• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.275-280
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• 2000

The inulin fructotransferse (depolymerizing) (IFTase, EC 2.4.1.93) gene of Arthrobacter sp. A-6 was cloned and expressed in Escherichia coli and Bacillus subtilis. The IFTase gene consisted of an ORF of 1.311 nucleotides encoding a polypeptide of 436 amino acids containing a signal peptide of 31 amino acids in the N-terminus. The molecular mass of the IFTase based on the nucleotide sequence was calculated to be 46.116 Da. The recombinant E. coli $DH5{\alpha}$ cells expressing the Arthrobacter sp. A-6 IFTase gene produced most of the IFTase intracelularly. In contrast, the recombinant B. subtilis DB 104 carrying the IFTas gene on a B. subtilis-E. Coli expression vector secreted the IFTase into the culture fluid efficiently.

• Journal of Microbiology and Biotechnology
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• v.8 no.1
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• pp.28-33
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• 1998

A gene coding for ${\beta}$-xylosidase from thermophilic xylanolytic Bacillus sp. KK-1 was cloned into Escherichia coli using plasmid pBR322. Recombinant plasmid DNAs were isloated from E. coli clones which were capable of hydrolyzing 4-methylumbelliferyl-${\beta}$-D xylopyranoside. Restriction analysis showed the DNAs to share a common insert DNA. Xylo-oligosaccharides, including xylotriose, xylotetraose, xylopentaose, and xylobiose were hydrolyzed to form xylose as an end product by cell-free extracts of the E. coli clones, confirming that the cloned gene from strain KK-1 is ${\beta}$-xylosidase gene. The ${\beta}$-xylosidase gene of strain KK-1 designated as xylB was completely sequenced. The xylB gene consisted of an open reading frame of 1,602 nucleotides encoding a polypeptide of 533 amino acid residues, and a TGA stop codon. The 3' flanking region contained one stem-loop structure which may be involved in transcriptional termination. The deduced amino acid sequence of the KK-1 ${\beta}$-xylosidase was highly homologous to the ${\beta}$-xylosidases of Bacillus subtilis and Bacillus pumilus, but it showed no similarity to a thermostable ${\beta}$-xylosidase from Bacillus stearothermophilus.

• Microbiology and Biotechnology Letters
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• v.34 no.1
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• pp.40-46
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• 2006

A bacterial strain producing high level of a phytase was isolated from cattle feces and identified as Bacillus subtilis, and designated as Bacillus sp. CF 5-26. The production of the phytase from Bacillus sp. CF 5-26 reached the highest level after 72 hours at $37^{\circ}C$. The optimum condition of the media for the production of phytase was 10% rice bran extract, 0.1% whey protein powder, $0.01%\;CaCl_{2},\;0.01%\;KH_{2}PO_4$. The phytase was purified 20.3 folds with ethanol precipitation, Sephadex G-100, CM Sepharose CL-6B and Sephacryl S-100-HR column chromatography. The molecular weight of the purified enzyme was estimated to be 66 kDa on SDS-polyacrylamide gel electrophoresis. The purified phytase activity was stable up pH 5.0, 7.0, 11.0 and the remaining activity was 50% when it was treated at $100^{\circ}C$ for 1 hour. The substrate specificity of phytase was most active against sodium phytate and inositol polyphosphate compound. And the phytase hydrolysed tripolyphosphate and pyrophosphate a little. The Km value for the sodium phytate was 0.64 mM and the Vmax value was $4.41\;{\mu}mol/min$.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.283-288
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• 1989

To reduce the size of 5.0kb HindIII fragment containing $\beta$-xylosidase gene, the 5.0kb insert of pAX278 which was previously cloned was reduced by various deletions and thus 1.4kb EcoRI-Xbal fragment was subcloned into pUC19, and the recombinant plasmid was named pAK208. The $\beta$-xylosidase acnivity of E. coli harboring pAK208 was higher about 1.3times than that of pAX278. For the improvement of $\beta$-xylosidase activity, we cloned and expressed the $\beta$-xylosidase gene in E. coli using vector pKK223-3 containing a potent tac-promoter, and enzyme activity of the transformant harboring pKHR212 was increased about 3.3 and 1.8 times than that of E. coli(pAX278) and Bacillus sp. K-17, respectively. To obtain better expression of $\beta$-xylosidase gene, the whole 5.0kb HineIII fragment was recloned into pC194, and the Bacillus sp. K-17 transformant harbor-ing the recombinant plasmid pCX174 showed higher activity than that of the E. coli (pAX278) and Bacillus sp. K-17, respectively. The characteristics of enzyme purified from transformants were consistent with those front alkalophilic Bacillus sp, K-17.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.318-322
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• 2006

The intracellular invertase gene of alkalophilic and thermophilic Bacillus sp. TA-11 which was isolated from compost was cloned and expressed in E. coil HB101 using pUC19 as a vector. The invertase of the recombinant E. coli (pYC 17) was maximally produced when it was incubated at 37$^{\circ}C$ for 9 h in a SY medium containing 0.25% sucrose, 0.5% yeast extract, 0.1% each of $K_2HPO_4$ and $KH_2PO_4$, with an initial pH of 8.0. The final enzyme activity under the above condition was 47.7 U per ml of cell-free extract.

• Microbiology and Biotechnology Letters
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• v.28 no.6
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• pp.329-333
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• 2000

Alkaline bacterium producing a high pro-tease activity at low temperature was isolated by using enrichment culture from soil samples and identified as Bacil-lus sp. WRD-1 Cell growth was maximal at 10 hours and the optimal initial pH and culture time of culture condition for enzyme production was pH 7 and 10 hours, respectively. Temperature range of high enzyme activity were $10~40^{\circ}C$. The optimal pH and temperature for the enzyme activity were pH9 and $30^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.25 no.5
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• pp.506-511
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• 1997

For the production of benzoylformic acid from DL-phenylglycine, 8 strains of bacteria capable of producing benzoylformic acid were isolated from soil. Among them, the strain YH-001 showed the highest activity for production of benzoylformic acid, and was partially identified as a Bacillus sp. For the production of benzoylformic acid, a reaction mixture and reaction conditions were optimized as follows: the reaction mixture contained 2 g of DL-phenylglycine and 50 g of wet cells in 1 l of 50 mM potassium phosphate buffer (pH 8.0) and the reaction was carried out at 40$circ$C with shaking. After 6 hr incubation, 1.01 g/l of benzoylformic acid was produced which coffesponded to a conversion yield of 50.5%.

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.156-163
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• 1992

Bacillus sp. was isolated from soil for its strong activity of cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19). The enzyme was purified by gel filtration and anion exchange column chromatography using FPLC. The purified enzyme exhibited its maximum CGTase activity in the pH range of 6~8 and the temperature range of 50~$70^{\circ}C$. The molecular weight was estimated as 114,000 by SDS-PAGE. The isoelectric point of the enzyme was 4.3. The CGTase of Bacillus sp. E l produced $\beta$-cyclodextrin mainly and did not produce a-cyclodextrin. The product ratio of $\beta$-cyclodextrin to $\gamma$-cyclodextrin was 7:l.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.44-48
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• 1990

Alkalophilic sp. YC-335 isolated from soil was capable of producing large amount of cyclodextrin glycosyltransferase (CGTase) in culture broth. This enzyme was successively purified 52.9 folds with 17.8 yield by ethanol precipitation, DEAE-Toyopearl column chromatography and Sephadex G-100 column chromatography. The purified enzyme have a molecular weight of approximately 75,000 estimated by SDS polyaerylamide gel electrophoresis. The optimum pH and temperature for the enzyme activity were 6.0 and 5$0^{\circ}C$, respectively. The enzyme stable between pH 6 and 10, and up to 5$0^{\circ}C$. The thermostability of the enzyme was increased up to 6$0^{\circ}C$ by the addition of 15mM CaCl$_2$.

• Journal of Life Science
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• v.9 no.1
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• pp.99-105
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• 1999

The aims of this studies were carried out to investigate the antioxidant activity on low density lipoprotein(LDL) using substances extracted from Bacillus sp. RH-5. The antioxidative substances produced extracellular in the culture broth by Bacillius sp. RH-S was obtained by elution of chloroform : methanol from silicagel column (80cm x100cm) chromatography. Band 4 eluted from fraction 3 by TLC was appeared at highest level of antioxidative activity using thiocyanate methed. Band 4 at a concentration of 100 or 200$\mu$g/$m\ell$ inhibited oxidation of LDL induced by the mouse transformed macrophage. According to IR. NMR or GC/MASS, the antioxidant substance was identified as 5-hydroxyindole.