• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.72.2-73
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• 2003

We found a soybean (Glycine max) cultivar 561 that was strongly resistant to a virulent bacterial strain of a Pseudomonas spp. Further identification revealed that the Pseudomonas spp. was a strain of Pseudomonas aeruginosa. Furthermore we identified specific genes involved in the resistance of soybean 561 and analyzed the pattern of gene expression against the Pseudomonas infection using differential-display reverse transcription PCR (DDRT-PCR). More than 126 cDNA fragments representing mRNAs were induced within 48 hours of bacteria inoculation. Among them, 28 cDNA fragments were cloned and sequenced. Twelve differentially displayed clones with open reading frames had unknown functions. Sixteen selected cDNA clones were homologous to known genes in the other organisms. Some of the identified cDNAs were pathogenesis-related genes (PR genes) and PR-like genes. These cDNAs included a putative calmodulin-binding protein, an endo-1,3-1,4-b-D-glucanase, a b-1,3-endoglucanase, a b-1,3-exoglucanase, a phytochelatin synthetase-like gene, a thiol pretense, a cycloartenol synthase, and a putative receptor-like sorineithreonine protein kinase. Among them, we found that four genes were putative pathogenesis-related genes (PR) induced significantly by the p. aeruginosa infection. These included a calmodulin-binding protein gene, a b-1,3-endoglucanase gene, a receptor-like sorine/threonine protein kinase gene, and pS321 (unknown function). These results suggest that the differentially expressed genes may mediate the strong resistance of soybean 561 to Pseudomonas aeruoginosa.

• Horticultural Science & Technology
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• v.31 no.3
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• pp.275-281
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• 2013

Low temperature is one of the major environmental factors that affect productivity including reduced growth and budding of vines, and changes of metabolic processes in grape (Vitis spp.). To screen the specific expression of abiotic stress-related genes against cold treatment in 'Kyoho' and 'Campbell Early' grapevines, expression of various defense-related genes was investigated by RT-PCR and real-time PCR. Among the 67 genes analyzed by RT-PCR and real-time PCR, 17 and 16 types of cDNA were up-regulated, while 5 and 6 types were down-regulated in cold-treated 'Kyoho' and 'Campbell Early' grapevines, respectively. Genes encoding carotene (Cart3564 and Cart4472), chalcone isomerase (CHI), cytochrome P450 (CYP), flavonol synthase (FLS), endo-${\beta}$-glucanase precursor (Glu), glutathione peroxidase (GPX), glutathione-S-transferase (GST), leucine-rich repeats (LRR), manganese superoxide dismutase (Mn-SOD), phenylalanine ammonia lyase (PAL), polygalacturonase-inhibiting protein (PGIP), proline rich protein 2 (PRP2), small heat shock protein (sHSP), temperature induced lipocalin (TIL), and thaumatin-like protein (TLP) were up-regulated, while those encoding CBF like transcription factor (CBF1), chitinase-like protein (CLP), cold induced protein (CIP), glycerol-3-phosphate acyltransferase (GPAT), and mitogen-activated protein kinase (MAPK) were down-regulated by low temperature treatment in both in 'Kyoho' and 'Campbell Early'.

• Journal of the Korean Wood Science and Technology
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• v.38 no.2
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• pp.140-148
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• 2010

This study was to investigate the enzymatic hydrolysis of cellulose using the cellulase from whole body of the native termite collected in Milyang-si, Kyungsangnamdo, Korea. In the results, optimal temperature and pH for the enzyme of native termites were $45^{\circ}C$ and pH 5.5 for both endo-${\beta}$-1, 4-glucanase and ${\beta}$-glucosidase. Enzyme activity of the termite enzyme was shown $8.8{\times}10^{-2}\;FPU/m{\ell}$. And the highest glucose hydrolysis rate of cellulose by the digestive enzyme from test termites was 24.5% based on the glucan, comparing 59.7% by commercial enzyme (only celluclast 1.5 L) at 1% (w/v) substrate and 36 hours in hydrolysis time. This hydrolysis rate by the digestive enzyme from test termites was comparatively high value in 41% level of the commercial enzyme. When cellulose was hydrolyzed by the digestive enzyme of the native termite, glucose hydrolysis was almost completed in 12 hours which was the considerably reduced time for cellulose hydrolysis. It was suggested that the quiet short reaction time for cellulose hydrolysis by the enzyme from native termite could be a very high advantage for development of hydrolysis cellulase for lignocellulosic biomass.

• Journal of Microbiology and Biotechnology
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• v.16 no.4
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• pp.576-583
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• 2006

The alkali-soluble glucan of the yeast cell wall contains $\beta-(1,3)-$ and (1,6)-D-linkages and is known to systemically enhance the immune system. In the previous study [6], in order to isolate cell wall mutants, a wild-type strain was mutagenized by exposure to ultraviolet light, and the mutants were then selected via treatment with laminarinase $(endo-\beta-(1,3)-D-glucanase)$. The mass of alkali- and water-soluble glucans produced by the mutant was measured to be 33.8 mg/g of the dry mass of the yeast cell. Our results showed that the mutants generated the amount of alkali-soluble glucan 10-fold higher than that generated by the wild-type. Structural analysis showed that the alkali-soluble glucan from the mutants was associated with a higher degree of $\beta-(1,6)-D-linkage$ than was observed in conjunction with the wild-type. Yeast cell wall $\beta-glucan$ was shown to interact with macrophages via receptors, thereby inducing the release of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide. Alkali-soluble $\beta-glucans$, both from water-soluble and water-insoluble glucan, exhibited a higher degree of macrophage activity with regard to both the secretion of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide and direct phagocytosis, than did the positive control ($1{\mu}g$ of lipopolysaccharide).

• Journal of Microbiology and Biotechnology
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• v.24 no.9
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• pp.1196-1206
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• 2014

A metagenomic fosmid library was constructed using genomic DNA isolated from the gut microflora of Hermetia illucens, a black soldier fly. A cellulase-positive clone, with the CS10 gene, was identified by extensive Congo-red overlay screenings for cellulase activity from the fosmid library of 92,000 clones. The CS10 gene was composed of a 996 bp DNA sequence encoding the mature protein of 331 amino acids. The deduced amino acids of CS10 showed 72% sequence identity with the glycosyl hydrolase family 5 gene of Dysgonomonas mossii, displaying no significant sequence homology to already known cellulases. The purified CS10 protein presented a single band of cellulase activity with a molecular mass of approximately 40 kDa on the SDS-PAGE gel and zymogram. The purified CS10 protein exhibited optimal activity at $50^{\circ}C$ and pH 7.0, and the thermostability and pH stability of CS10 were preserved at the ranges of $20{\sim}50^{\circ}C$ and pH 4.0~10.0. CS10 exhibited little loss of cellulase activity against various chemical reagents such as 10% polar organic solvents, 1% non-ionic detergents, and 0.5 M denaturing agents. Moreover, the substrate specificity and the product patterns by thin-layer chromatography suggested that CS10 is an endo-${\beta}$-1,4-glucanase. From these biochemical properties of CS10, it is expected that the enzyme has the potential for application in industrial processes.

• The Plant Pathology Journal
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• v.19 no.5
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• pp.239-247
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• 2003

In Korea, a local soybean (Glycine max) genotype 56l. was found to be strongly resistant to a virulent bacterial strain of a Pseudomonas sp. SN239. Specific genes involved in the resistance of the soybean genotype 561 were identified and the pattern of gene expression against the Pseudomonas infection was analyzed using differential-display reverse transcription PCR (DDRT-PCR). More than 126 cDNA fragments representing mRNAs were induced within 48 hours of bacteria inoculation. Among them, 28 cDNA fragments were cloned and sequenced. Twelve differentially displayed clones with open reading frames had unknown functions. Sixteen selected cDNA clones were homologous to known genes of other organisms. Some of the identified cDNAs were pathogenesis-related (PR) genes and PR-like genes. These cDNAs included a putative calmodulin-binding protein; an endo-l,3-1,4-$\bate$-D-glucanase; a $\bate$-1,3-endoglucanase; a $\bate$-1,3-exoglucanase; a phytochelatin synthetase-like gene; a thiol protease; a cycloartenol synthase; and a putative receptor-like serine/threonine protein kinase. Among them, four genes were found to be putative PR genes induced significantly by the Pseudomonas infection. These included a calmodulin-binding protein gene, a $\bate$-1,3-endoglucanase gene, a receptor-like serine/threonine protein kinase gene, and pS321 (unknown function). These results suggest that the differentially expressed genes may mediate the strong resistance of soybean 561 to the strain SN239 of Pseudomonas sp.

• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.247-255
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• 2006

Yeast cell wall matrix particles are composed entirely of mannoprotein and ${\beta}-glucan$. The mannoproteins of yeast cell wall can systemically enhance the immune system. We previously purified and analyzed alkali-soluble ${\beta}-glucans$ [${\beta}$-(1,3)- and ${\beta}$-(1,6)-glucans] [10]. In the present study, a wild-type strain was first mutagenized with ultraviolet light, and the cell wall mutants were then selected by treatment with 1.0 mg/ml laminarinase (endo-${\beta}$-(1,3)-D-glucanase). Mannoproteins of Saccharomyces cerevisiae were released by laminarinase, purified by concanavalin-A affinity and ion-exchange chromatography. The results indicated that the mutants yielded 3-fold more mannoprotein than the wild-type. The mannoprotein mass of mutant K48L3 was 2.25 mg/100 mg of yeast cell dry mass. Carbohydrate analysis revealed that they contained mannose, glucose, and N-acetylglucosamine. Saccharomyces cerevisiae cell wall components, mannoproteins, are known to interact with macrophages through receptors, thereby inducing release of tumor necrosis factor alpha ($TNF-{\alpha}$) and nitric oxide. Mannoprotein tractions in the present study had a higher macrophage activity of secretion of $TNF-{\alpha}$ and nitric oxide and direct phagocytosis than positive control ($1{\mu}g$ of lipopolysaccharide). In particular, F1 and F3 fractions in mannoproteins of K48L3 enhanced and upregulated the activity of nitric oxide secretion and macrophage phagocytosis by approximately two- and four-fold, respectively.

• Journal of the Korean Wood Science and Technology
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• v.47 no.6
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• pp.692-699
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• 2019

Elizabethkingia miricola BM10, a symbiotic bacterium, has been isolated from the hindgut of Reticulitermes speratus KMT001, a termite which occurs on Bukhan Mountain in Seoul, Korea. This strain demonstrated a symbiotic characteristic, in that it lacked endo-${\beta}$-1,4-glucanase activity, in a previous study. The major fatty acids of E. miricola BM10 were iso-$C_{15:0}$, iso-$C_{17:0}$ 3-OH, and summed feature 3 (iso-$C_{16:1}{\omega}7c/C_{16:1}{\omega}6c$). The content of iso-$C_{17:0}$ 3-OH was higher, while those of ECL 13.566, iso-$C_{17:11}{\omega}9c$, and summed feature 4 were lower than the other three type-strains of the Elizabethkingia genus. The 16S rRNA phylogenetic analysis confirmed that E. miricola BM10 is a new species. The whole genome of E. miricola BM10 was sequenced. The average nucleotide identity of strain BM10 as evaluated by pairwise comparison with E. anophelis R26, E. meningoseptica ATCC 13253, and E. miricola GTC 862 was shown to be 91.5%, 81.2%, and 94.29%, respectively. Based on our study results, E. miricola BM10 appears to represent a new strain of the genus Elizabethkingia.

• Korean Journal of Microbiology
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• v.42 no.4
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• pp.319-325
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• 2006

[ $\beta$ ]-Glucans (AG) were prepared from Agaricus blazei cultured in the medium fortified with the roots of Pueraria spp. by repeated extraction with hot water, gel filtration chromatography and DEAE ion exchange chromatography. Oligosaccharides (AO) were derived from the hydrolysis of AG by an endo-$\beta$-(1$\rightarrow$6)-glucanase from Bacillus megaterium. The anti-HT-29 human colon cancer activity of AG or AO was investigated using MTT assay, apoptosis assay, cell cycle analysis, and cDNA microairay. AG and AO both inhibited proliferation and growth of HT-29 cells, and stimulated apoptosis of the cells in a dose-dependent manner. In cell cycle analysis, treating HT-29 cells with AG or AO resulted in the increase of cells in the G0 (sub-G1) and G1 phase. Especially, AO was more effective in inducing G0/G1 cell cycle arrest than AG. To screen the genes involved in the increase of apoptosis, the gene expression profile of the HT-29 cells treated with AO was examined by cDNA microarray. While several genes involved in cell cycle progression (CCND2 and CDK2) were down-regulated, many genes involved in apoptosis (TNFSF9, TNFRSF9, FADD, CASP8, BAD, CRADD, CASP9 etc), cell cycle inhibitor (CDKN2A), immune response (IL6, IL18, IL6R etc), and tumor suppressor (CEACAM1, TP53BP2, IRF1, and PHB) were up-regulated. These results suggest that AO could inhibit the proliferation and growth of HT-29 cells by G0/G1 cell cycle arrest and induction of apoptosis.