• Korean Journal of Microbiology
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• v.25 no.4
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• pp.339-345
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• 1987

A bacterium K-4-3, producing $\beta$-glucan hydrolyzing enzyme, was isolated from soil and identified to be Bacillus subtilis by its morpholohical and physiological characteristics. $\beta$-glucan was coloured using cibacron blue 3G-A and cross linded by the addition of 1, 4-butanedioldiglycidyl ether. This substrate was used for the isolation of $\beta$-glucanase producing microorganism. The $\beta$-glucan hydrolyzing enzyme actibity from isolated K-4-3 strain was also measured using the modified substrate. Bacillus subtilis K-4-3 produced the highest extracellular $\beta$-glucan hydrolyzing activity in the basal medium containing $\beta$-glucan as a carbon source, peptone and tryptone as a nitrogen source, and magnesium sulfate as an inorganic salt. The optimum temperature and initial pH for $\beta$-glucanase production by Bacillus subtilis K-4-3 were $37^{\circ}C$ and pH6. The highest enzyme activity was obtained at the culture age of 54 hrs with rotary shaking at $37^{\circ}C$. The crude enzyme showed the highest activity at pH 7.5-8.0 and $65^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.23 no.6
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• pp.652-658
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• 1995

We attempted simultaneous expression of genes coding for endoglucanase and $\beta $-glucosidase from Pseudomonas sp. by using a synthetic two-cistron svstem in Escherichia coli and Saccharomyces cerevisiae. Two-cistron system, 5'--tac promoter-endoglucanase gene--$\beta $-glucosidase gene-- 3', 5'-tac promoter--$\beta $-glucosidase gene--endoglucanase gene--3' and 5'-tac promoter--endoglucanase gene--SD sequence--$\beta $-glucosidase gene--3, were constructed, and expressed in E. coli and S. cerevisiae. The E. coli and S. cerevisiae contained two-cistron system produced simultaneously endoglucanase and $\beta $-glucosidase. The recombinant genes contained the bacterial signal peptide sequence produced low level of endoglucanase and $\beta $-glucosidase in S. cerevisiae transformants: Approximately above 44% of two enzymes was localized in the intracellular fraction. The production of endoglucanase and $\beta $-glucosidase in veast was not repressed in the presence of glucose or cellobiose. The veast strain contained recombinant DNA with two genes hydrolyzed carboxvmethyl cellulose, and these endoglucanase and $\beta $-glucosidase degraded CMC synergistically to glucose, cellobiose and oligosaccharide. This result suggests the possibility of the direct bioconversion of cellulose to ethanol by the recombinant yeast.

• YAKHAK HOEJI
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• v.54 no.6
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• pp.441-448
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• 2010

In present study, classification and quality control of Genus Polygonatum were developed using the isolated from Polygonati Odorati Rhizoma and Polygonati Rhizoma. 3 components were isolated from Butanol fractions of Polygonati Rhizoma, and 2 components were isolated from Hexane and Butanol fractions of Polygonati Odorati Rhizoma. All the components were obtained using silica gel and ODS column chromatography. The compounds were identified as adenosine, 14-hydroxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-D-galactopyranosyl-26-O-${\beta}$-D-glucopyranoside, 22-O-methyl-14-hydrocxyfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-Dgalactopyranosyl-26-O-${\beta}$-D-glucopyranoside, ${\beta}$-Sitosteryl-3-O-${\beta}$-D-D-glucopyranoside, 14-hydoxylfurost-5-ene-3-O-${\beta}$-Dglucopyranosyl-($1{\rightarrow}2$)-O-[${\beta}$-D-xylopyranosyl-($1{\rightarrow}3$)]-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-D-galactopyranoside through physicochemical data, spectroscopic methods ($^1H$-NMR, $^{13}C$-NMR, Mass) according references. The quality control of genus Polygonatum were conducted using HPLC quantitative analysis of 14-hydroxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-${\beta}$-D-glucopyranosyl-($1{\beta}4$)-O-${\beta}$-D-galactopyranosyl-26-O-${\beta}$-D-glucopyranoside, 14-hydoxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-[${\beta}$-D-xylopyranosyl-($1{\rightarrow}3$)]-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-D-galactopyranoside in 30 samples collected throughout Korea and China. This method provided a tool for standardization of mix or misusing the commercial Odorati Rhizoma and Polygonati Rhizoma. As a result, contained quantity of 14-hydroxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-D-galactopyranosyl-26-O-${\beta}$-D-glucopyranoside was measured $0.008{\pm}0.006%$ and 14-hydoxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-[${\beta}$-D-xylopyranosyl-(13)]-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-Dgalactopyranoside was measured $0.026{\pm}0.012%$.

• Food Science and Biotechnology
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• v.18 no.1
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• pp.271-274
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• 2009

Cereal $\beta$-glucans, linked essentially by mixed $\beta$-(1,4/1,3) glycosidic bonds, were extracted, purified, and structurally identified. Previously chemical structure of barley $\beta$-glucans was characterized from 3 varieties of 'Gang', 'Ohl', and 'Gwangan', and the (1,4)/(1,3) linkage ratio of the $\beta$-glucans was identical. In this study, $\beta$-glucans from 1 barley ('Chal') and 3 oat ('Ohl', 'Samhan', and 'Donghan') varieties were structurally scrutinized, and the linkage pattern of total 7 cereal $\beta$-glucans was compared. The amount of 2 major 3-O-$\beta$-cellobiosyl-D-glucose (DP3) and 3-O-$\beta$-cellotriosyl-D-glucose (DP4) from barley and oat accounted for only 66.6-73.3 and 68.12-81.89% of water-extractable $\beta$-glucan fractions, and the (1,4)/(1,3) linkage ratios of both barley and oat $\beta$-glucans were within very narrow range of 2.27-2.31 and 2.38-2.39, respectively, among the cultivars tested. Structural difference in the cereal $\beta$-glucans was evident when DP3:DP4 ratio in the $\beta$-glucan structure was compared. As a result, this ratio was significantly greater for barley $\beta$-glucan (2.26-2.74) than for oat (1.54-1.66). Chal-B had the greatest DP3 to DP4 ratio among the samples, which in turn reflected the least amount of (1,4)-linkages.

• Biomolecules & Therapeutics
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• v.23 no.1
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• pp.19-25
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• 2015

Vascular endothelial growth factor (VEGF) is an important regulator of neovascularization. Hypoxia inducible nitric oxide (NO) enhanced the expression of VEGF and thymosin beta-4 ($T{\beta}4$), actin sequestering protein. Here, we investigated whether NO-mediated VEGF expression could be regulated by $T{\beta}4$ expression in HeLa cervical cancer cells. Hypoxia inducible NO production and VEGF expression were reduced by small interference (si) RNA of $T{\beta}4$. Hypoxia response element (HRE)-luciferase activity and VEGF expression were increased by the treatment with N-(${\beta}$-D-Glucopyranosyl)-N2-acetyl-S-nitroso-D, L-penicillaminamide (SNAP-1), to generate NO, which was inhibited by the inhibition of $T{\beta}4$ expression with $T{\beta}4$-siRNA. In hypoxic condition, HRE-luciferase activity and VEGF expression were inhibited by the treatment with $N^G$-monomethyl-L-arginine (L-NMMA), an inhibitor to nitric oxide synthase (NOS), which is accompanied with a decrease in $T{\beta}4$ expression. VEGF expression inhibited by L-NMMA treatment was restored by the transfection with pCMV-$T{\beta}4$ plasmids for $T{\beta}4$ overexpression. Taken together, these results suggest that $T{\beta}4$ could be a regulator for the expression of VEGF via the maintenance of NOS activity.

• Journal of the Korean Chemical Society
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• v.23 no.5
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• pp.307-313
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• 1979

Seven new hydrazine derivatives were prepared from N-Aryllsufonylbenzimidoyl chloride. These were: ${\beta}-({\alpha}-Benzenesulfonamidobenzal)hydrazine\;(II),\;{\beta}-({\alpha}-Benzenesulfonamidobenzal)dimethylmethylenehydrazine\;(III),\;{\beta}-({\alpha}-Benzenesulfonamidobenzal)phenylhydrazine\;(VII),\;{\beta}-({\alpha}-Benzenesulfonamidobenzal)-4-nitrophenylhydrazine\;(VIII),\;{\beta}-({\alpha}-Benzenesulfonamidobenzal)-2,4-dinitrophenylhydrazine\;(IX),\;{\beta}-({\alpha}-Benzenesulfonamidobenzal)dimethylhydrazine\;(X),\;{\beta}-({\alpha}-Benzenesulfonamidobenzal)-p-methylphenylhydrazine\;(XI)$. The structure of these derivatives were identified by elemental analysis, spectral data and other chemical methods. In general, it was found that the yields of these reactions were significantly improved in polar solvent and by the electron attracting substituents in phenylhydrazine.

• Animal cells and systems
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• v.1 no.4
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• pp.589-594
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• 1997

The neuronal voltage-sensitive calcium channels (VSCCs) are multisubunit complexes consisting of $\alpha_1,\;\alpha_2-\delta$ and $\beta$ subunits. Heterologous expression and biochemical studies have shown that the activity of VSCCs is regulated by their $\beta$ subunits in a $\beta$ subunit isoform-specific manner. To elucidate the $\beta$ subunit identity of the P/Q-type calcium channel encoded by an $\alpha_{1A}$ subunit, which is exclusively expressed in the Purkinje and granule cell of the cerebellum, we have examined the spatial and temporal expression patterns of $\beta$ subunits and compared them with those of $\alpha_{1A}$ subunit in the developing rat cerebellum. Reverse transcriptase- polymerase chain reaction (RT-PCR) and Northern blot analysis have shown that $\beta_4$ subunit mRNA was prominently expressed in the cerebellum and much more abundant than any other distinct $\beta$ subunits. RNase protection assay has further demonstrated that the expression of $\alpha_{1A}$ and $\beta_4$ subunits increased during cerebellar development, while the amount of $\beta_2$ and $\beta_3$ mRNAs did not significantly change. In addition, a $\beta_4$ transcript was present in cultured cerebellar granule cells, but not in astrocyte cells, and the level of $\beta_4$ mRNA expression increased gradually in vitro seen as in vivo. Based on the spatial and temporal expression patterns of $\beta_4$ subunit, we conclude that $\beta_4$ may predominantly associate, but probably not exclusively, with the $\alpha_{1A}$ subunit in rat cerebellar granule cells.

• Applied Biological Chemistry
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• v.47 no.4
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• pp.384-389
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• 2004

The purified ${\beta}-mannanase$ hydrolyzed various gums to mannose, ${\beta}-1,4-mannobiose$, $Gal^3Man_4$, and D.P 7 of galactosyl mannooligosaccharide, and isolated from the enzymatic hydrolysate for 24 hrs reaction by activated carbon column chromatography and Sephadex G-25 column chromatography. For the elucidate of antioxidant action of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ and DP 7 of galactosyl mannooligosaccharide and urea derivatives, coloration, reducing power, antioxidant activity and DPPH test were accomplished. The coloration was high at reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and urea. TLC of reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and ureas showed new reaction products, respectively. but except reaction mixture of ${\beta}-1,4-mannobiose$ and urea. The reducing power was high at reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and phenylthiourea. The reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and thiourea showed similar radical scavenging activities on DPPH to activity of AsA. The reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and thiourea, phenythiolurea shown strong antioxidative activites on the oxidation of linoneic acid.

• Applied Biological Chemistry
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• v.39 no.6
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• pp.482-487
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• 1996

Five barley and two oat varieties grown in Korea were investigated for soluble, insoluble, and total $(1{\to}3)$, $(1{\to}4)-{\beta}-D-glucans$. Total and insoluble ${\beta}-glucans$ after extraction of soluble ${\beta}-glucans$ with water were analyzed, and the soluble ${\beta}-glucans$ were calculated as the difference between total and insoluble ${\beta}-glucans$. The total ${\beta}-glucans$ in whole barleys were in a range of $3.3{\sim}5.6%$(average 4.4%), and those in pearled barleys were In a range of $3.3{\sim}7.1%$(average 5.2%). In whole barleys, on average, 54% of the ${\beta}-glucans$ was soluble and in pearled barley 46%. Whole oats contained $3.1{\sim}4.0%$ total ${\beta}-glucans$, and dehulling increased the groat ${\beta}-glucans$ contents to $4.0{\sim}4.8%$. Oats demonstrated considerably higher ${\beta}-glucans$ solubility of 84% than barley. ${\beta}-Glucans$ in barley and oats were rapidly extracted at the beginning of the extraction and almost all of the ${\beta}-glucans$ were extracted after $2{\sim}3 hr extraction. As extraction temperature increased from $23^{\circ}C$ to $45^{\circ}C$, more soluble ${\beta}-glucans$ were extracted. However, solubility of barley ${\beta}-glucans$ decreased at a relatively high temperature of $65^{\circ}C$. Steam-cooking reduced the analytical solubility of barley and oat ${\beta}-glucans$, while roasting seemed to render the ${\beta}-glucans$ of barley more soluble.

• Proceedings of the Korean Biophysical Society Conference
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• 2002.06b
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• pp.32-32
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• 2002

To understand the functional roles of the neuron-specific $\beta$-subunit of large-conductance calcium-activated potassium ($BK_{Ca}$) channel, we isolate the full-length complementary DNA of $\beta$4-subunit from rat brain library and investigated its effects on the function of $\alpha$-subunit (Slo). The deduced amino acid sequence of rat $\beta$4 (r$\beta$4), 210 amino acids in length, was closely related to those of $\beta$4 subunits in other mammalian species but showed only a limited sequence homology to the other $\beta$-subunits, $\beta$1 to $\beta$3.(omitted)d)