• Applied Biological Chemistry
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• v.40 no.6
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• pp.495-500
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• 1997

To isolate a gene for cyclodextrin glycosyltransferase (CGTase) from alkalophilic Bacillus sp. E1, polymerase chain reaction (PCR) amplification was carried out. Direct molecular cloning of 1.2 kbp fragment and partial nucleotide sequence analysis of the PCR amplified clone, pH12, showed close homology with CGTases from Bacillus species. To investigate the genomic structure of the gene, Southern blot analysis of genomic DNA was carried out with the clone pH12 as a molecular probe. It showed that 5.3 kbp XbaI fragment was hybridized with the probe pH12. To isolate a genomic clone, genomic DNA library was constructed and a genomic clone for CGTase, pCGTE1, was isolated. Nucleotide sequence analysis of the clone pCGTE1 revealed that BCGTE1 contained 2,109 bp open reading frame encoding a polypeptide of 703 amino acids and showed over 94.3% amino acid sequence homology with CGTase of ${\beta}-cyclodextrin$ producer, Bacillus sp. KC201.(Received October 7, 1997; accepted October 20, 1997)

• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.384-389
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• 1992

Bacillus sp. YBL-7 which had been isolated from ginseng root-rot suppressive soil was able to antagonize Fusarium solani causing ginseng root-rot by their antibiotic substance. In order to develop multifunctional antagonist on Bacillus sp. YBL-7 as a biocontrol agent against Fusarium salam', optimal conditions for protoplast transformation system of Bacillus sp.YBL-7 by the vector plasmid pE194 were investigated. The protoplasts of Bacillus sp. YBL-7 were obtained at best efficiency by treatment with 200${\mu}g$/ml of lysozyme in the pH 7.0 of SMM buffer for 90 minutes at $40^{\circ}C$. The cell wall of the protoplast was regenerated on the agar plate containing 1.2% agar and 0.7 M mannitol. Under the best condition for protoplast formation and regeneration, the optimal transformation was achieved with 40% polyethylene glycol (M.W. 4000) treatment for 10minutes. The vector plasmid pE194 showed the best transformation frequency at 5$\mu$g/ml of final concentration. The pE194 was very stable over 80% in the transformants.

• Microbiology and Biotechnology Letters
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• v.17 no.6
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• pp.574-579
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• 1989

Chromosomal DNA fragments of Bacillus sp. YA-14, isolated from soil as a potent $\beta$-xylosidase producing bacterium, were ligated to a vector plasmid pBR322 and used to transfer Escherichia coli HB101 cells. The recombinant plasmid pYXL22 was found to enable the transformants to produce $\beta$-xylosidase. pYXL22 was found to contain the 7.0 kb HindIII DNA fragment originated from the Bacillus sp. YA-14 chromosomal DNA by Southern hybridization. The optimum temperature for the reaction of $\beta$-xylosidase produced by E. coli HB101 (pYXL22) was appeared at 3$0^{\circ}C$. The enzyme was maintained stably up to 4$0^{\circ}C$ when stored 1hr at 4$0^{\circ}C$. The $\beta$-xylosidase was repressed completely by 0.4% (w/v) glucose concentration in E. coli HB101 (pYXL22). The optimum concentration of xylose for the $\beta$-xylosidase production in Bacillus sp. YA-14 was 0.2% (w/v).

• Journal of Life Science
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• v.22 no.6
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• pp.799-806
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• 2012

Approximately 80 species of bacteria were isolated from the fermented mushroom first and the HJ-57 antibacterial micro-organism was selected to the final isolation bacteria. It has a high degree of CMCase, amylase, and protease activity as well as high antibacterial activity against mushroom pathogenic bacteria without affecting the growth and development of Flammulina velutipes and Lentinus edodes mushrooms. The finally selected HJ-57 antibacterial micro-organism was identified as Bacillus sp. HJ-57. The initial pH for culture was pH7 and its optimum culture temperature was $35^{\circ}C$. The antibacterial material produced by Bacillus sp. HJ-57 showed a little antibacterial activity even in the 12 hr of culture, but showed the highest antibacterial activity in the 36~48 hr of culture. The HJ-57 antibacterial micro-organism also showed a high antibacterial activity against mushroom pathogenic bacteria and molds in the corn cob contained culture medium is used in Flammulina velutipes cultivators.

• Journal of Microbiology and Biotechnology
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• v.3 no.3
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• pp.139-145
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• 1993

The nucleotide sequence of the xylanase gene (xynS) from alkali-tolerant Bacillus sp. YA.14 was determined and analyzed. A 639 base pairs open reading frame for xynS gene was observed and encoded for a protein of 213 amino acids with a molecular weight of 23, 339. S1 nuclease mapping showed that the transcription initiation site of the xynS gene did not exist in the cloned DNA. Ribosome binding site sequence with the free energy of -18.8 Kcal/mol was observed 8 base pairs upstream from the initiation codon, ATG. The proposed signal sequence consisted of 28 amino acids, of which 3 were basic amino acid residues and 21 were hydrophobic amino acid residues. When the amino acid sequences of xylanases were compared, Bacillus sp. YA-14 xylanase showed 48% homology with Bacillus sp. YC-335 xylanase and 96% homology with xylanases from B. subtilis and B. circulans.

• Journal of Life Science
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• v.20 no.2
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• pp.269-274
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• 2010

An antifungal antibiotic-producing strain, BCNU 2003, was isolated from forest soil in Korea. The morphological and physiological characters, and 16S rRNA sequences analysis of strain BCNU 2003 identified this strain as Bacillus genus. The Bacillus sp. BCNU 2003 showed strong antifungal activities against Aspergillus niger, Trichophyton mentagrophytes and Trichophyton rubrum with inhibition ranging from 62.05 to 63.49% by using dual culture technique. Bacillus sp. BCNU 2003 produced a maximum level of antifungal substances under aerobic incubation at 28oC and pH 6.5-7.2 for 6 days in LB broth. Ethyl acetate extract of the cultured broth showed strong antifungal activity and a broad antifungal spectrum against various pathogenic fungi. The minimum inhibitory concentration (MIC) values for its active extracts ranged between 0.0625 mg/ml and 1 mg/ml. In addition, Bacillus sp. BCNU 2003 was determined to have the ability to produce enzymes such as amylase, protease, gelatinase and catalase.

• Journal of Marine Bioscience and Biotechnology
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• v.1 no.4
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• pp.268-274
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• 2006

Bacillus sp. DYL130 producing biosurfactant was isolated from soil samples in the Duck-yu mountain and identified as Bacillus sp. by analysis of 16S rDNA sequence. Purification of the biosurfactant was performed by using affinity chromatography and TLC. The biosurfactant of culture medium from Bacillus sp. DYL130 was eluted with 100% methanol using affinity chromatography. To remove methanol, a rotary evaporator was used and enrichment sample was dissolved in alkaline water(pH 10). The purified biosurfactant was identified by TLC. It was confirmed that the Rf value of the biosurfactant was 0.78. Antifungal activity against Botrytis cineria was showed the strongly activity as active antagonist. Maximum emulsification activity and stability were obtained from soybean oil. The critical micelle concentration (CMC) of purified biosurfactant was 35mg/l and the purified biosurfactant inhibited biofilm forming by Bacillus sp..

• BMB Reports
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• v.28 no.4
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• pp.348-352
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• 1995

A thermophilic Bacillus sp. strain KK-1 isolated from soil produced an extracellular xylanase. From the culture supernatant of Bacillus sp., the xylanase was purified to homogeneity by ammonium sulfate precipitation and DEAE-Sephadex A-50 chromatography. The molecular weight of the purified xylanase was estimated to be 45 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel permeation chromatography. The apparent $K_m$ values for xylanase, using oat spelt xylan and birchwood xylan as substrates, were 7.1 mg/ml and 3.2 mg/ml, and $V_{max}$ values were $27.0\;{\mu}mol{\cdot}min^{-1}{\cdot}mg^{-1}$ and $29.0\;{\mu}mol{\cdot}min^{-1}{\cdot}mg^{-1}$, respectively. The xylanase hydrolyzed oat spelt xylan to mostly xylobiose, xylotriose, and xylose. The amino acid composition indicated that the xylanase contained high amounts of amino add residues of glutamic acid and glutamine (Glx) and aspartic acid and asparagine (Asx).

• Microbiology and Biotechnology Letters
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• v.29 no.2
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• pp.110-114
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• 2001

Isolation of BacilLus sp. producing multi-enzyme and optimization of medium conditions for its production using feedstuffs for probiotics were carried out in this study. A bacterium isolated from natural resources, namely Bacillus subtilis 4-3, has multi-enzyme activity (phytase. cellulase, xylanasc, protease, and amylase. In the culture of B. subtilis 4-3 using soybean meal and rice bran. relatively low phytate degradation was noted using whereas high phytate degradability was observed with wheat bran (80.63%). The optimal composition of medium using feedstuffs was 1.0% (w/v) soybean meal and 2% (w/v) molasses to yield high cell growth.

• Journal of Korea Soil Environment Society
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• v.4 no.2
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• pp.33-43
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• 1999

A strain (designated SK3l) which produces an excellent adsorption substance was isolated from soil samples and identified as Bacillus specied. The major adsorption substance (biosorbent SK3l) produced by Bacillus sp. SK31 was purified by ethanol precipitation and cetylpyridinium (CPC) precipitation. The adsorption charactics of zinc and lead ions on bioadsorbent SK3l were investigated. The equilibrium isotherms showed that bioadsorbent SK3l took up zinc and lead from aqueous solutions to the extent of about 52 mg/g and 112 mg/g. respectively. The culture conditions at the flask level of Bacillus sp. SK3l were investigated for the production of polysaccharide bioadsorbent, SK3l. The optimum pH and temperature for sorbent production were 7.5 and $30^{\circ}C$, respectively. The important carbon and nitrogen sources for sorbent formation were glucose and ammonium nitrate, respectively. In the optimized medium, sorbent production was improved three folds in comparison with the basal medium. In the jar fermenter, the highest sorbent production was obtained at 60 h cu1tivation time and the amount of biosorbent SK3l at that time was 9.2 g/$m\ell$.