• Preventive Nutrition and Food Science
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• v.9 no.1
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• pp.14-20
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• 2004

Bacillus strains capable of producing fibrinolytic enzyme were isolated from traditional fermented Korean soybean paste and Japanese fermented soybean (Natto). Among the 16 strains, a selected Bacillus sp. was identified as bacillus firmus, with 80.7％ homology, by API kit analysis. Seed starter or B. firmus NA-1 was prepared with 5％ soymilk prepared from micronized soybean powder. To produce fibrinolytic enzyme by B. firmus NA-1 the liquid culture was performed with NB broth (pH 7.0) fortified with 1％ galactose, 0.1% tryptone, and 0.5% $K_2$HPO$_4$, by shaking with 180 rpm at 37$^{\circ}C$. Fibrinolytic enzyme activity reached the highest value at 7.8 unit/mL (plasmin unit) after fermentation for 72 hr. The crude fibrinolytic enzyme showed higher relative activity in the range of pH 7.0∼9.0. The activity of crude fibrinolytic enzyme was well maintained even after concentration by the vacuum evaporation at 5$0^{\circ}C$ for 1 hr.

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1143-1149
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• 2003

To elucidate a method of preventing dental caries, strains producing lytic enzymes were isolated and their characteristics were investigated. Among 5,00 alkalophilic strains isolated from soil, 22 strains showed lytic activity against Streptococcus mutans. Strain No. 4830, with the highest lytic activity, was selected for further study. Strain 4830 showed 94% sequence homology with the 16S rDNA sequence of Bacillus alcalophilus, but it was concluded to be different from Bacillus alcalophilus because of its biochemical characteristics. The strain was named Bacillus sp. 4830. The lytic enzyme from Bacillus sp. 4830 was purified by ethanol precipitation and CM-agarose column chromatography. The molecular weight of the lytic enzyme was determined to be 28 kDa by SDS-PAGE. The lytic enzyme was stable between pH 5.0 and pH 11 and up to $40^{\circ}C$. The optimal pH and temperature for the lytic activity was 9.0 and $50^{\circ}C$, respectively.

• Journal of Life Science
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• v.9 no.1
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• pp.111-120
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• 1999

A bacteriocin-producing strain, J-105, was isolated from Kimchi and identified as Lactococcus sp. The optimum conditions for the bacteriocin production from the isolated microorganism were evaluated. For maximum yield of bacteriocin from Lactotoccus sp. J-105, the cell should be harvested at the early stationary phase and temperature, pH and NaCl concentration should be $25^{\circ}C$, pH 8.0 and without the addition of NaCl, respectively. Maltose should be used as a carbon source and organic nitrogen such as polypeptone should be used as a nitrogen source for the best yield. The bacteriocin from isolate was inhibitory against Acetobacter aceti, Bacillus subtilis and several strains of lactic acid bacteria. The bacteriocin of J-105 was sensitive to pepsin, but stable for heat treatment. It was stable even at autoclaving temperature for 15 min.

• KSBB Journal
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• v.24 no.6
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• pp.508-514
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• 2009

This study investigated the production of catalase from Bul-kyo soil bacteria through fermentation process. Isolation and selection of bacteria was performed through chemical and physiological analysis. Catalases were produced from bacteria which belong to 3 different species (Bacillaceae bacterium BKBChE-1, Bacillus sp. BKBChE-2, Bacillus flexus BKBChE-3) confirmed by using 16S rDNA sequence method. The catalases were found to be stable in the temperature range of $30^{\circ}C-60^{\circ}C$ for BKBChE-1, BKBChE-2 and BKBChE-3 and also in the pH range of 9.0-12.0 for BKBChE-1 and BKBChE-3. Long-term stability of the catalases was about 20 days at $4^{\circ}C$. However, BKBChE-2 has kept its activity over 30 days at $4^{\circ}C$.

• Journal of Microbiology
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• v.40 no.1
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• pp.26-32
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• 2002

From the culture supernatant of the psychrotrophic strain of Bacillus amyloliquefaciens an extracellular serine protease was purified to apparent homogeneity by successive purification steps using QAE-Sephadex, SP-Sephadex and Sephacryl S-100 column chromatography. The pretense is monomeric, with a relative molecular mass of 23,000. It is inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride, but not by EDTA. The enzyme is most active at pH 9-10 and at $45^{\circ}C$, although it is unstable at $60^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.77-84
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• 2003

Chitinase was purified from an antagonistic bacterium Bacillus sp. 7079 by ammonium sulfate precipitation, QAE-Sephadex anion exchange chromatography, Sephadex G-100 gel filtration, and SP-Sephadex cation exchange chromatography. The molecula. weight of purified chitinase (PC-1) was approximately 66.5 kDa on SDS-PACE. PC-1 exhibited optimum pH and temperature of pH 7.5 and $45^{\circ}C$, respectively. More than $80\%$ of PC-1 was stable at pH 5.0 to 9.0, and more than $90\%$ at $40^{\circ}C$. $Fe^2+\;and\;Ca^2+$ inhibited the chitinase activity about $20\%$, and EDTA and p-CMB by about $30\%$, whereas $Ag^+$ inhibited the activity up to $65\%$. The $K_m$ value of PC-1 was 1.215 mg/ml with colloidal chitin as a substrate. We also investigated the effect of PC-1 treated chitin (PCTC) on the pro-inflammatory cytokine gene expression in macrophage RAW 264.7 cells. The expression of IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA gene was investigated using reverse transcriptase polymerase chain reaction (RT-PCR). IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were induced by the treatment of PCTC and chitin only in RAW 264.7 cells. These expressions were induced as early as 2 h and sustained up to 24 h in RAW 264.7 cells. IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were more strongly expressed by the treatment of PCTC than chitin treatment alone in RAW 264.7 cells.

• Journal of Microbiology and Biotechnology
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• v.21 no.6
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• pp.627-636
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• 2011

A commercially important alkaline protease, produced by Bacillus sp. RRM1 isolated from the red seaweed Kappaphycus alvarezii (Doty) Doty ex Silva, was first recognized and characterized in the present study. Identification of the isolated bacterium was done using both biochemical characterization as well as 16S rRNA gene sequencing. The bacterial strain, Bacillus sp. RRM1, produced a high level of protease using easily available, inexpensive agricultural residues solid-state fermentation (SSF). Among them, wheat bran was found to be the best substrate. Influences of process parameters such as moistening agents, moisture level, temperature, inoculum concentration, and co-carbon and co-nitrogen sources on the fermentation were also evaluated. Under optimized conditions, maximum protease production (i.e., 2081 U/g) was obtained from wheat bran, which is about 2-fold greater than the initial conditions. The protease enzyme was stable over a temperature range of 30-$60^{\circ}C$ and pH 6-12, with maximum activity at $50^{\circ}C$ and pH 9.0. Whereas the metal ions $Na^+$, $Ca^{2+}$, and $K^+$ enhanced the activity of the enzyme, others such as $Hg^{2+}$, $Cu^{2+}$, $Fe^{2+}$, $Co^{2+}$, and $Zn^{2+}$ had rendered negative effects. The activity of the enzyme was inhibited by EDTA and enhanced by $Cu^{2+}$ ions, thus indicating the nature of the enzyme as a metalloprotease. The enzyme showed extreme stability and activity even in the presence of detergents, surfactants, and organic solvents. Moreover, the present findings opened new vistas in the utilization of wheat bran, a cheap, abundantly available, and effective waste as a substrate for SSF.

• Microbiology and Biotechnology Letters
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• v.34 no.1
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• pp.7-14
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• 2006

Feather, generated in large quantities as a byproduct of commercial poultry processing, is almost pure keratin, which is not easily degradable by common professes. Four strains, SMMJ-2, FL-3, NO-4 and RM-12 were isolated from soil for production of extracellular keratinolytic protease. They were identified as Bacillus sp. based on their morphological and physiological characteristics. They shown high protease activity on 5.0% skim milk agar medium and produced a substrate like mucoid on keratin agar medium. Bacillus sp. SMMJ-2 had a faster production time for producing keratinolytic protease than other strains. This strain did not completely degrade whole chicken feather for five days in basal medium but completely degraded whole chicken feather when supplied with nitrogen source for 40hours in keratinolytic producing medium ($0.7%\;K_{2}HPO_{4},\;0.2%\;KH_{2}PO_{4},\;0.1%$ fructose, 1.2% whole chicken feather, $0.01%\;Na_{2}CO_3$, pH 7.0). When supplied with chicken feather as nitrogen source, keratinolytic protease activity was 89 units/ml/min. When soybean meal was used as nitrogen source, the keratinolytic protease production reached a maximum of 106 units/ml/min after 48 hours under $30^{\circ}C$, 180 agitation. To isolate the keratinolytic protease, the culture filtrate was precipitated with $(NH_4)_{2}SO_4$ and acetone. The recovery rate of keratinolytic protease was about 96% after treatment with 50% acetone. The enzyme was stable in the range of $30{\sim}50^{\circ}C$ and pH $6.0{\sim}12.0$.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.244-250
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• 2000

An ${\alpha}-glucosidase$ gene (aglA) from thermophilic Bacillus sp. DG0303 was cloned, sequenced, and expressed in Escherichia coli. The aglA was localized to the 2.1-kb PvuI-XmnI region within the 5.9-kb DNA insert of the gybrid plasmid pAG1. The gene consisted of an open reading frame of 1,686 bp with an unusual GTG initiation codon and TGA termination codon. The amino acid sequence deduced from the nucleotide sequence predicted a protein of 562 amino acid residues with a M, of 66,551 dalton. A comparative amino acid sequence analysis revealed that DG0303 ${\alpha}-glucosidase$ is related to bacillary oligo-1, 6-glucosidases. The Bacillus sp. DG0303 ${\alpha}-glucosidase$ showed a high sequence identity (36-59%) to the B. flavocaldarius, B. cereus, and B. thermoglucosidasius oligo-1, 6-glucosidases. The number of prolines in theses four ${\alpha}-glucosidases. was observed to increase with increasing thermostability of these enzymes. The cloned ${\alpha}-glucosidase was purified from E. coli $DH5{\alpha}$ bearing pAG1 and characterized. The recombinant enzyme was identical with the native enzyme in its optimum pH and in its molecular mass, estimated by sodium dodecy1 sulfate-polyacrylamide gel electrophoresis. The temperature optimum of the cloned ${\alpha}-glucosidase$ was lower than that of the native enzyme.

• Microbiology and Biotechnology Letters
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• v.27 no.5
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• pp.370-377
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• 1999

Thermo-tolerant bacterium producing the xylanase was isolated from soil and identified as Bacillus pumilus. This strain, named Bacillus pumilus TX703, was able to grow ad produce xylanase at the culture temperature of 5$0^{\circ}C$. The maximum xylanase production was obtained when 1%(w/v) birchwood xylan and 1% (w/v) soytone were used as carbon source and nitrogen source, respectively. The biosynthesis of xylanase was under the catabolite repression induced by glucose in the culture medium, and it was completely inhibited in the presence of 0.2% (w/v) glucose. The maximum activity of xylanase was observed from pH8.0 to 9.0 and from 50 to 6$0^{\circ}C$ and the enzyme was highly heat-stable, whose activity remained was over 50% at 8$0^{\circ}C$, and was quite stable from pH5.0 to 10.0.