• The Korean Journal of Mycology
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• v.25 no.4 s.83
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• pp.330-339
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• 1997

Botrytis cinerea T91-1 has shown to produce at least four different polygalacturonases in a liquid medium containing citrus pectin as a carbon source. One of the enzymes, its molecular weight was estimated as 37 kDa by denatured polyacrylamide gel electrophoresis, was purified by a series of procedures including acetone precipitation, ion exchange, heparin affinity, and reverse phase column chromatographies. By viscometric analysis, the enzyme was revealed as an endo-polygalacturonase. The enzyme activity was inhibited by divalent cations such as $Ca^{2+}$, $Co^{2+}$, and $Cu^{2+}$. Km and Vmax for polygalacturonic acid hydrolysis were 0.33 mg/ml and 28.6 nM/min, respectively. The optimum temperature for enzymatic activity was $55^{\circ}C$ and the enzyme showed optimal pH values between 4.0 and 4.5. The enzyme was stable up to 12 hours in the range of pH 4 to 7 and at the temperature below $30^{\circ}C$. Amino acid sequence from N-terminal up to 6 amino acids determined by Edman degradation showed little homology with polygalacturonases from fungi and plants.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.5
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• pp.659-666
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• 1995

Fucoidans were isolated from Hizikia fusiformis and Sargassum fulvellum and characterized on their chemical properties. Crude fucoidans were extracted at $65^{\circ}C$ for 1hr with acid solution of pH 2.0, and cetylpyridinum chloride was used for partial purification. The yields of partially purified fucoidans were $2.51\%$ for H. fusiformis, and $65\%$ for S. fulvellum, respectively. The partially purified fucoidans were separated into 3 fractions by DEAE-Sephadex A-25 ion exchange column chromatography and the major fractions were refractionated with fractional preripitation with ethanol. $60-70\%$ ethanol precipitated fraction(P-70) of H. fusiformis and $60-65\%$ ethanol precipitated fraction(P-65) of S. fulvellum turned out to be homogeneous by cellulose acetate electrophoresis and gel filtration chromatography. The molar ratios of fucose, galactose, and sulfate In the purified fucoidans were 1 : 0.66 2.74 for H. fusiformis and 1 : 0.24. 1.46 for 5. fulvellum. The averaged molecular weights of the purified fucoidans from H. fusiformis and S. fulvellum were 26,000 and 105,000, respectively.

• Korean journal of applied entomology
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• v.56 no.1
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• pp.19-28
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• 2017

Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.422-427
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• 2006

The culture media from Fomitopsis pinicola were extracted by methanol and examined growth inhibition against Helicobacter pylori. The culture media from 8 days fermentation of F. pinicola showed maximum inhibition activity on H. pylori in 0.25 mg as MIC value. The highest activity against H. pylori by MHCS agar diffusion medium by Fp-P1 in 22.7 mm ID among 3 peaks from methanol fraction of 8 days culture media of Fomitopsis pinicola which purified by ion-exchange chromatography. The Fp-P1 from DEAE-Sephadex A-25 have been analysed by TLC as Fp-T1, Fp-T2 and Fp-T3 by ninhydrin staining. Fp-T3 (Rf value : 0.67) was higher activity against H. pylori in 14.4 mm ID. Fp-T3 was identified as single band by HPLC and TLC. It was assumed to aminosugar by BioLC analysis and TLC staining.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.33-43
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• 2013

A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at $37^{\circ}C$ reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of $40^{\circ}C$ and an optimum pH of 8.0, and the enzyme was stable in the ranges $20-40^{\circ}C$ and pH 6.0-8.0. Enzyme activity was increased by $Ca^{2+}$ and $Co^{2+}$ but inhibited by $Cu^{2+}$, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.

• The Journal of the Korea Contents Association
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• v.19 no.11
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• pp.424-433
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• 2019

The purpose of this study was to evaluate the salivary concentration of Potassium and Magnesium cations and their variation before and after sucrose and glucose rinse, and to investigate the relationship between the levels of each compound. Saliva samples were obtained from 40 subjects before and up to 60 min after intake of a 10% sucrose and glucose solution at 1-month intervals. Potassium and Magnesium in human saliva were determined via anion-exchange chromatography with an anion-suppressed conductivity detector using 12 mM sulfuric acid. The concentrations of Potassium and Magnesium before sucrose rinse were 274.3±77.9 mg/ℓ and 4.5±2.5 mg/ℓ, also, the concentrations of Potassium and Magnesium before glucose rinse were 279.2±62.1 mg/ and 4.8±2.0 mg/ℓ, respectively. Potassium and Magnesium concentrations were significantly decreased (p<0.05) after sucrose rinse. The content of potassium and magnesium in saliva before and after rinsing sucrose and glucose is difficult to standardize or classify, as previous research. The reason for the variation between individuals is large, and easily changed by chemical or physiological stimulation. However, this study was experiment for the purpose of accumulating basic data for saliva.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.320-327
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• 2004

Three hundreds thirty two bacterial colonies which were able to degrade crude oil were isolated from soil sam­ples that were contaminated with oil in Daejeon area. Among them, one bacterial strain was selected for this study based on its higher oil degrading ability, and this selected bacterial strain was identified as Acinetobactor sp. B2 through physiological-biochemical tests and analysis of its 16S rRNA sequence. Acinetobactor sp. B2 was able to utilize various carbohydrates but did not utilize trehalose and mannitol as a sole carbon source. Acinetobactor sp. B2 showed a weak resistance to antibiotics such as kanamycin, streptomycin, tetracycline and spectinomycin, but showed a high resistance up to mg/ml unit to heavy metals such as Ba, Li, Mn, AI, Cr and Pb. The optimal growth temperature of Acinetobactor sp. B2 was $30^{\circ}C.$ The lipase produced by Acinetobactor sp. B2 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 60 kDa and condition for the optimal activity was observed at $40^{\circ}C$ and pH 10, respectively. The activation energy of lipase for the hydrolysis of p­nitrophenyl palmitate was 2.7 kcal/mol in the temperature range of 4 to $37^{\circ}C,$ and the enzyme was unstable at the temperature higher than $60^{\circ}C.$ The Michaelis constant $(K_m)\;and\;V_{max}$ for p-nitrophenyl palmitate were 21.8 uM and $270.3\;{\mu}M\;min^{-1}mg^{-1},$ respectively. This enzyme was strongly inhibited by 10 mM $Cd^{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},$ EDTA and 2-Mercaptoethalol.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.269-274
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• 2004

Serratia proteamaculans isolated from the midgut of a spider formed big halos around the bacterial colonies, indicating that the bacterial strain produces an extracellular protease. Activity staining of the extracellular pro­tein fractions using zymogram also demonstrated that the major protein with an estimated molecular mass of 52 kDa contained a high proteolytic activity. The protease was purified to near electrophoretic homogeneity from the culture supernatant after filtration and ion exchange and size exclusion chromatography. The purified enzyme had a relatively high proteolytic activity between pH 6.0 and 10.0 and at broad temperature range. The proteolytic activity of the enzyme was not inhibited by phenylmethylsulfonyl fluoride but strongly inhibited by 1, 10-phenanthroline and EDTA. The activity also was dependent on the presence of $Ca^{++}\;and\;Zn^{++}$ ions. These observations indicate that the enzyme is a metalloprotease.

• Korean Journal of Food Science and Technology
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• v.34 no.5
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• pp.879-884
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• 2002

Cold and hot water fractions of Diospyros kaki were screened to determine its anti-complementary activity. Flour of Diospyros kaki leaf (250 g) was boiled at $100^{\circ}C$ for 3 h and passed through a membrane of 10 kDa molecular weight (DK-0). DK-0 was precipitated with ethanol and refluxed with methanol to obtain the crude polysaccharide (DKC). DKC-1 was isolated by ion exchange chromatography on DEAE-Toyopearl 650C, and DKC-1c was purified from DKC-1 by size exclusion chromatography on Bio gel P-60. The anti-complementary activities of DKC-1c at $1000\;{\mu}g/mL$ were 85.4 and 61.1% via whole and alternative pathways, respectively. DKC-1c was determined as a neutral polysaccharide composed of glucose (29.0 mol.%), arabinose (24.3 mol.%), and galactose (16.2 mol.%) with the molecular weight of 66.6 kDa. Results of agarose gel immunoelectrophoresis revealed DKC-1c, as a complement activator, cleaved C3 into C3a and C3b via both pathways.

• Journal of the Korean Society of Food Science and Nutrition
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• v.19 no.6
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• pp.625-635
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• 1990

The purpose of this study was focused on investigation of biochemical properties of amylases in germinating corn(Zea mays L.) the amylase(I), (II) and (III) from germinating corn seeds were partially purified by ammonium sulfate precipitation, DEAE-Sephadex A-50 ion exchange column chromatography and Sephadex G-100 gel filtration. The last step was effective for separation of the corn amylases to a homogeneous slate. the purified amylase(I) was identified as a kind of $\alpha$-amylase from the fact that 5％ starch solution was hydrolysed into mainly maltose and maltotetrose by it, and amylase(II) and amylase(III) were enzymes producing maltotetrose as main product. The molecular weight and specific activity of the amylase(I), (II) and (III) were determined to be 54,000 and 70.47 unit/mg, 39,000 and 62.98 unit/mg, and 51,000 and 80.39 unit/mg, respectively. It showed a tendency to increase the amylases activities in presence of Ba, Ca, Co and Fe groups, but inhibits in that of Ag, Sn, Hg and Zn groups, and amylase(I), (II) and (III) remained stable at pH 5-6 and 2$0^{\circ}C$ for 40 days in containing of 1 mM CaCl$_2$. The optimum pH and optimum temperatures were pH 6, pH 5 and pH 6 and 35$^{\circ}C$, 55$^{\circ}C$ and 55$^{\circ}C$, respectively. These results suggest that the amylase(I), (II) and (III) were different amylases.