• Journal of Life Science
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• v.22 no.2
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• pp.142-147
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• 2012

The sporulation-specific glucoamylase (SGA) of Saccharomyces diastaticus is known to be produced in the cytoplasm during sporulation. For the purpose of proving that SGA has secretory potential, we constructed a hybrid plasmid, pYESC25, containing the promoter and the putative signal sequence of the SGA fused in frame to the endo-1,4-${\beta}$-D-glucanase (CMCase) gene of Bacillus subtilis without its own signal sequence. The recipient yeast strain of S. diastaticus YIY345 was transformed with the hybrid plasmid. CMCase secretion from S. diastaticus harboring pYESC25 into culture medium was confirmed by the formation of yellowish halos around transformants after staining with Congo red on a CMC agar plate. The transformant culture was fractionated to the extracellular, periplasmic, and intracellular fraction, followed by the measurement of CMCase activity. About 63% and 13% enzyme activity were detected in the culture supernatant (extracellular fraction) and periplasmic fraction, respectively. Furthermore, ConA-Sepharose chromatography, native gel electrophoresis, and activity staining revealed that CMCase produced in yeast was glycosylated and its molecular weight was larger than that of the unglycosylated form from B. subtilis. Taking these findings together, SGA has the potential of secretion to culture medium, and the putative signal sequence of SGA can efficiently direct bacterial CMCase to the yeast secretion pathway.

• Korean Journal of Veterinary Research
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• v.32 no.2
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• pp.181-194
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• 1992

Two hundred and eighty nine strains of Yersinia enterocolitica isolated from healthy pigs were tested for the presence of 40~50 Megadalton virulence-associated plasmids and plasmidmediated in vitro virulence-associated properties, i.e., congo red uptake, calcium dependency, autoagglutination, CRMOX reaction, crystal violet binding and pyrazinamidase reaction. The correlationships between in vitro virulence-associated properties and the presence of 220 Kdalton outer membrane protein(OMP) were examined in strains with or without virulence-associated plasmids. The correlationships between the presence of plasmids on the production of the OMP and the expression of in vitro virulence-associated properties were studied with $CRMOX^+$ strains and acridine orangecured $CRMOX^-$ mutants. The results were as follows : 1. Of the in vitro virulence-associated tests with 289 strains of Y enterocolitica, 275 strains (95.2%) were positive for pyrazinamidase test, and followed by in order of crystal violet binding test, 226 (79.2% ) ; CRMOX test, 190 (65.7%) ; autoagglutination test, 1.85(64.0%) : calcium dependency test, 86 (29.8%) and congo red uptake test, 47(16.3%). 2. The correlationship between autoagglutination and CRMOX test(r=0.90) was highly significant (p<0.01). 3. In 190 strains(65.7%) bearing the virulence-associated plasmids(MW 40~50 Mdalton), the correlation between the presence of plasmids and their in vitro virulence-associated properties were highest with CRMOX test(r=0.93) and followed by in orders of AAG test(0.81), CV test(0.46), PYZ test(0.37) and CD test(0.18), but no correlationship between the presence of plasmids and CR test(-0.11). 4. The $CRMOX^+$ strains produced the 220 Kdalton OMP when they were cultured at $37^{\circ}C$, but not at $26^{\circ}C$. The presence of 220 Kdalton OMP was correlated significantly with in vitro virulence properties and the presence of virulence-associated plasmid, respectively. 5. In the isogenic $CRMOX^-$ mutant strains, of which plasmid were cured by treatment with acridine orange not only in vitro virulence-associated properties(CR 100%, CD 100%, AAG 82.6%, CV 58.3%) disappeared but also 220 Kdalton OMP(100%) was not produced. These results indicate that the positive CRMOX reaction is plasmid-mediated and the CRMOX test is potential as an in vitro virulence tests with Y enterocolitica.

• Korean Journal of Microbiology
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• v.35 no.4
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• pp.277-282
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• 1999

A bacterium producing the extracellular cellulase was isolated from cattle feces and screened as cellulase activity was excellent upon congo red straining method and activity measurements. Isolate was identified as Bacillus subtilis CH-10 on the basis of morphological and biochemical properties as well as cellular fatty acids composition. The enzyme which the isolate secretes had the optimum initial pH and temperature for its induction was 7.5 and 50${\circ}C$, respectively. The maximum CMCase activity in crude enzyme solution was observed at pH 7.5 and 75${\circ}C$ and was stable for pH 7.5 to 9.0 to maintain 70% activity. When the isolate was cultured in CMC media at 37${\circ}C$ for 24 hrs, CMCase and FPase activity was 1.13 U/㎖and 0.16U/㎖, respectively whereas Avicelase and ${\beta}$-glucosidase activity was not detected. When crude supernatant was used for zymogram, three major bands, cel 1, cel 2 and cel 3, were detected approximately 39, 41 and 57 KDa, respectively on CMC-SDS-PAGE.

• Applied Biological Chemistry
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• v.44 no.2
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• pp.109-115
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• 2001

The foliar uptake of dimethomoiph induced by several nonionic surfactants was measured in order to study the correlations between the uptake rate of dimethomorph and the fungicidal activity to cucumber downy mildew. Dimethomorph was not absorbed in cucumber leaf in the absence of activator surfactant. And the curative effect of dimethomoiph WP to cucumber downy mildew was very low under the concentration of 250 ${\mu}g/ml$. But dimethomorph uptake was remarkably enhanced by addition of nonionic surfactants, such as polyoxyethylene cetyl ether, polyoxyethylene oleyl ether, and polyoxyethylene stearyl ether. And the curative effect to cucumber downy mildew was enhanced with proportion to uptake rate of dimethomorph. The protective effect to cucumber downy mildew, however, tends to decrease with the increase of foliar uptake of dimethomorph. The uptake rate of dimethomorph to cucumber leaf was proportional to the content of polyoxyethylene cetyl ether in formulation, but was decreased with dilution.

• The Korean Journal of Pesticide Science
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• v.14 no.3
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• pp.272-279
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• 2010

KNF-1002, a new fungicide candidate, is very effective for protecting crops against plant diseases, but its curative activity against barley powdery mildew is negligible due to its weak penetrability into plant leaf. To select the most efficient activator and, at the same time, spreade-sticker, foliar uptake and deposit of aqueous spray formulations containing non-ionic or anionic surfactants and fatty acid alkyl esters as an adjuvant were assessed by using Congo Red method. In the absence of activator, only 0.1% of the applied active ingredient was absorbed by barley leaves 24 h after spraying with an aqueous acetone containing KNF-1002 100 mg/L. But, non-ionic surfactants (500 mg/L), such as heptaethylene glycol monooctadecenyl ether (OE-7), dodecaethylene glycol monohexadecyl ether (CE-12), so facilitated KNF-1002 uptake that the uptake was increased up to 48.5%. To wheat plant, the addition of surfactants in spray solution of KNF-1002 also increased the foliar uptake and deposition of active ingredient, but its efficiency varied according to the kind of fatty alcohol moiety of polyoxyethylene surfactant. KNF-1002 formulations containing nonaethylene glycol monododecyl ether (LE-9) as an activator and spreader-sticker showed remarkable increases of fungicidal activity against barley powdery mildew.

• Korean Journal of Microbiology
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• v.42 no.1
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• pp.34-39
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• 2006

Wood-rot fungi produce extracellular lignin-degrading enzymes, the best known of which are lignin peroxidase, Mn-peroxidase and laccase. In this experiment, some of them produced all of three enzymes. Many other wood-rot fungi produced one or two of those enzymes with various combinations. In this experiment, we tried to clarify the relationship between the pattern of enzyme production and degradative activity of several dye compounds. From the 36 strains of 23 species of wood-rot fungi, Mn-peroxidase activity was found in 30 strains of the fungi tested, whereas the activity of lignin peroxidase and laccase was detected in 11 strains and 12 strains of species, repectively, in Kirks low nitrogen media. In relation to the activity of lignin degrading enzymes and degradation of dye compounds, the white-rot fungi with three kinds of enzymes tested showed the best dye decolorizers. The fungi with Mn-peroxidase activity only decolorized poly R-478 and remazol brilliant blue R dye in proportion to the enzyme activity, while methylene blue, bromophenol blue and congo red dye were degraded in regardless of enzyme activity. Those dyes were degraded in relation to the growth rate of mycelium. Brown-rot fungi did not degrade all the dye compounds except bromophenol blue, in spite of moderate growth rate.

• Korean Journal of Microbiology
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• v.45 no.1
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• pp.63-68
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• 2009

The exo-polysaccharide producing microorganism, Aureobasidium pullulans IMS-822, was isolated and identified from soil. The viscosity-average molecular weight of exo-polysaccharide was calculated as $8.9{\times}10^5$ by Mark-Houwink equation. The sugar component of exo-polysaccharide was determined as glucose by HPLC analysis. The IR spectra indicated that the exo-polysaccharide has an absorption peak at 890 $cm^{-1}$ for the ${\beta}-configuration$ of D-glucan. The $^{13}C$ NMR signal at ${\delta}$ 86.62 ppm arose from the substituted C-3 of glucose. The signal at ${\delta}$ 72.11 ppm was assigned to C-6 of branched ${\beta}-(1{\to}3)-D-glucosyl$ residues. Viscosity and Congo red reaction indicated that {\beta}-(1{\to}3)(1{\to}6)-glucan$ produced by A. pullulans IMS-822 has a highly ordered hydrogen-bond dependent conformation in aqueous solution, which collapses in strong alkaline solution.

• The Korean Journal of Mycology
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• v.26 no.1 s.84
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• pp.8-15
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• 1998

The present research was undertaken to investigate the activity of ligninolytic enzymes and the decolorization capability of some dyes with Irpex zonatus BN2, isolated from nature and identified. For the assay of enzyme activities, the isolate did not produce lignin peroxidase (LiP) and veratryl alcohol oxidase (VAO), but laccase and manganese dependent peroxidase (MnP). While the activity for MnP was low $(61.6\;nmol/mg{\cdot}protein)$, its laccase activity was very high $(1185.9\;nmol/mg{\cdot}protein)$. Moreover, laccase had appeared earlier than MnP. When the isolate was incubated with each dye for 10 days, the decolorization rates of azo dyes, such as orange II, orange G, tropaeolin O and congo red were 98.0%, 97.4%, 99.0% and 95.3%, respectively. In case of heterocyclic dyes, eosin Y, toludine blue, methyl blue and azur B were 97.4 %, 98.7%, 99.9% and 94.0% respectively. Finally the results of triphenylmethane dye such as basic fuchsin, malachite green and crystal violet were 98.5%, 95.7% and 99.4%, respectively. The results suggest that laccase of Irpex zonatus BN2 should be played an important role in the decolorization of the dyes.

• Applied Biological Chemistry
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• v.22 no.3
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• pp.166-172
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• 1979

A series of experiments was planned for practical application of rhizobia in grass lands in Korea. This is the report for the studies mainly on the isolation and characterization of alfalfa nodule bacteria, and on the assessment of their nodulation abilities and nitrogen fixation capacities. 1. Total number of 47 strains was isolated from nodules which were taken from alfalfa grown in Daekwanyong, Cheju and other places. 2. Morphological and cultural characteristics of the strains were studied, and attempts. were also made to investigate their antigenic properties and to demonstrate lysogenic strains. The results were; i) the isolates varied in their cultural characteristics on yeast mannitol broth and agar, and in degree of congo red absorption; ii) similarities in their antigenic prorerties were found between the strains: SU 47/M-11, M-13/M-15, and M-3/M-5; iii) no lysogeny was found in the strains. 3. Plant infection test by test tube method in light room were carried out to elucidate the ability of the strains to nodulate Luna alfalfa and of the capacity of such nodules to fix atmospheric nitrogen. The isolates were grouped info non-invasive, ineffective, or effective to the legumes. Those strains which produced effective nodules, supporting similar/higher level of growth as nitrate control were: M-8, 9, 14, 20, 21, 25 and 34.

• Membrane Journal
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• v.31 no.3
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• pp.228-240
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• 2021

The organic solvent robust polybenzimidazole (PBI) membranes were prepared as organic solvent nanofiltration (OSN) membrane for the recycling of alcoholic solvents using non-solvent induced phase separation with different dope solution concentration and coagulant composition of water/ethanol mixtures to control the membrane morphology and permeation performance. Investigation on crosslinking of polybenzimidazole indicated that the membrane crosslinked with dibromoxylene (DBX) had enough mechanical strength and solvent resistance to be applied as a OSN membranes. The crosslinked PBI membrane prepared by more than 20wt% dope concentration coagulated in water showed a rejection of > 90% to Congo Red (MW of 696.66 g/mol) while pure ethanol permeances was more than 22.5 LMH/bar at 5 bar. Investigation on coagulant composition indicated that ethanol permeance through crosslinked PBI OSN membrane increased with increasing of ethanol concentration in water/ethanol mixture coagulants.