• KSBB Journal
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• v.13 no.6
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• pp.644-648
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• 1998

In the production of a low cost bacterial insecticide, it is important to produce a high spore concentration using low price substrates. Experiments were carried out to investigate the effects of the addition of mineral salts and glucose, and of dissolved oxygen concentration on the cell growth and spore formation of Bacillus thuringiensis var aizawai using a cheap wheat and soybean meal in the batch culture. The maximum viable cell number was 1.2${\times}$109 CFU/mL at 12 hr culture and spore yield was 54.2% at 74 hr culture using an industrial medium containing 20 g/L wheat meal and 30 g/L soybean meal under 1.0 vvm aeration and 200 rpm agitation. The cell growth and the spore formation were not enhanced by the addition of mineral salts in industrial medium, whereas th addition of 10g/L glucose decreased the cell growth and spore formation. We could obtain a maximum viable cell number of 2.2${\times}$109 CFU/mL and spore number of 1.9${\times}$109 CFU/mL at the dissolved oxygen concentration of 60% of saturation. The spore concentration was enhanced approximately by 2 times as compared to the dissolved oxygen concentration of 50%. In the bench-scale culture, the maximum viable cell and spore number were 2.5${\times}$109 CFU/mL, and 2.2${\times}$109 CFU/mL, respectively under 1.0 vvm aeration and 400 rpm agitation. The spore yield was 88% based on the maximum viable cell number. As a result, it was confirmed that the production of high spore concentration could be obtained by a bench-scale culture using an industrial medium.

• KSBB Journal
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• v.15 no.3
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• pp.219-225
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• 2000

Both the production of high spore concentration and high insecticidal activity are required in the production of Bacillus thuringiensis to be used for the bacterial insecticide. In the production of high cell and spore concentrations of B. thuringiensis the continuous fed-batch culture(CFBC) and intermittent fed-batch culture(IFBC) were investigated at $28^{\circ}C$ by maintaining 40% dissolved oxygen concentration. When the final glucose concentration was 50 g/L the maximum viable cell number obtained using the CFBC with linear gradient feeding was $9.37{\times}109$ cells/mL and maximum spore concentration was $8.33{\times}109$ spores/mL which was approximately 84.4% yield of spore formation. When the final glucose concentration was 100 g/L the aximum viable cell and spore concentrations obtained using the IFBC with pH-statb were $1.38{\times}$1010 cells/mL and $1.35{\times}1010$ spores/mL respectively and the yield of spore formation was approximately 97.8%.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.522.2-522
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• 1986

In order to optimally induce sporulation and toxin formation in Bacillus thuringiensis, exhaustion of specific nutrients as well as resuspension experiments were tried. Sporulation and toxin formation was most abunduntly occurred when the growth was limited by carbon source. It was also occurred in a resuspension medium containing only distilled water. Various environmental and physiological factors affecting the efficiencies of spore and toxin formation were examined in chemically defined media. As a result of these studies, a batch fermentation resulted in higher spore and toxin yield than ever reported

• Research in Plant Disease
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• v.8 no.2
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• pp.124-130
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• 2002

In 1998, a pathogen isolated from infected onions was identified as Aspergillus niger. At 3$0^{\circ}C$, A. niger AnYD-1 showed the best mycelial growth, spore germination, and high pathogenicity to onions. In spite of the weak inhibitory effect of flusilazole and hexaconazole on the mycelial growth on PDA, they showed the specific inhibitory activity against the formation of spores in the vapour phase. With flusilazole and hexaconazole, the effects of the solvent, the applied concentration and the incubating temperature on the activities inhibiting the spore formation were confirmed. Their inhibitory effect on the spore formation in vapour phase was excellent by solving them with dimethylsulfoxide and dimethylformamide among tested solvents, and applying them at high temperature such as 30~35$^{\circ}C$.

• KSBB Journal
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• v.26 no.3
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• pp.243-247
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• 2011

Using Bacillus subtilis spore display system, with cotG as an anchoring motif, levansucrase from Zymomonas mobilis, was displayed on the outer surface of Bacillus subtilis spore. Flow cytometry of DB104 (pSDJH-cotG-levU) spore, proved the surface localization of CotG-LevU fusion protein on the spore compared to that of DB104. Enzymatic activity of DB104 (pSDJH-cotG-levU) spore showed more than 1.5 times higher levansucrase specific activity compared to that of the host spore, which is a remarkable increase of enzymatic activity considering the existence of sacA (sucrase) and sacB (levansucrase) in the Bacillus subtilis chromosome. The spore integrity, revealed by sporulation frequency test after heat and lysozyme treatment of spore, did not changed at all in spite of the CotG-LevU fusion protein incorporation into the spore coat layer during spore formation process. These data prove again that Bacillus subtilis spore could be considered as good live immobilization vehicle for efficient bioconversion process.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.72-78
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• 2001

It is possible ot prepare protoplasts of the zygomycete fungus, Phycomyces blakesleeanus, by digesting the cell wall of spore germlings with commercially available chitinase and chitosanase. However, the cells without any cell walls immediately form large aggregates, and thus, it is difficult to isolate the individually separated protoplasts. Inherent problem with the formation of aggregates in preparing protoplasts could be solved by the use of bovine serum albumin (BSA). As a result, we were able to prepare a large number of single protoplsts quickly and easily. We took time-lapse photomicrographs of the formation of protoplasts, and found that there were certain regions of the cell wall of spore germlings that were sensitive to chitinase and chitosanase, although the cell wall of the original spores is known to be insensitive to these enzymes. There are two kinds of cell walls on a spore germling; one with a bound wheat germ agglutinin (WGA), and the other a bound concanavalin A (ConA). Furthermore, only cells with walls which had bound WGA were able to regenerate, while those with walls with bound ConA were not able to regenerate.

• Korean Journal of Microbiology
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• v.6 no.4
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• pp.113-121
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• 1968

This study has been aimed to determine the ecological factors with relationship to the Jongkok production in view of fermentation technology by means of some strains, Asp. kawachii, which is now preserved by the author and the following factors are included during the study; inorganic salts, nitrogen, sugar, water contents and temperature. The results, are as follows: (a) Sugar among other above-mentioned factors is increasingly affecting the number of the short type of conidiophore on culture medium and the conidiophore is increased by direct ratio until glucose concentration of 50%, at which concentration is mostly effective for the short type of conidiophore, while other factors did not affect on it. (b) Until glucose concentration of 50% sugar component of culture medium is favorable for the spore formation of Asp. kawachii by direct ratio. And peptone or asparagine on nitrogen medium, calcium-phosphate among other inorganic salts, wheat bran and rice branare also favorable, but other factors rientioned earlier show no relationship with the spore formation. Sugar, however, also related with the spore color clearness of crimson and light brown, and spore color is mostly clear at the point of glucose concentration until 50%. And asparagine on nitrogen medium, calcium phosphate among other inorganic salts, rice bran did all affect on the color clearness, while other factors did not concern with color clearness. (c) Water, sugar and temperature have related with the acid formation which is promoted, by direct ratio at the point of water-saturated condition and glucose concentration of 50%, while temperature at $25^{\circ}C$favorably affected on the acid formation which is increased by inverse ratio at the temperature$25^{\circ}C$ to $45^{\circ}C$ And pH did not relate with the acid formation. (d) Cylindrical plate method devised by the author is mostly favorable for the preservation and isolation of culture, compared with the traditional slant medium method.

• Mycobiology
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• v.33 no.4
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• pp.194-199
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• 2005

Application of fertilizers such as urea, diammonium phosphate (DAP) and muriate of potash in soil adversely affected the spore germination of Arthrobotrys dactyloides. Amendment of soil with urea at the concentrations of 1.0%, 0.5% and 0.1 % completely inhibited spore germination and direct trap formation on the conidium, whereas muriate of potash delayed and reduced the spore germination even at the lowest concentration. DAP also inhibited spore germination at 1.0% concentration, while at lower concentration the percentage of spore germination was reduced. Application of neem cake at the concentration of 0.5% also inhibited spore germination after 24 h of amendment. The inhibitory effect of neem cake was reduced after 15 days of amendment, while after 30 days after amendment the inhibitory effect was completely lost and the spore germinated by direct trap as in unamended soil. Nematodes were not attracted to ungerminated spores after 24 h of amendment. After 15 days of amendment nematodes were attracted to agar blocks containing fewer germinated spores after 24 h of incubation but after 48 h of incubation large number of nematodes were attracted and trapped by the germinated spores with direct traps. After 30 days of amendment, larger number of nematodes were attracted and trapped by direct traps.

• The Korean Journal of Mycology
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• v.12 no.4
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• pp.141-152
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• 1984

Aspergillus niger van Tieghem was cultured by the method of synchronous and submerged culture. Throughout the culture, sporulation was occured. Highly phosphorylated nucleotides in sporulating mycelia were detected to assure whether the eucaryotic Aspergillus niger produce these substances or not during the differentiation. Phosphorylated nucleotides were extracted from the conidiophore, bearing mycelia and spore forming body, these nucleotides were identified by TLC with P.E.I. cellulose. Guanosine tetraphosphate was found in both phialide forming mycelia and spore forming body. The contents of free amino acids were assayed and its level was found to increase at early stage of sporulation. The effects of 8-azaguanine examined, it was found to prevent spore formation and to made abnormal structure. The effects of inosinic monophosphate and guanosine monophosphate on spore formation were examined, spore formation was enhanced by these nucleotides.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.335-337
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• 2006

Glycerol-spore formation of wild myxobacteria isolated in Korea has been tested. One third of the wild myxobacterial isolates belonged to the genus Myxococcus and Corallococcus produced a large number of glycerol-spores in the presence of glycerol or dimethyl sulfoxide. However, two third of the other wild myxobacterial isolates belonged to the genus Myxococcus and Corallococcus produced a negligible number of spores under the same condition. In addition, the myxobacteral isolates belonged to the other genus, such as Stigmatella, Archangium, and Cystobacter, failed to produce a noticeable number of glycerol-spores.