• Korean Journal of Microbiology
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• v.25 no.1
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• pp.46-51
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• 1987

In order to increase the production of tryptophan by maximizing expression of recombinant trp plasmid, Klebsiella pneumoniae KC 105(pheA tyrA trpE trpR tyrR) was genetically modified. KC 107, inosine monophospate(IMP) auxotroph from KC 105 and KC 108, histidine(His) auxotroph from KC 107 were also derived respectively to increase phosphoribosylpyrophosphate(PRPP) production which is required for tryptophan biosynthesis. From KC 107 phosphoribosylpyrophosphate consumption which is required for tryptophan biosynthesis. From KC 107 and KC 108, KC 109 and KC 110, both arginine auxotrophs were derived respectively. To investigate the expression of recombinant trp plasmid in the selected K. pneumoniae mutants, the auxotrophic mutants were transformed with recombinant trp plasmids pSC 101-$trpE^{FBR}$, pSC 101-trpL(.DELTA.att) $trpE^{FBR}$ (pSC 101-trp-AF). Amount of tryptophan produced and activities of tryptophan synthase of $trpR^{-}$ mutant (KC 100) and $tyrR^{-}$ mutnat(KC 105) containing recombinant plasmid pSC 101-trp operon were increased by 30-40% as compared with KC 99(pheA tyrA trpE) containing recombinant plasmid pSC 101-trp operon. Activities of tryptophan synthase and production of tryptophan of KC 108 ($His^{-}$) and KC 109($Arg^{-}$) containing recombinant plasmid pSC 101-trp operon were increase by two-fold as compared with KC 107 containing pSC 101-trp operon.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.1
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• pp.11-14
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• 1998

A toluene-oxidizing strain of Pseudomanas mendocina KR1 containing toluene-4-mono-oxygenase (TMO) completely degrades TCE with the addition of toluene as a co-substrate in aerobic condition. In order to construct in situ bioremediation system for TCE degradation without any growth-stimulating nutrients or toxic inducer such as toluene, we used the carbon-starvation promoter of Pseudomonas putida MK1 (Kim, Y. et al., J. bacteriol., 1995). Upon entry into the stationary phase due to the deprivation of nutrients, this promoter is strongly induced without further cell growth. The TMO gene cluster (4.5 kb) was spliced downstream of the carbon starvation promoter of Pseudomonas putida MK1, already cloned in pUC19. TMO under the carbon starvation promoter was not expressed in E. coli cells either in stationary phase or exponential phase. For TMO expression in Pseudomonas strains, tmo and carbon starvation promoter region were recloned into a modified broad-host range vector pMMB67HES which was made from pMMB67HE(8.9 kb) by deletion of tac promoter and lacIq (about 1.5 kb). Indigo was produced by TMO under the carbon starvation promoter in a Pseudomonas strain of post-exponential phase on M9 (0.2% glucose and 1mM indole) or LB. 18% of TCE was degraded in 14 hours after entering the stationary phase at the initial concentration of 6.6 ${\mu}$M in liquid phase.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.774-783
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• 2007

In the present study, acidocin 1B, a bacteriocin produced by Lactobacillus acidophilus GP 1B, exhibited profound inhibitory activity against a variety of LAB and pathogens, including Gram-negative bacteria, and its mode of action was to destabilize the cell wall, thereby resulting in bactericidal lysis. Acidocin 1B was found to be heat stable, because it lost no activity when it was heated up to $95^{\circ}C$ for 60 min. It retained approximately 67% of the initial activity after storage for 30 days at $4^{\circ}C$, and 50% of its initial activity after 30 days at $25^{\circ}C$ and $37^{\circ}C$. The molecular mass of acidocin 1B was estimated to be 4,214.65 Da by mass spectrometry. Plasmid curing results indicated that a plasmid, designated as pLA1B, seemed to be responsible for both acidocin 1B production and host immunity, and that the pLA1B could be transformed into competent cells of L. acidophilus ATCC 43121 by electroporation. Our findings indicate that the acidocin 1B and its producer strain may have potential value as a biopreservative in food systems.

• International Journal of Industrial Entomology and Biomaterials
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• v.32 no.2
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• pp.49-53
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• 2016

Bone morphogenetic proteins (BMPs) are essential growth factors for bone formation, skeletal development and bone regeneration. The BMP-2/7 heterodimer is known to have remarkable effects on osteogenic induction that are even stronger than the BMP-2 or BMP-7 homodimers. We designed a recombinant human BMP-2/7 (rhBMP-2/7) heterodimer protein with four glycine residues between BMP-2 and BMP-7 protein to facilitate free bond rotation of domains. The Baculovirus Expression Vector System (BEVS) is routinely used to produce recombinant proteins in the milligram scale. In this study, the BEVS was used to express the rhBMP-2/7 protein whrer the recombinant baculovirus was recovered in the host Sf9 cells. To confirm the biological activity of rhBMP-2/7 protein secreted from the BEVS as an osteogenic differentiation and induction factor, we measured the BMP-induced ALP activity. rhBMP-2/7 could be used as an alternative to BMPs to overcome limitations like short half-life and requirement for high concentrations. Furthermore, rhBMP-2/7 may be an efficient tool for various application studies such as bone regeneration and skeletal development.

• Korean Journal of Veterinary Research
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• v.33 no.4
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• pp.633-647
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• 1993

NC strain mice were inoculated intraperitoneally with Dermatophoilus-like microorganism strain W254 isolated from the sulfur granules of the swine tonsils. The development of the resulting abscesses in NC strain mice was investigated at regular time intervals by means of histological, histochemical and electron microscopical methods. The results obtained were as follows ; 1. The inoculated bacteria have settled in the subserous site of the peritoneal organ and provoked cellular response to form neutrophil, macrophage and connective tissue layers at the time of one week after inoculation, and thereafter were found to have formed typical actinomycotic abscess two weeks after inoculation. 2. The persistant abscesses were appeared to reveal close interaction between higher activity of organism and tissue reaction and contained hyaloid bodies within or outside bacterial clumps. 3. The hyaline materials were appeared to be composed of protein rich in hydroxyphenyl group by histochemical examination. The materials were presumed to be produced by the interaction between bacterial activity and host cell reaction. 4. The bacteria in the abscesses showed the various developmental stages of coccoid body, mulberry-like body tuber-shaped body and filamentous body, and it was confirmed that the mesosomal development was related to septation of the bacterial cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.1
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• pp.74-80
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• 2012

Candida albicans is a common opportunistic pathogen and is frequently associated with biofilm formation occurring on the surfaces of host tissues and medical devices. On account of the distinct resistance of C. albicans biofilms to the conventional antifungal agents, new strategies are required to cope with these infections. The root of Polygonum cuspidatum has been used for medicinal purposes in East Asia. The aim of this study was to assess the anticandidal potential of the P. cuspidatum ethanol extract by evaluating biofilm formation, integrity of the cell membranes of C. albicans and adhesion of C. albicans cells to polystyrene surfaces. The growth and development of the biofilm was assessed using an XTT reduction assay, and the extract (0.39 mg/ml) significantly reduced ($41.1{\pm}17.8%$) biofilm formation of 11 C. albicans strains. The extract damaged the cell membranes of C. albicans and remarkably inhibited cell adhesion to polystyrene surfaces. The plant extract displayed fungistatic activity without significant hemolytic activity. Based on the results of this study, the P. cuspidatum extract has promising potential for use in treating biofilm-associated Candida infection.

• Development and Reproduction
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• v.24 no.4
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• pp.277-285
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• 2020

The disease-causing koi herpes virus (KHV), also known as cyprinid herpesvirus-3 (CyHV3), causes mass mortality of koi and carp. Koi (Cyprinus carpio) is a host for KHV, one of 12 virus species in the Alloherpesviridae family. We examined the effects of KHV disease koi (KK), and on koi×red common carp (KR) and red common carp×koi (RK) cross, using a virus challenge test. The infected fish had clinical signs that included gill necrosis and skin lesions. The RK and KR were highly more resistant (cumulative mortality: RK; 6% and KR; 8%) to KHV infection than KK fish (cumulative mortality: 28%). KHV DNA was confirmed in the tissues of all dead fish in groups by use of polymerase chain reaction (PCR), and the presence of the KHV protein in kidney was confirmed by immunohistochemistry. Histological analysis showed severe gill lesions and fusion of the lamellae in KK fish, but less severe damage in RK fish. In immunohistochemistry analysis, the KHV protein localized in the cytoplasm of infected kidney cells of KK, but the cross groups had lower levels of KHV antigen. Our data indicate that the cross groups had increased resistance to KHV disease.

• Parasites, Hosts and Diseases
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• v.57 no.3
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• pp.295-298
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• 2019

Fasciolopsiasis is rarely known as the parasitic disease in Nepal. Herein, we report a case of fasciolopsiasis in a 22-year-old man who was admitted in the hospital with abdominal pain, distension and loss of appetite for a month. He had previously diagnosed with acute viral hepatitis but, his abdominal pain was not resolving despite improvement in his liver function and general condition. During endoscopy an adult digenean worm was seen in the first part of the duodenum. After isolation, the worm was identified morphologically as Fasciolopsis buski. Microscogic examination of the patient's stool revealed eggs with a morphology consistent with F. buski. Eggs were yellow-brown, ellipsoidal, unembmbryonated, operculated, filled with yolk cells, with thin shell and ranging $118-130{\mu}m$ in length and $60-69{\mu}m$ in width. The abdominal pain of the patient was resolved after treatment with praziquantel. By the present study, it was confirmed for the first time that fasciolopsiasis is indigenously transmitted in Nepal. Accordingly, the epidemiological studies in humans and reservoir host animals should be performed intensively in near future.

• Microbiology and Biotechnology Letters
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• v.47 no.2
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• pp.242-249
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• 2019

Biosynthesis of indole-3-acetate (IAA) from L-tryptophan via indole-3-pyruvate pathway requires three enzymes including aminotransferase, indole-3-pyruvate decarboxylase, and indole-3-acetate dehydrogenase. To establish a bio-based production of IAA, the aspC, ipdC, and iad1 from Escherichia coli, Enterobacter cloacae, and Ustilago maydis, respectively, were expressed under control of the tac, ilvC, and sod promoters in C. glutamicum. Cells harboring ipdC produced tryptophol, indicating that the ipdC product is functional in this host. Analyses of SDS-PAGE and enzyme activity revealed that genes encoding AspC and Iad1 were efficiently expressed from the sod promoter, and their enzyme activities were 5.8 and 168.5 nmol/min/mg-protein, respectively. The final resulting strain expressing aspC, ipdC, and iad1 produced 2.3 g/l and 7.3 g/l of IAA from 10 g/l L-tryptophan, respectively, in flask cultures and a 5-L bioreactor.

• Journal of Microbiology and Biotechnology
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• v.29 no.7
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• pp.1137-1143
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• 2019

Hepatitis E virus (HEV) accounts for 20 million infections in humans worldwide. In most cases, the infections are self-limiting while HEV genotype 1 infection cases may lead to lethal infections in pregnant women (~ 20% fatality). The lack of small animal models has hampered detailed analysis of virus-host interactions and HEV-induced pathology. Here, by employing a recently developed culture-adapted HEV, we demonstrated that methyltransferase, a non-structural protein, strongly inhibits melanoma differentiation-associated gene 5 (MDA5)-mediated activation of type I interferon responses. Compared to uninfected controls, HEV-infected cells display significantly lower levels of $IFN-{\beta}$ promoter activation when assessed by luciferase assay and RT-PCR. HEV genome-wide screening showed that HEV-encoded methyltransferase (MeT) strongly inhibits MDA5-mediated transcriptional activation of $IFN-{\beta}$ and $NF-{\kappa}B$ in a dose-responsive manner whether or not it is expressed in the presence/absence of a tag fused to it. Taken together, current studies clearly demonstrated that HEV MeT is a novel antagonist of MDA5-mediated induction of $IFN-{\beta}$ signaling.