columnaris based on their colonial and cellular morphology, and on physiological, biochemical and antigenic characteristics. Although the isolates were from different fish species, their characteristics of them were very similar to those of the reference strains of C columnaris (NCMB $2248^T$ and EK 28). Also, profiles of OMPs of Korean isolates were similar to those of the reference strain, C columnaris NCMB $2248^T$ when analyzed by SDS-PAGE.

• Journal of Wetlands Research
• /
• v.19 no.4
• /
• pp.443-450
• /
• 2017

The objective of this study was to isolate algae growth inhibiting microorganism to biologically control Microcystis aeruginosa, which is a harmful cyanobacterium. Various bacterial strains were isolated in this study, and four bacterial strains of M1~M4 exhibited remarkable growth inhibiting activity against M. aeruginosa. Based on the 16S rRNA analysis, the isolated M1~M4 strains were identified, and isolated four strains were rod-type and gram-negative. In particular, as well as respective single strain, co-culture of the isolated M1~M4 strains showed obvious algicidal activity against M. aeruginosa. When mixed four strains were inoculated, about 50% of the chlorophyll a was reduced after two days, about 70% after four days, and about 80% after seven days. From these results mentioned above, the four bacterial strains may contribute to the control of harmful M. aeruginosa.

• Bulletin of the Korean Chemical Society
• /
• v.28 no.12
• /
• pp.2303-2309
• /
• 2007

Expressed protein ligation (EPL) technique, joining recombinantly expressed proteins to polypeptides, has been widely adopted for addressing various biological questions and for drug discovery. However, joining two recombinant proteins together is sometimes difficult when proteins are expressed insoluble and unrefoldable, because ligation-active proteins via intein-fusion are obtainable when they are folded correctly. We overcame this limitation coexpressing target protein with additional methionine aminopeptidase (MAP) which enhances removal of the initiation methionine of recombinantly expressed protein. Our approach demonstrated that two domains of 46 kDa 5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase, a target of herbicide glyphosate, were successfully joined by native chemical ligation, although its C-terminal domain was expressed as an inclusion body. The intein-fused N-terminal fragment of EPSP synthase (EPSPSN, residues 1-237) was expressed and the ligation-active thioester tagged N-terminal fragment (EPSPSN-thioester) was purified using a chitin affinity chromatography and mercapto-ethanesulphonate (MESNA) as intein thiolysis reagent. Its Cterminal fragment (EPSPSC, residues Met237-238CYS-427), expressed as an inclusion body, was prepared from an additional MAP-expressing strain. Protein ligation was initiated by mixing ~1 mM of EPSPSN-thioester with ~2 mM of EPSPSCCYS (residues 238CYS-427). Also we found that addition of 2% thiophenol increased the ligation efficiency via thiol exchange. The ligation efficiency was ~85%. The ligated full-length EPSP synthase was dissolved in 6 M GdHCl and refolded. Circular dichroism (CD) and enzyme activity assay of the purified protein showed that the ligated enzyme has distinct secondary structure and ~115% specific activity compared to those of wild-type EPSP synthase. This work demonstrates rare example of EPL between two recombinantly expressed proteins and also provides hands-on protein engineering protocol for large proteins.

• Structural Engineering and Mechanics
• /
• v.24 no.1
• /
• pp.1-18
• /
• 2006

Strengthening of existing old structures has traditionally been accomplished by using conventional materials and techniques, viz., externally bonded steel plates, steel or concrete jackets, etc. Alternatively, fibre reinforced polymer composite (FRPC) products started being used to overcome problems associated with conventional materials in the mid 1950s because of their favourable engineering properties. Effectiveness of FRPC materials has been demonstrated through extensive experimental research throughout the world in the last two decades. However there is a need to use refined analytical tools to simulate response of strengthened system. In this paper, an attempt has been made to develop a numerical model of strengthened reinforced concrete (RC) beams with FRPC laminates. Material models for RC beams strengthened with FRPC laminates are described and verified through a nonlinear finite element (FE) commercial code, with the help of available experimental data. Three dimensional (3D) FE analysis has been performed by assuming perfect bonding between concrete and FRPC laminate. A parametric study has also been performed to examine effects of various parameters like fibre type, stirrup's spacing, etc. on the strengthening system. Through numerical simulation, it has been shown that it is possible to predict accurately the flexural response of RC beams strengthened with FRPC laminates by selecting an appropriate material constitutive model. Comparisons are made between the available experimental results in literature and FE analysis results obtained by the present investigators using load-deflection and load-strain plots as well as ultimate load of the strengthened beams. Furthermore, evaluation of crack patterns from FE analysis and experimental failure modes are discussed at the end.