• Applied Biological Chemistry
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• v.35 no.3
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• pp.165-169
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• 1992

Two types of $endo-{\beta}-1,3-glucanases$ were purified from green malt and their basic characteristics were studied. Molecular weights of glucanase I and glucanase II were estimated, by electrophoresis, to be 35,000 and 28,000, respectively. Purified glucanase I and II showed the highest activity at pH $5.0{\sim}7.0$ and $5.0{\sim}8.0$, respectively. The optimal temperature of purified glucanase I and II was $40^{\circ}C$. Purified glucanase I and glucanase II were stable at $40^{\circ}$ for 60 min and at $50^{\circ}$ for 30 min. All enzymes were inactivited by $AgNO_3$ and $HgCl_2$ while those were not activated by various compounds tried. Km values of glucanase I and II were 1.03 mg/ml, 1.20 mg/ml, respectively.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.2
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• pp.250-254
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• 1995

Isolated barley aleurone layers were used to examine response of (1-3)- and $(1-3,1-4)-{\beta}-glucanases{\;}to{\;}GA_3$. Protein content and levels of (1-3)- and $(1-3,1-4)-{\beta}-glucanases$ increased in the presense of added $GA_3$. However, (1-3)- and $(1-3,1-4)-{\beta}-glucanases$ showed different response to $GA_3$ in their production and secretion patterns. $(1-3,1-4)-{\beta}-glucanases$ showed higher increase in enzyme activity than $(1-3)-{\beta}-glucanase$ in the early stage of$GA_3$treatment. Secretion of enzyme by $GA_3$ into the surrounding medium was more enhanced in $(1-3,1-4)-{\beta}-glucanases$ than in $(1-3)-{\beta}-glucanase$, The differential response of the enzymes might be related to the physiological role of the enzymes in germination of barley grain.

• Journal of Life Science
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• v.24 no.10
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• pp.1046-1054
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• 2014

Beta-1,3-glucanase is widely used in various biotechnological and industrial processes, with over-production required to enable versatile utilization. We examined the overexpression of ${\beta}$-1,3-glucanase (EXGA) from Aspergillus oryzae using ${\delta}$-sequence-mediated integration. We constructed $pRS{\delta}$-exgA and $pRS{\delta}K$-exgA plasmids for integration of the EXGA gene into various chromosomes of Saccharomyces cerevisiae. These plasmids contain the ADH1 promoter for constitutive expression, a signal sequence (exoinulinase signal sequence [INU1 s.s]) for secretory production, and a ${\delta}$-sequence for integration of ${\beta}$-1,3-glucanase. The $pRS{\delta}$-exgA plasmid was transformed into the S. cerevisiae $BY4742{\Delta}exg1$ strain, and ${\beta}$-1.3-glucanase was stably overexpressed and secreted. Another plasmid, $pRS{\delta}K$-exgA, was introduced into the S. cerevisiae $BY4742{\Delta}exg1$ (YKY082) strain, and overexpression of ${\beta}$-1,3-glucanase was examined by inducible integration under geneticin selection. The activity of ${\beta}$-1,3-glucanase increased in accordance with a rise in the geneticin concentration, with 0.8 mg/ml of geneticin suitable for overexpression of ${\beta}$-1,3-glucanase. Subsequently, $pRS{\delta}K$-exgA was repeatedly transformed for sequential ${\delta}$-integration. The activity of ${\beta}$-1,3-glucanase reached about 0.063 unit/ml/$OD_{600}$, 0.095 unit/ml/$OD_{600}$, 0.131 unit/ml/$OD_{600}$ and 0.165 unit/ml/$OD_{600}$ by the first, second, third, and fourth round of integration, respectively. According to the increase in the activity of ${\beta}$-1,3-glucanase by sequential ${\delta}$-integration, the copy number (integration rate) of the EXGA gene also increased in various chromosomes. These results suggest that recombinant ${\beta}$-1,3-glucanase activity can be sequentially increased by repeated ${\delta}$-sequence integration.

• KSBB Journal
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• v.26 no.5
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• pp.427-432
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• 2011

In this study, a EXGA gene code for exo-β-1,3-glucanase from Aspergillus oryzae was overexpressed and secretory produced in Saccharomyces cerevisiae. To overexpress the β-1,3-glucanase, pGInu-exgA and pAInu-exgA plasmids having GAL10 and ADH1 promoter, respectively, and exoinulinase signal sequence (Inu s.s) were constructed and introduced in S. cerevisiae SEY2102 and 2805. The recombinant β-1,3-glucanase was successfully expressed and secreted into the medium and the β--1,3-glucanase activity in 2102/pGInu-exgA and 2102/pAInu-exgA strain were 5.01 unit/mL and 4.09 unit/mL, respectively. In the 2805/pGInu-exgA and 2805/pAInu-exgA strain, the β-1,3-glucanase activity showed 3.23 unit/mL and 3.22 unit/mL, respectively. Secretory efficiency in each strain reached 95% to 98%. Subsequently, the recombinant β1,3-glucanase was used for ethanol production. Ethanol productivity in 2102/pAInu-exgA strain was 0.83 g/L when pre-treated Laminaria japonica which has initial reducing sugar of 1.4 g/L was used as substrate. It is assumed that the polysaccharides of Laminaria japonica was effectively saccharified by recombinant β-1,3-glucanase, resulting in increase of ethanol productivity. These results suggested that recombinant β-1,3-glucanase was efficiently overexpressed and secreted in S. cerevisiae SEY2102 as host strain by using ADH1 promoter-Inu s.s system.

• Microbiology and Biotechnology Letters
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• v.21 no.3
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• pp.263-268
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• 1993

Seventeen strains were isolated from soil, cattle rumen, cereal sewage dregs, insect on agar plate containing insoluble glucan as a sole carbon source from immobilized Streptococcus mutans, which produced alpha-1,3 glucanase for lysis of dental plaque. Among these strains isolated from soil, SW-522 and SW-713 that had appeared to produce the high level of alpha-1,3 glucanase, degraded insoluble glucan from S. mutans 97.6% and 49.4%, respectively in 5 hours. The activity of crude alpha-1,3 glucanase from SW-522 was 1.3mg insoluble glucan/min.mg protein. This enzyme was entirely degraded insoluble glucan on glass tube which produced by S. mutans in TH medium with 5% sucrose.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.2
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• pp.121-127
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• 1994

A procedure for the direct detection of (1-3)-$\beta$-glucanase isozymes in electrophoresis gels was developed. The procedure employed the commercial preparation of AZCL-pachyman as a chromogenic substrate for (1-3)-$\beta$-glucanases. The procedure detected the three basic isozymes which have been known to be expressed in germinating barley kernels. A major acidic and a minor isozymes were also detected in germinating kernels. The procedure was proved to be fast, simple and sensitive enough to be used for the analysis of the expression of (1-3)-$\beta$-glucanase isozymes in plant tissues. The detection limit of the procedure for the commercial preparation of Penicillium (1-3)-$\beta$-glucanase was estimated to be as low as 50$\mu$U. The procedure could be used for the investigation of (1-3)-$\beta$-glucanases in laboratories facilitated with ordinary equipments and research personnel.

• Proceedings of the Korean Society of Crop Science Conference
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• 1992.05a
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• pp.22-23
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• 1992

• Proceedings of the Korean Society of Crop Science Conference
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• 1992.05a
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• pp.24-25
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• 1992

• Proceedings of the Korean Society of Crop Science Conference
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• 1992.05a
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• pp.20-21
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• 1992

• Journal of Life Science
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• v.21 no.1
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• pp.68-73
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• 2011

$\beta$-1,3-glucanase from Pyrococcus furiosus was applied for the saccharification of laminarin, which is a major oligo-saccharide component of brown algae, and the reaction mixture produced from laminarin was utilized as a substrate for alcohol fermentation using yeast. To prepare the recombinant $\beta$-1,3-glucanase, a $\beta$-1,3-glucanase gene was overexpressed in Escherichia coli and purified. Laminarin was degraded to an oligo- and mono-saccharide, such as glucose, after reaction with the purified recombinant $\beta$-1,3-glucanase, and the products after enzymatic treatment were confirmed by TLC and HPLC analysis. Decomposed laminarin after enzyme reaction was only added to the medium as a C-source for yeast alcohol production reaction. 0.3% alcohol production was detected from the cultured broth by gas chromatography after 48 hr of incubation. Further evaluation for optimal conditions of saccharification and alcohol fermentation can be suggested, as well as the possibility of using this enzymatic method to produce ethanol using laminarin.