• Journal of Microbiology
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• v.41 no.4
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• pp.271-276
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• 2003

The inoculation of some microorganisms into a microcosm containing soil from a barren lakeside area at Lake Paro in Kangwon-do enhanced plant growth significantly. The direct and viable counts of soil bacteria and soil microbial activities measured by electron transport system assay and fluorescein diacetate hydrolysis assay were higher in inoculated soil. The plant growth promoting effect of this inoculation may be caused by phytohormone production and the solubilization of insoluble phosphates by the inoculated bacteria. Three inoculated strains of Pseudomonas fluorescens produced several plant growth promoting phytohormones, including indole-3-acetic acid (auxin), which was confirmed by thin layer chromatography and GC/MS. P. fluorescens strain B16 and M45 produced 502.4 and 206.1 mg/l of soluble phosphate from Ca3(PO4)2 and hydroxyapatite, respectively. Bacillus megaterium showed similar solubilization rates of insoluble phosphates to those of Pseudomonas spp. We believe that this plant growth promoting capability may be used for the rapid revegetation of barren or disturbed land.

• Bulletin of the Korean Chemical Society
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• v.1 no.1
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• pp.10-17
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• 1980

Penicillin amidohydrolase was partially purified from the fermented broth of Bacillus megaterium, and was immobilized on nylon fiber. The surface area of nylon fiber was increased by roughening it with fine sand and activated by acid treatment. The free amino groups on the nylon fiber exposed by such treatment were then utilized to immobilize the penicillin amidase. Enzymatic properties of penicillin amidohydrolase immobilized on the nylon fiber by covalent bonding and cross linking with glutaraldehyde were studied and compared with those of soluble enzyme. The optimal pH and temperature profile of immobilized enzyme showed only slightly broader peaks, and the values of kinetic constants, $K_m$, $K_{ia}$, and $K_{ip}$, of the immobilized enzyme are only slightly greater than those of the soluble enzyme. These results suggest that the mass transfer effect on the reaction rate for the penicillin amidase immobilized on nylon fiber is not so significant as the enzyme immobilized on some other support material like bentonite. The experimental results of batch reaction agreed well with the results of computer simulation for both the immobilized and soluble enzyme systems, confirming the validity of the rate equation derived which was based on the combined double inhibition by two reaction products.

• The Plant Pathology Journal
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• v.18 no.1
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• pp.36-42
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• 2002

Resistance of perilla varieties to Botrytis cinerea LVF12 was evaluated, while antagonistic bacteria were selected and tested for their efficacy towards biological control of gray mold rot caused by B. cinerea. Among 11 perilla varieties tested for disease resistance, Milyang variety showed some degree of resistance, while the rest of varieties showed no resistance. Among 250 bacterial isolates collected from perilla loaves and rhizosphere of perilla plants, six isolates showed high levels of inhibitory effect on mycelial growth and conidial germination of B. cinerea in in vitro test. Using the pot test in growth chambers these isolates showed high levels of disease suppression, with Nl isolate showing 95.3% of control value and N4 isolate showing 90.8% of control value. Further test was performed to evaluate the two isolates ability for disease prevention and/or disease therapy, and results showed almost 100% of control vague. Isolates Nl and N4 were identified as Bacillus licheniformis and 5. megatepium, respectively, according to Bergey's manual, API 20E and 50CHB test kit, and Transmission electron microscope.

• The Plant Pathology Journal
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• v.39 no.1
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• pp.88-107
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• 2023

In the present investigation, bacterial isolates from infected apple trees causing apple canker during winter were studied in the northern Gyeongbuk Province, Korea. The pathogen was identified as Pseudomonas syringae pv. syringae (Pss) through various physiological and biochemical characterization assays such as BIOLOG, gas chromatography of fatty acid methyl esters, and 16S rRNA. Bioassays for the production of phytotoxins were positive for syringopeptin and syringomycin against Bacillus megaterium and Geotrichum candidum, respectively. The polymerase chain reaction (PCR) method enabled the detection of toxin-producing genes, syrB1, and sypB in Pss. The differentiation of strains was performed using LOPAT and GATTa tests. Pss further exhibited ice nucleation activity (INA) at a temperature of -0.7℃, indicating an INA⁺ bacterium. The ice-nucleating temperature was -4.7℃ for a non-treated control (sterilized distilled water), whereas it was -9.6℃ for an INA^- bacterium Escherichia coli TOP10. These methods detected pathogenic strains from apple orchards. Pss might exist in an apple tree during ice injury, and it secretes a toxin that makes leaves yellow and cause canker symptoms. Until now, Korea has not developed antibiotics targeting Pss. Therefore, it is necessary to develop effective disease control to combat Pss in apple orchards. Pathogenicity test on apple leaves and stems showed canker symptoms. The pathogenic bacterium was re-isolated from symptomatic plant tissue and confirmed as original isolates by 16S rRNA. Repetitive element sequence-based PCR and enterobacterial repetitive intergenic consensus PCR primers revealed different genetic profiles within P. syringae pathovars. High antibiotic susceptibility results showed the misreading of mRNA caused by streptomycin and oxytetracycline.

• Applied Biological Chemistry
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• v.43 no.3
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• pp.218-224
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• 2000

The efforts were made to optimite ethanol extraction from persimmon leaf with the time of extraction$(1.5{\sim}2.5\;hrs)$, the temperature of extraction$(70{\sim}90^{\circ}C)$, and the concentration of ethanol$(0{\sim}40%)$ as three primary variables together with several functional characteristics of persimmon leaf as reaction variables. The conditions of extraction was best fitted by using response surface methodology through the center synthesis plan, and the optimal conditions of extraction were established. The contents of soluble solid and soluble tannin went up as the concentration of ethanol went up and the temperature of extraction went down, and the turbidity went down as the concentration of ethanol went down. Electron donation ability was hardly affected by the extraction temperature and had the tendency to go up as the concentration of ethanol went up. The inhibitory activity of xanthine oxidase(XOase) had the tendency to go up as both the concentration of ethanol and the temperature of extraction went up. The inhibitory activity of angiotensin converting enzyme(ACE), the significance of which still was not recognized, showed the maximum when the concentration of ethanol was 27%. In result, the optimal conditions of extraction was the extraction time of two hours, the extraction temperature of $75{\sim}81^{\circ}C$, and the ethanol concentration of $33{\sim}35%$.

• Applied Biological Chemistry
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• v.43 no.4
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• pp.291-297
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• 2000

This study was conducted to find out the environmental factors affecting the differences in the half-life of the insecticide cyfluthrin in soil between field and laboratory tests carried out in 1998. Degradation and leaching of cyfluthrin in soil were examined under various environmental conditions that were considered to affect the residuality. Cyfluthrin was degraded 1.9 times faster in non-sterilized soil than in sterilized soil and 1.2 times at $25^{\circ}C$ than at $15^{\circ}C$. The half-lives of cyfluthrin were 61.4 days under the dark condition and 4.5 days under sunlight, and those were 11.8 days under the open condition and 23.8 days under the closed condition. The half-lives of the authentic compound and the commercial product of cyfluthrin were 15 and 1 day in the field test and 26 and 3 days in the laboratory test, respectively. Cyfluthrin was rapidly degraded with an increase in soil moisture content and decomposed faster in the alkaline solution of pH 12 than in the acidic solution of pH 3, but the half-life of cyfluthrin did not make any difference between pH 6.4 of the field test soil and pH 5.6 of the laboratory test soil. Cyfluthrin was immobile in soil from the results that $81{\sim}94%$ of the initial amount remained in the $0{\sim}2\;cm$ layer of the soil column regardless of the amount and time of rainfall after the chemical treatments. From viewing the abovementioned results, soil moisture content, sunlight and formulation type affected greatly soil microbes and volatilization affected slightly, and temperature, pH and rainfall did not affect the big difference in the half-life of cyfluthrin in soil between the field and laboratory tests in the year of 1998.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.519-526
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• 2008

To improve the quality of traditional kochujang, strains with high protease and amylase activity were isolated and identified from Sunchang traditional kochujang. Twenty-three strains strongly producing protease and 16 strains strongly producing $\alpha$- and $\beta$-amylase were isolated by using 1% isolated soy protein agar medium and 2% starch agar medium, respectively. Protease activities of the IA7, I5, and IA2 strain were 22.5, 21.2, and 20.6 unit/mL, respectively, and were higher than those of the other strains. Stains with high $\alpha$-amylase activity included K9 (967.8 unit/mL), K14 (828.3 unit/mL), K13 (662.5 unit/mL), K8 (601.5 unit/mL), and K11 (405.9 unit/mL). The $\beta$-amylase activity of the K11 strain was the highest, 34.3 unit/mL, among the isolated strains. Based on morphological, physiological properties, and API 50CHB-kit test for assimilation of 49 carbohydrates, 8 strains selected according to protease, $\alpha$-amylase, and $\beta$-amylase activities were tentatively identified as Bacillus megaterium (IA2), Bacillus subtilis (IA7, 15), Bacillus amyloliquefaciens (K8, K9, K11, and K13), and Bacillus stearothermophillus (K14). The IA7, 15, and K11 strains were finally identified as B. subtilis (99% ID) based on 16S rDNA sequencing.

• Animal cells and systems
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• v.15 no.1
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• pp.29-36
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• 2011

Insect lysozymes are basic, cationic proteins synthesized in fat body and hemocytes in response to bacterial infections and depolymerize the bacterial cell wall. The c-type lysozyme of the insect Spodoptera litura (SLLyz) is a single polypeptide chain of 121 residues with four disulfide bridges and 17 rare codons and is approximately 15 kDa. The full-length SLLyz cDNA is 1039 bp long with a poly(A) tail, and contains an open reading frame of 426 bp long (including the termination codon), flanked by a 54 bp long 5' UTR and a 559 bp long 3' UTR. As a host for the production of high-level recombinant proteins, E. coli is used most commonly because of its low cost and short generation time. However, the soluble expression of heterologous proteins in E. coli is not trivial, especially for disulfide-bonded proteins. In order to prevent inclusion body formation, GST was selected as a fusion partner to enhance the solubility of recombinant protein, and fused to the amplified products encoding mature SLLyz. The expression vector pGEX-4T-1/rSLLyz was then transformed into E. coli BL21(DE3)pLysS for soluble expression of rSLLyz, and the soluble fusion protein was purified successfully. Inhibition zone assay demonstrated that rSLLyz showed antibacterial activity against B. megaterium. These results demonstrate that the GST fusion expression system in E. coli described in this study is efficient and inexpensive in producing a disulfide-bonded rSLLyz in soluble, active form, and suggest that the insect lysozyme is an interesting system for future structural and functional studies.

• Journal of Microbiology and Biotechnology
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• v.26 no.10
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• pp.1708-1716
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• 2016

Glucose dehydrogenase (GDH) is an oxidoreductase enzyme and is used as a biocatalyst to regenerate NAD(P)H in reductase-mediated chiral synthesis reactions. In this study, the glucose 1-dehydrogenase B gene (gdhB) was cloned from Bacillus thuringiensis subsp. kurstaki, and wild-type (GDH-BT^WT) and His-tagged (GDH-BT^N-His, GDH-BT^C-His) enzymes were produced in Escherichia coli BL21 (DE3). All enzymes were produced in the soluble forms from E. coli. GDH-BT^WT and GDH-BT^N-His showed high specific enzymatic activities of 6.6 U/mg and 5.5 U/mg, respectively, whereas GDH-BT^C-His showed a very low specific enzymatic activity of 0.020 U/mg. These results suggest that the intact C-terminal carboxyl group is important for GDH-BT activity. GDH-BT^WT was stable up to 65℃, whereas GDH-BT^N-His and GDH-BT^C-His were stable up to 45℃. Gel permeation chromatography showed that GDH-BT^WT is a dimer, whereas GDH-BT^N-His and GDH-BT^C-His are monomeric. These results suggest that the intact N- and C-termini are required for GDH-BT to maintain thermostability and to form its dimer structure. The homology model of the GDH-BT^WT single subunit was constructed based on the crystal structure of Bacillus megaterium GDH (PDB ID 3AY6), showing that GDH-BT^WT has a Rossmann fold structure with its N- and C-termini located on the subunit surface, which suggests that His-tagging affected the native dimer structure. GDH-BT^WT and GDH-BT^N-His regenerated NADPH in a yeast reductase-mediated chiral synthesis reaction, suggesting that these enzymes can be used as catalysts in fine-chemical and pharmaceutical industries.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.7
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• pp.1053-1058
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• 2003

The solvent extracts of Houttuynia cordata root, which were extracted by using several solvents with different polarities, were prepared for utility as a natural preservatives. The antimicrobial activity was investigated by disc diffusion method against 22 microorganisms consisting of food borne pathogens, food poisioning microorganisms and food-related bacteria. The extraction yields were 15.7%, 3.7%, 0.13%, 0.5% and 5.9% in ethanol, chloroform ethylacetate, butanol and aqueous fractions, respectively. Antimicrobial activities were shown in ethanol, ethylacetate and butanol fraction of Houttuynia cordata root. However chloroform and aqueous fractions showed weak antimicrobial activity against the tested bacteria. Among the four fractions, ethylacetate fraction showed the strongest antimicrobial activities against microorganisms tested, such as B. megaterium, E. coli, K. pneumoniae and S. typhimurium. The polyphenolic compounds widely occuring in the traditional medicine plants have been reported to possess high antimicrobial activity. The polyphenolic compound in ethylacetate and butanol fraction were 35.9% and 16.0%, ethanol, chloroform and aqueous fraction were 5.0%, 2.3% and 1.7%, respectively. There are some relationship between antimicrobial activity and polyphenol content in natural plants. The ethylacetate fraction could be suitable for the development of a food preservative.