Yong-Gyun Kim

Increased production of bacterial cellulose by Acetobacter sp. V6 in synthetic media under shaking culture conditions

Acetobacter strains are bacteria that can synthesize cellulose when grown in a complex medium containing glucose. The effect of the components of a synthetic medium on bacterial cellulose (BC) production by a newly isolated Acetobacter sp. V6 in shaking cultures was investigated. BC production was dependent on the presence of MgSO4 x 7H2O and cosubstrates such as ethanol and lactic acid in the medium. The optimal synthetic medium contained 1.5% glucose, 0.2% (NH4)2SO4, 0.3% KH2PO4, 0.3% Na2HPO4 x 12H2O, 0.08% MgSO4 x 7H2O, 0.0005% FeSO4 x 7H2O, 0.0003% H3BO3, 0.00005% nicotinamide, and 0.6% ethanol. A maximum BC concentration of 4.16 g/l was achieved after 8 days of cultivation at 200 rpm. The production of BC by Acetobacter sp. V6 was higher in synthetic medium than complex medium (Hestrin and Schramm medium) traditionally used for Acetobacter strains.

Optimization of fermentation conditions for the production of bacterial cellulose by a newly isolated Acetobacter sp. a9 in shaking cultures

The optimum fermentation conditions for the production of cellulose by a newly isolated Acetobacter sp. A9 were determined by shaken cultures. The strain was able to produce cellulose at 25–30 °C with a maximum at 30 °C. Cellulose production occurred at pH 4.5–7.5 with a maximum at pH 6.5. The improved medium composition was 4% (w/v) glucose, 0.1% (w/v) yeast extract, 0.7% (w/v) polypeptone and 0.8% (w/v) Na2HPO4·12H2O. Under these culture conditions, 3.8 g/l cellulose was produced after 7 days of cultivation, although this strain produced only 2.2 g/l in the standard medium. The addition of ethanol to the improved medium enhanced cellulose production: in an improved medium containing 1.4% (v/v) ethanol, cellulose production was 15.2 g/l, which was about four times higher than that without ethanol. Addition of ethanol was found to eliminate the spontaneous mutation of Acetobacter sp. A9.

Antimicrobial and Anticancer Activity of Korean Traditional Soy Sauce and Paste with Chopi

The fruits of Zanthoxylum piperitum are known as having various physiology vitality, and the abstraction ingredient of the pericarp is also known as having strong antibiotic activities against various bacteria. Therefore, this study was carried out to estimate the effect of physiology vitality when the abstraction ingredient of Z. piperitum was added in soy sauce(Chopi-kanjang) and soybean paste(Chopi-doenjang). For the antibiotic activity against the pathogens of sitotoxism such as Staphylococcus aureus, Salmonella typhimurium, Vibrio parahemolyticus, Escherichia coli 0157:H7, the extracts of the Chopi-kanjang was added 1%, 2%, 4% pericarp of Z. piperitum in the manufacturing process of soy source. According to the results, the growth of E. coli 0157:H7 and V. parahemolyticus were respectively inhibited as 70% and 50% by the Chopi-kanjang added 2% of the ingredient. For the antibiotic effects of the aforementioned Chopi-kanjang against Sal. typhimurium and Sta. aureus, the growth of those pathogens was also inhibited between 40% and 60% according to the manufacturing period of Chopi-kanjang. It was confirmed that the antibiotic activity using the mixture of the abstraction ingredient and Chopi-doenjang was lower than those of Chopi-kanjang. In order to estimate the anticancer activity using by caspase-3 activity, the mixture of the abstraction ingredient of the pericarp of Z. piperitum and Chopi-kanjang was treated to leukemia cells. According to the results, the activities of caspase-3 using the mixture added 1%, 2% and 4% of the abstraction ingredient were respectively increased as much as 4, 12, 15 times comparing with the control which was treated with the soy source only. It could be that the mixture of the abstraction ingredient of the pericarp of Z. piperitum and soy source induced apoptosis, and the mixture of the abstraction ingredient and soybean paste had no effect on the activity of caspase-3. In order to find out the death of the aforementioned cells caused by necrosis or apoptosis, DNA fragmentation in the cell was examined. U-937 cells showed apoptotic DNA fragmentation in the incubation with Chopi-kanjang extract.

Isolation and Characteristics of Bacteria Showing Biocontrol and Biofertilizing Activities

To develop multifunctional microbial inoculant, microorganisms with antagonistic activity and biofertilizing activity were screened. Pantoea agglomerans and Bacillus megaterium from our laboratory culture collection, and strain MF12 from soil near poultry farm in Miryang were selected. On the basis of morphological, physiological studies and 16S rDNA sequence analysis, isolate MF12 was identified as the Bacillus pumilis. Three strains were studied for insoluble phosphate solubilization, indole-3-acetic acid (IAA) and siderophore production, ammonification ability, hydrolytic enzyme production and antifungal activity against phytopathogenic fungi. P. agglomerans did not produce any visible clear zone on agar plate containing 0.5% as a sole phosphorus source. However, this strain could solubilize insoluble phosphate in liquid medium. All strains produced IAA ranged from depending on culture time and had ammonification ability. Among three strains, only P. agglomerans produced siderophore. P. agglomerans produced pectinase and lipase, B. megaterium produced amylase, protease and lipase while B. pumilis produced protease and lipase. P. agglomerans showed antifungal activities against phytopathogenic fungi, Fusarium oxysporum and Colletotrichum gloeosporioides. B. pumilis showed antifungal activities against Botrytis cinerea, Sclerotinia sclerotiorum and Phythium ultimum.

Optimal Condition to Produce Protease by Strain Separated from the Intestine of Reticulitermes speratus

We separated the bacteria showing protease activity from Reticulitermes speratus which is known as the only termite species in Korea. Then, we collected the best activated strain and studied the optimal culture condition for producing the enzyme. According to the results of observing morphological and physiological characteristics, and the type of 16S rRNA of the strain, it was identified as Serratia marcescens and named S. marcescens strain TM-3215. This strain showed the best activity under conditions of 0.8% (w/v) starch, 0.4% (w/v) peptone, 0.08% (w/v) MgSO₄?7H₂O, 30℃ and pH 8.0. After being cultivated under optimal conditions for 9 hr, the strain produced 19.8 U/ml of enzyme, an amount 1.8 times greater than the control.

Structural Identification of Antibiotics from Pseudomonas sp. RRj 228, a Antifungal Activity of Collectotrichum acutatum Causing Anthracnose on Pepper

Microorganisms near the plant rhizosphere usually inhabit the surface or the inside of the plant roots and have a direct effect on plant growth by secreting plant growth promoters or antagonistic materials which protect the root zone system from various pathogens. This study was carried out to identify and isolate the antagonistic materials after isolation of microorganisms showing high antagonistic activities, in hopes of contributing to the development of sustainable agriculture and the preservation of agricultural environments. A number of antagonistic bacteria were isolated from paddy soil. Among isolates, RRj 228 showed plant growth promotion and antagonistic activity. RRj 228 was identified as Pseudomonas sp. according to the results of physiological properties and genetic methods. On the basis of the results of anti-fungal spectrum against several pathogens by RRj 228, the antagonistic effect of the isolate against Botrytis cinerea, Pythium ultimum, Phytopthola capsici, and Rhizoctonia solani, especially against red-pepper anthracnose caused by Colletotrichum acutatum, was remarkable. The experiment evaluating the biological control effect by RRj 228 revealed that the value by the RRj 228 culture against C. acutatum, R. solani and P. ultimum were 0.14 mg/ml, 0.16 mg/ml and 0.29 mg/ml, respectively. An antagonistic substance was isolated and purified by several chromatographies from the RRj 228 culture. The and assignment of the antagonistic substance was achieved from two-dimensional COSY, HMQC, and HMBC. Finally, the antagonistic substance was identified as Phenazine-1-carboxylic acid (, M.W.

Pretreatment of Cane Molasses for Production of Bacterial Cellulose and Its Physico-Chemical Properties

The aim of this study is to investigate cane molasses pretreatments for the production of cellulose by Acetobacter sp. V6, which has excellent bacterial cellulose (BC) producing capacity in the shaking culture. Among pretreatments of cane molasses, 1% (w/v) tricalcium phosphate (TP) treatment was more efficient in BC production. The physico-chemical properties of BCs that were produced in static and shaking cultures were also investigated. Although BC had an emulsifying ability, its emulsion stability was low. Water holding capacity (WHC) of BC was high; the WHC of BC produced in static culture was 14 times higher than that of α-cellulose. In addition, the viscosity of BC was higher than that of α-cellulose. Composition analysis by FT-IR showed no difference in composition between BC and plant cellulose. In the crystallinity analysis by XRD, all BC samples showed crystallinity. All BC samples showed reticulated structures consisting of ultrafine cellulose fibriles. Microfibriles of cellulose from static culture were especially more compact than those of cellulose from shaking culture.

Control effect of the newly developed insecticidal protectant on Sycamore lace bug, Corythucha ciliata (Hemiptera : Tingidae)

To determine the effects of newly developed insecticide (Ultra) on Sycamore lace bug (Corythucha ciliata), ten leaf discs were sprayed with the Ultra solution including the other selected commercialized insecticide solutions such as Carbaryl and Pirimicarb, and also with a water-sprayed control. According to the results, the control effect of Ultra on the adults and immatures was respectively 98.9% and 97.5% after 48hr, and the corresponding values were higher than that of the other tested insecticides (i.e Carbaryl showed 84.6% for adults and 85.6% for immatures; Pirimicarb showed 83.0% for adults and 90.7% for immatures). For the Susceptibility test, LC value of Ultra showed the lowest as 1.5ppm, the value of Carbaryl and Pirmicarb were 3.7 ppm and 30.4 ppm, respectively. Therefore, the insecticidal effects were better for the newly developed insecticide than the commercialized insecticides.

Amount of Telomeric DNA on Lymphocytes in Senescence Mouse by Quantitative Fluorescence in situ Hybridization

Telomeres, comprised of tandem repeats of TTAGGG sequences, are special nucleoprotein structures that protect and stabilize chromosome ends. These structures form the crux of the telomere concept of aging, senescence and genomic instability. The classic terminal restriction fragment (TRF) analysis to quantify the amount of telomeric DNA is disadvantageous in species containing ultra long telomeres like in mice (100Kb). In this study, we used a more sensitive quantitative fluorescence in situ hybridization (Q FISH) technique to quantify telomeric DNA, and used it as a biological aging marker in mice. 12 litters each of Senescence-Resistant (SAMR1) and ?Prone (SAMP1) known as senescence accelerated mouse strains were purchased from Central Lab, Animal Inc. We quantified the amount of telomeric DNA using telomere specific DNA probes on the two strains of male mice at 8 weeks, 18 weeks and 26 weeks of age. The amount of telomeric DNA correlated with aging and age associated changes in body and organ weight between SAMR1 and SAMP1 strains of mice. These data suggest the usefulness of the amount of telomeric DNA as a biological aging marker in human aging studies.

Partial Characterization and Induction of Ferulic Acid Esterase and Xylanase from Pseudomonas sp. LG2

Lignin degrading bacterium Pseudomonas sp. LG2 was able to degrade lignin substrate to a lot of APPL compound. APPL compound was detected in culture supernatants from Pseudomonas sp. LG2 grown with BSC(brewer`s spent grain). FAE(ferulic acid esterase) and xylanase are induced from Pseudomonas sp. LG2 in the presence of carbon sources such as oat spelt xylan, HBSG I, II(hydrolyzed brewer`s spent grain I, II) and AFBSG(autoclaved fraction from brewer`s spent grain). However, xylanase and FAE are not induced by growth of Pseudomonas sp. LG2 on xylose and arabinose. Pseudomonas sp. LG2 is grown on medium containing oat spelt xylan, HBSG I, II and AFBSG and the induction of FAE and xylanase activities of extracellular proteins determined during 14 days. Maximum level of xylanase activity(5.3 U/mg) found at 6 days in culture contained oat spelt xylan as carbon source, whereas maximum level of FAE activity(15.4 mU/mg) was found at 8 days in culture contained AFBSG as carbon source. Most ferulic acid was released in culture supernatants when Pseudomonas sp. LG2 grown on oat spelt xylan, HBSG I, II and AFBSG. FAE of extracellular enzymes was also specific activity on methyl ferulic acid, methyl caffeic acid and methyl p-coumaric acid respectively, but not methyl sinapinic acid, methyl vanillic acid and methyl gallic acid.