• Transactions of the Korean Society of Mechanical Engineers A
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• v.28 no.8 s.227
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• pp.1183-1189
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• 2004

A novel cell count method, which can improve the count efficiency and reduce contamination problem, was presented using micro lattice engraved on culture dish. The micro lattice has feature of $50{\mu}m{\times}50{\mu}m$ rectangular shape, $2{\mu}m$ line width, and $2{\mu}m$ depth in $3mm{\times}3mm$ area. In this paper, nickel mold was fabricated with thickness of 3mm and diameter of 80 mm, and transcription of the micro lattice on a polystyrene cell culture dish was performed by hot embossing at $200\;^{\circ}C$. The tedious and error-prone harvest/load processes of conventional cell counts with a hemocytometer could be omitted, and these advantages became magnified during periodical counts involving long-term cultures. SupT1 cells and HeLa cells were cultivated with the dish for 7 days in $CO_2$ incubator and counted as $371.84/mm^2$ and $123.36/mm^2$, respectively, during the cultivation without harmful effects on the cells.

• Journal of Microbiology
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• v.38 no.1
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• pp.36-39
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• 2000

Toxic hexavalent chromium, Cr(VI), was reduced to a less toxic trivalent chromium form by E. coli ATCC 33456. The suitable electron donor for Cr(VI) reduction was glucose. E. coli ATCC 33456 was more resistant to metal cations than other reported Cr(VI) reducing microorganisms. Cell growth was inhibited by the presence of Cr(VI) in a liquid medium and Cr(VI) reduction accompanied cell growth. With a hydraulic retention time of 20 h, Cr(VI) reducing efficiency was 100% to 84% when Cr(VI) concentration in the influent was in the range of 10 to 40 mg L$\^$-1/. Specific rate of Cr(VI) reduction was 2.41 mg Cr(VI) g DCW$\^$-1/ h$\^$-1/ when 40 mg L$\^$-1/ of Cr(VI) influent was used. This result suggested the potential application of E. coli ATCC 33456 for the detoxification of Cr(VI) in Cr(VI) contaminated wastewater.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.45 no.6
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• pp.600-605
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• 2012

In this study, we investigated the mechanism of inhibition of pathogenic bacteria by Pediococcus pentosaceus strain SH-10 isolated from hard Clam Meretrix meretrix sikhae. When P. pentosaceus SH-10 was co-cultured in MRS broth with pathogenic bacteria, including Bacillus cereus, Listeria monocytogenes, Salmonella choleraesuis and Staphyloccus aureus, no viable pathogenic cells were detected after 18 h of incubation. However, pediocin or a pediocin-like bacteriocin was not detected in cultures of P. pentosaceus SH-10 by the agar diffusion method. Organic acids were produced in MRS broth in proportion to the incubation time of P. pentosaceus SH-10. These results indicate that P. pentosaceus SH-10 inhibited the growth of pathogenic bacteria by lowering the pH of the growth medium through the production of organic acids, including sodium lactate, sodium acetate, and sodium citrate.

• Development and Reproduction
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• v.17 no.1
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• pp.1-8
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• 2013

Osteoprotegerin (OPG) is a secreted glycoprotein that regulates bone resorption by inhibiting differentiation and activation of osteoclast, thereby potentially useful for the treatment of many bone diseases associated with increased bone loss. In this study, we designed a novel cDNA expression cassette by modifying the potent and mammary gland-specific goat ${\beta}$-casein/hGH hybrid gene construct and examined human OPG (hOPG) cDNA expression in transgenic mice. Six transgenic mice all successfully expressed hOPG in their milk at the level of 0.06-2,000 ${\mu}g/ml$. An estimated molecular weight of the milk hOPG was 55 kDa in SDS-PAGE, which is the same as a naturally glycosylated monomer. This hOPG expression was highly specific to the mammary glands of transgenic mice. hOPG mRNA was not detected in any organs analyzed except mammary gland. Functional integrity of milk hOPG was evaluated by TRAP (tartrate-resistant acid phosphatase) activity assay in bone marrow cell cultures. OPG ligand (OPG-L) treatment increased TRAP activity by two fold but it was completely abolished by co-treatment with transgenic milk containing hOPG. Taken together, our novel cDNA expression cassette could direct an efficient expression of biologically active hOPG, a potential candidate pharmaceutical for bone diseases, only in the mammary gland of transgenic mice.

• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1331-1337
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• 2006

A mathematical model has been proposed for the batch culture of Agaricus blazei with mixed carbon sources of glucose and dextrin. In the proposed model, the metabolism of A. blazei was divided into three parts: cell growth, exopolysaccharides (EPS) production, and another EPS production pathway activated by dextrin hydrolysis. Each pathway was described mathematically and incorporated into the mechanistic model structure. Batch cultures were carried out with six different carbon source compositions. Although parameters were estimated by using the experimental data from the two extreme cases such as glucose only and dextrin only, the model represented well the profiles of glucose, cell mass, and EPS concentrations for all the six different carbon source mixtures, showing a good interpolation capability. Of note, the lag in EPS production could be quite precisely predicted by assuming that the glucose-to-cell mass ratio was the governing factor for EPS production.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.518-521
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• 1999

Bacillus sp. 79-23 spores were irradiated with $^{60}Co$ gamma-rays at doses ranging from 0.5 to 5 kGy. Following gamma-irradiation, seven mutant strains were isolated by scoring the halo sizes formed around the colonies grown on LB agar plates containing 4% carboxymethylcellulose (CMC) and trypan blue. The mutant strains showed a 1.5 to 2-fold increase in carboxymethylcellulase (CMCase) activity over the parent strain. Wheat bran acted as an effective inducer for CMCase production in the parent and mutant strains. Mutant strains 68 and 70 were identified as exhibiting higher CMCase activities than those of other mutants in LB media both with and without 3% wheat bran. In addition, these strains seem to produce substantially lower amounts of capsular materials, whereas the parent strain produced large amounts of them in both liquid and solid LB media. In flask cultures, the CMCase production by mutants 68 and 70 reached maximum levels of 17.5 unit/ml and 15.7 unit/ml, respectively, in an LB medium containing 3% wheat bran.

• Journal of Microbiology
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• v.33 no.4
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• pp.295-301
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• 1995

An antifungal Pseudomonas stutzeri YPL-1 produced extracellular chitinase and .betha.-1, 3-glucanase that were key enzymes in the decomposition of fungal hyphal walls. These lytic extracellular enzymes markedly inhibited mycelial growth of the phytopathogenic fungus Fusarium solani. A chitinase from P. stutzeri YPL-1 inhibited fungal mycelial growth by 87%, whereas a .betha.-1, 3-glucanase from the bacterium inhibited growth by 53%. Furthermore, co-operative action of the enzymes synergistically inhibited 95% of the fungal growth. The lytic enzymes caused absnormal swelling and retreating on the fungal hyphal walls in a dual cultures. Scanning electron microscopy clearly showed hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. In an in vivo pot test, P. stutzeri YPL-1 proved to have biocontrol ability as a powerful agent in controlling plant disease. Planting of kidney bean (Phaseolus vulgaris L.) seedlings with the bacterial suspension in F. solani-infested soil significantly suppressed the development of fusarial root-rot. The characteristics of a crude preparation of .betha.-1, 3-glucanase produced from P. stutzeri YPL-1 were investigated. The bacterium detected after 2 hr of incubation. The enzyme had optimum temperature and pH of 40.deg.C and pH 5.5, respectively. The enzyme was stable in the pH range of 4.5 to 7.0 and at temperatures below 40.deg.C, with a half-life of 40 min at 60.deg.C.

• Molecules and Cells
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• v.37 no.3
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• pp.248-256
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• 2014

N-acetylglucosamine kinase (GlcNAc kinase or NAGK; EC 2.7.1.59) is a N-acetylhexosamine kinase that belong to the sugar kinase/heat shock protein 70/actin superfamily. In this study, we investigated both the expression and function of NAGK in neurons. Immunohistochemistry of rat brain sections showed that NAGK was expressed at high levels in neurons but at low levels in astrocytes. Immunocytochemistry of rat hippocampal dissociate cultures confirmed these findings and showed that NAGK was also expressed at low levels in oligodendrocytes. Furthermore, several NAGK clusters were observed in the nucleoplasm of both neuron and glia. The overexpression of EGFP- or RFP (DsRed2)-tagged NAGK in rat hippocampal neurons (DIV 5-9) increased the complexity of dendritic architecture by increasing the numbers of primary dendrites and dendritic branches. In contrast, knockdown of NAGK by shRNA resulted in dendrite degeneration, and this was prevented by the co-expression of RFP-tagged NAGK. These results suggest that the upregulation of dendritic complexity is a non-canonical function of NAGK.

• Journal of Microbiology and Biotechnology
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• v.31 no.3
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• pp.447-455
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• 2021

Strains of four Bacillus spp. were respectively inoculated into sterilized soybeans and the free amino acid profiles of the resulting cultures were analyzed to discern their metabolic traits. After 30 days of culture, B. licheniformis showed the highest production of serine, threonine, and glutamic acid; B. subtilis exhibited the highest production of alanine, asparagine, glycine, leucine, proline, tryptophan, and lysine. B. velezensis increased the γ-aminobutyric acid (GABA) concentration to >200% of that in the control samples. B. sonorensis produced a somewhat similar amino acid profile with B. licheniformis. Comparative genomic analysis of the four Bacillus strains and the genetic profiles of the produced free amino acids revealed that genes involved in glutamate and arginine metabolism were not common to the four strains. The genes gadA/B (encoding a glutamate decarboxylase), rocE (amino acid permease), and puuD (γ-glutamyl-γ-aminobutyrate hydrolase) determined GABA production, and their presence was species-specific. Taken together, B. licheniformis and B. velezensis were respectively shown to have high potential to increase concentrations of glutamic acid and GABA, while B. subtilis has the ability to increase essential amino acid concentrations in fermented soybean foods.

• 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.