• Applied Chemistry for Engineering
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• v.23 no.2
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• pp.164-168
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• 2012

We investigated for the production of biological bio-ethanol from Laminaria japonica using the hydrolysis reaction of enzymes and organic acids and the polysaccharide content was also analyzed. The composition of the polysaccharide was characterized as 65.99% alginate, 6.24% laminaran and 27.77% mannitol. The optimum concentration for reducing the sugar conversion by Laminaria japonica was found to be 1.874 g/L at an acetic acid concentration of 1.5%, $121^{\circ}C$ for 60 min, and for an ascorbic acid of 2.0%, 4.291 g/L was produced in the same condition. The enzyme hydrolysis such as alginate lyase and laminarinase contained the maximum 2.219 g/L reducing sugar. In the result of ethanol fermentation using hydrolysate of Laminaria japonica, the organic acid treatment showed a high of reducing sugar yield, but decreased the ethanol yield, and then the maximum ethanol production obtained was 1.26 g/L using the mixed treated of enzyme.

• Microbiology and Biotechnology Letters
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• v.14 no.6
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• pp.467-471
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• 1986

Yeast cell wall lytic enzyme was purified from Fusarium moniliforme by ammonium sulfate fractionation and gel column chromatography. The lytic activity was found to consist of three enzyme activities which were resolved on Sephadex G-100. The first peak on chromatogram exhibited proteolytic, lytic and laminarinase activities, and the second had both lytic and laminarinase activities, whereas the third peak was shown to contain lytic activity only. Three enzyme activities showed the synergistic effect and reducing agents accelerated the yeast roil wall lysis. This indicates that lytic, proteolytic and laminarinase activity acted cooperatively in the lysis of intact cells. Tannic acid precipitate of crude enzyme constituted of three enzyme activities had a high lytic activity on viable yeast cell and has proved useful in yeast protoplast formation.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.81-88
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• 1990

For the selection of powerful antagonistic bacterium for biological control of soilborne Fusarium solani causing root rot of many important crops, the best YPL-1 strain was selected among 300 strains of bacteria isolated from rhizosphere in ginseng root rot-suppressive soil. The strain was identified to be a species to Pseudomonas stutzeri. With in vitro fungal inhibition tests, antagonistic substance of P. stutzeri YPL-1 against F. solani was presumed to be heat unstable, macromolecular substances such as protein. Also, it was shown that antifungal activity of P. stutzeri YPL-1 increased in proportion to its chitinase production. P. stutzeri YPL-M122 (chi-, lam -) which was deprived of the productivity of chitinase and laminarinase by NTG mutagenesis had lost antifungal activity, completely. And P. stutzeri YPL-MI53 (chi-) had only 4.1% of its antifungal activity. P. stutzeri YPL-1 was not able to produce any extracellular siderophore in iron-deficent minimal medium. It is confident that the antifungal mechanism of P. stutzeri YPL-1 for biocontrol of F. solani depends on lysis rather than antibiosis :the mechanism of lysis appears to involve enzymatic degradation of the cell will components of F. solani by hydrolytic enzymes of more chitinase and less laminarinase.

• Korean Journal of Environmental Biology
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• v.22 no.1
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• pp.184-191
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• 2004

To investigate bacteria with algalytic activities against Anabaena cylindrica when water blooming occurs and to study enzyme profiles associated with alga-lytic activity, various bacterial strains were isolated from surface waters and sediments in eutrophic lakes or reservoirs in Korea. Among 178 isolates, only nine isolates exhibited lytic abilities against A cylindrica on the agar plates, and then the isolate AK-13 was selected as the strongest in lysing the cyanobacterium A. cytindrica. The strain AK-13 was characterized and identified as Sinorhizobium sp. based on fatty acid methyl ether profiles and 16S rDNA sequence. According to the results of the enzyme assays, in the strain An-13 of Sinorhizobium sp., alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase was produced, namely CMCase, laminarinase and protease were highly active. None of glycosidase was produced. Therefore, enzyme systems of Sinorhizobium sp. AK-13 were very complex to degrade cell walls of A. cylindrica. The peptidoglycans of A. cylindrica mat be hydrolyzed and metabolized to a range of easily utilizable monosaccharides or other low molecular weight organic substances by Sinorhizobium sp. AK-13.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.5
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• pp.382-390
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• 2006

Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

• 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.

• The Korean Journal of Malacology
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• v.27 no.1
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• pp.9-14
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• 2011

This study was examined digestive enzyme activity in the crystalline style of the three species of bivalves. Bivalves used in this study were Tegillarca granosa (n=61), Mytilus galloprovincialis (n=30) and Saxidomus purpuratus (n=30) and collected from southern coast of Korea on May 2010. Digestive enzymes activities in the crystalline style were assayed in spectrophotometer. Amylase and cellulase occupied approximately 90% of digestive enzyme in crystalline style of T. granosa, M. galloprovincialis, and S. purpuratus. And protease activity in crystalline style of T. granosa, M. galloprovincialis and S. purpuratus showed the lowest values to 0.02, 0 and 0.08%, respectively. Digestive enzyme activity in crystalline style of three species was measured in the order of cellulase > amylase > chitinase > laminarinase.

• 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.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.299-301
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• 2018

Microbulbifer agarilyticus GP101 was isolated from the gut of a marine invertebrate Turbo cornutus and capable of degrading polysaccharide such as agar, alginate, and ${\kappa}$-carrageenan constituting algal cell wall. To obtain genomic basis of polysaccharide-degrading activity, we sequenced genome of strain GP101. The genome consists of 4,255,625 bp, 3,458 coding sequences with 55.4% G + C contents. BLASTP search revealed the presence of seven agarases, five alginate lyases, ten glucanases, four chitinases, two xylanases, one ${\kappa}$-carrageenase, and one laminarinase. The genomic data of strain GP101 will provide potential uses in the bioconversion process of diverse polysaccharide into bioenergy and biochemicals.

• Applied Biological Chemistry
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• v.34 no.1
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• pp.54-60
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• 1991

Serratia marcescens CK-3, decomposing chitin which is a mar component of cell wall in phyitopathogenic fungi, was isolated from the continuous cropping rhizosphere of pepper and cucumber and its enzymatic property was examined. S. marcescens CK-3 was found tn have an tagonistic effects against, Fusarium axysporum and Rhizoctonia solani and to have complex enzyme system such as chitinase, laminarinase, and proteinase. The preferable composition of the medium for production of chitinase was fond and was as follows : colloidal chitin 1.5%, tryptone 0.5%, glucose 1.0%, peptone 0.2%, $MgSO_4{\cdot}7H_2O\;0.1%,\;K_2HPO_4\;0.1%,\;and\;NaCl\;0.1%$(w/v), pH 6.8. The maximum enzyme production was observed after culture of 72 hours at $30^{\circ}C$ using a medium containing the above chemical composition. The optimal pH and temperature for in vitro activity of chitinase from S. marcescens CK-3 were pH 7.5 and $50^{\circ}C$, respectively. The enzyme activity in-creased by metal ions such as$Ag^+$ and $Mn^{++}$.