• Applied Biological Chemistry
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• v.51 no.4
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• pp.299-304
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• 2008

A ${\beta}$-1,4-endoglucanase gene from Bacillus subtilis H12 was cloned into Escherichia coli JM109 (pBC8) and sequenced. The endoglucanase gene with an insert DNA of 2.5 kb possessed an open reading frame of 1,500 bp encoding a mature protein of 499 amino acids with a calculated molecular mass of 55 kDa. The deduced amino acid sequence showed similarity to those of the known neutral cellulase genes of B. subtilis PAP115 (99.2%) and BSE616 (97.8%), as well as the alkaline gene of Bacillus sp. N4 (55.1%). The endoglucanase activity expressed by E. coli (pBC8) was localized in the periplasmic fraction (80%) and the cytoplasmic fraction (20%). An endoglucanase was purified from the periplasmic fraction by performing gel filtration and anion exchange chromatography. The molecular weight of the purified enzyme was estimated to be 31 kDa by SDS-PAGE, and the maximum activity occurred at pH 7 and $40^{\circ}C$. The enzyme easily hydrolyzed soluble substrates such as carboxymethyl cellulose and barely ${\beta}$-glucan, whereas the sigmacell and xylan, the known insoluble substrates, were not entirely hydrolyzed.

• Journal of Applied Biological Chemistry
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• v.54 no.1
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• pp.7-14
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• 2011

Bacillus subtilis JK-1 showed degradation activity against crude oil, gasoline, kerosene, and light oil, and this strain was used as a crude biosurfactant producing microorganism in this study. To optimize the culture medium for production of crude biosurfactant, the influences of various carbon, nitrogen and mineral sources were assessed. The highest biosurfactant production by B. subtilis JK-1 was observed after 96 h cultivation, containing 1.0% (w/v) soluble starch as a carbon source and 0.5% (w/v) skim milk as a nitrogen source, and carbon to nitrogen concentraion (C/N) ratio was 2.0. For the biosurfactant production 0.1% (w/v) of $KNO_3$ was the most effective mineral source. Comparison of biosurfactant production indicates that B. subtilis JK-1 produces more biosurfactant in the optimum medium established in this study than LB and TSB. Under the optimum medium, the surface tension of culture broth of B. subtilis JK-1 was decreased from 47.3 dyne/cm to 24.0 dyne/cm after cultivation of 48 h.

• The Korean Journal of Food And Nutrition
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• v.11 no.4
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• pp.381-387
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• 1998

The effects of culture conditions on the production of amylase expressed by Bacillus subtilis LKS88 with a cloned gene from Streptomyces albus KSM-35 were investigated. The production of amylase was increased significantly by using sodium citrate and rice hull as a carbon source. In addition, the use of a mixture of sodium citrate and rice hull (1:1) resulted in increase of enzyme production by 20-fold when compared to that of soluble starch. The soybean meal as the nitrogen source could be partially replaced with yeast extract without changing the enzyme production yield. The amylase production was also increased by adjusting initial pH to 6.0 or by adding 0.01% SDS. Maximum amylase production was observed in the medium containing 1.5% sodium cirtate, 1.5% rice hull, 0.7% soybean meal, 0.3% yeast extract, 0.66% K2HPO4, 0.05% MgSO4$.$7H2O, 0.008% CaCl2$.$2H2O, 0.01% SDS with initial pH of 6.0. The maximum yield of amylase reached 56.6 U/ml when B. subtilits LKS88 (pASA 240) was cultured at 37$^{\circ}C$ for 36 hr.

• Microbiology and Biotechnology Letters
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• v.17 no.2
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• pp.160-164
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• 1989

A 5.7Kb EcoRI fragment containing alkaline amylase gene of Bacillus sp. AL-8 obtained in the previons experiment (10) was transformed in B. subtilis via plasmid pUB110. The enzymatic proper-ties of the amylase produced by the transformants were Identical to those of the donor strain. Thus, the alkaline amylase activity from the transformant was maximum at pH 10 and 5$0^{\circ}C$. And the enzyme was very stable over the ranges of alkaline pH. In order to determine the location of the alkaline amylase gene within the 5.7Kb DNA fragment, the fragment was subcloned in E. coli. It was found that the alkaline amylase gene was located k EcoRI fragment of 3.7Kb.

• Food Science and Biotechnology
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• v.18 no.4
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• pp.1035-1037
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• 2009

In this study we analyzed the dynamic changes in microbiota composition during kochujang fermentation at $30^{\circ}C$. During fermentation, the viable cell counts slowly increased and reached $3.2{\times}10^7$ for aerobic bacteria, $8.3{\times}10^3$ for yeast, and $1.4{\times}10^3$ CFU/mL for fungi after 60 days. Bacilli were found to be the most dominant microorganisms throughout the fermentation process. Using the culture dependent method Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloquefaciens were found to be the main species during the early stages of fermentation; however, Bacillus pumilus and Bacillus stearothermophilus became the most dominant species during the late stage of fermentation. In contrast, when the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method was used Bacillus ehimensis was found to be the dominant species during the early stage of fermentation and Bacillus megaterium, B. pumilus, B. subtilis, and B. licheniformis were dominant in the ate stages. These results indicate various other Bacillus species rather than just B. subtilis and B. licheniformis might be involved in the fermentation of kochujang.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1092-1100
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• 2012

Ten Bacillus strains with antimicrobial activities were isolated from Cheonggukjang produced at different parts in Korea. They all inhibited Listeria monocytogenes ATCC 19111 and nine inhibited Bacillus cereus ATCC 14579. Four isolates (W42, H27, SKE 12, and K21) showing strong inhibiting activities were identified as B. subtilis. B. subtilis W42 was the most inhibiting strain. The antimicrobial activity of culture supernatant from B. subtilis W42 was destroyed completely by proteinase K treatment, indicating that a bacteriocin was the responsible agent. The bacteriocin, Bac W42, was most stable at pH 7 and stable between pH 3-6 and 8-9. Bac W42 was stable up to $80^{\circ}C$. BHI (brain heart infusion) and TSB (tryptic soy broth) were the best media for the activity (320 AU/ml) followed by LB (160 AU/ml). Bac W42 was partially purified by column chromatographies. The specific activity was increased from 1,151.2 AU/ml to 9,043.5 AU/ml and the final yield was 26.3%. Bac W42 was 5.4 kDa in size as determined by SDS-PAGE. Bac W42 showed bactericidal activity against L. monocytogenes ATCC 19111.

• Research in Plant Disease
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• v.18 no.3
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• pp.201-209
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• 2012

The multi-function of 18 Bacillus subtilis isolates collected from agricultural extension centers of local government and National Academy of Agricultural Science was investigated by measuring their antifungal activities against five plant pathogens, such as Rhizoctonia solani, Colletotrichum acutatum, Fusarium oxysporum, Magnaporthe oryzae and Phytophthora capsici, phosphorus solubilization ability, production of indole acetic acid (IAA) and siderophore, and nitrogen fixation. The B. subtilis isolates showed antifungal activity against several plant pathogens and nitrogen fixation activity, and produced siderophore and IAA. They could control pepper powdery mildew (Leveillula taurica), but there was no difference in control efficacy among the B. subtilis isolates. In fields, the control efficacy of B. subtilis R2-1 ($10^8$ cells/ml) was compared with two microbial fungicides, Q-pect and Topsid. In 2009, the control efficacy of B. subtilis R2-1 (37.7%) was lower than that of Topsid (47.6%), but higher than that of Q-pect (25.7%). In 2010, the control efficacy of B. subtilis R2-1 (83.3%) was higher than that of Topsid (67.9%). In order to elucidate mode of action of B. subtilis R2-1 for controlling pepper powdery mildew, spore germination rates of pepper powdery mildew pathogen collected on treated leaves was investigated when suspensions of B. subtilis R2-1 and two microbial fungicides (Q-pect and Topsid) were foliar-sprayed. They highly suppressed spore germination of the pathogen with inhibition values of 84.2% for B. subtilis R2-1, 97.9% for Q-pect and 94.7% for Topsid. Further study on the mass-culturing method and formulation is needed for development of a microbial fungicide.

• Korean Journal of Organic Agriculture
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• v.6 no.1
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• pp.85-98
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• 1997

Cultural conditions for mass production of the antagonistic bacteria, Bacillus subtills CAP141 against pathogens causing major airborne diseases to apple trees, effects of temperature, pH, carbon and nitrogen source in the culture broth were investigated. The bacterial growth was most vigorous when the temperature and pH of the culture broth was 30~35$^{\circ}$C and 7, respectively. As for carbon source, dextrose was best followed in order by dextrose(monosaccharide) > sucrose(disaccharide) $\geq$ saccharose (market disaccharides) > starch (polysaccharieds). Among different sugars, bacterial growth was favored by in the order of brown, black and white supars, indicating that the bacterial growth might be promoted by the minor elements presented as impurities in the less purified sugars. As for nitrogen source, organic forms were better to bacterial growth than inorganic forms, that is polypeptone was best followed in order by soy sauce, soybeen milk and inorganic nitrogens. Differences in bacterial growth among different forms of inorganic nitrogen were negligible.

• KSBB Journal
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• v.15 no.4
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• pp.370-374
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• 2000

Some bacterial strains isolated from activated sludges and media and type cultures were cultivated in a pharmaceutical wastewater and a paperboard wastewater and added during batch treatment of those wastewaters in order for these strains to increase the treatment efficiency. Bacillus sp(PC-3) isolated from the charcoal media of the pharmaceutical wastewater plant grew remarkably over there strains in that wastewater and the viable cell count after 24hr cultivation was $1.1{\times}10^6m/L$. Bacillus subtills KCTC 1028 a type strain grew best in the paperboard wastewater and the viable cell count after 24hr cultivation was $1.1{\times}10^7m/L$. Addition of PC-3 in a batch treatment of the pharmaceutical wastewater increased COD removal by 18% after 8 day. And addition of Bacillus subtills KCTC 1028 in a batch treatment of the paperboard wastewater increased COD removal by 14% only after 24hy Bacillus subtills DCTC 1028 was though to be able to be produced economically using alcohol distillery wastewaters from starch material.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.59-62
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• 2008

The peptide methionine sulfoxide reductases (Msrs) are enzymes that catalyze the reduction of methionine sulfoxide back to methionine. Because of two enantiomers of methionine sulfoxide (S and R forms), this reduction reaction is carried out by two structurally unrelated classes of enzymes, MsrA (E.C. 1.8.4.11) and MsrB (E.C. 1.8.4.12). Whereas MsrA has been well characterized structurally and functionally, little information on MsrB is available. The recombinant MsrB from Bacillus subtilis has been purified and crystallized by the hanging-drop vapor-diffusion method, and the functional and structural features of MsrB have been elucidated. The crystals belong to the trigonal space group P3, with unit-cell parameters a=b=136.096, $c=61.918{\AA}$, and diffracted to $2.5{\AA}$ resolution using a synchrotron-radiation source at Pohang Light Source. The asymmetric unit contains six subunits of MsrB with a crystal volume per protein mass $(V_M)\;of\;3.37{\AA}^3\;Da^{-1}$ and a solvent content of 63.5%.