• Food Science and Preservation
• /
• v.17 no.3
• /
• pp.410-417
• /
• 2010

Several extracellular protease-producing bacteria were isolated from Chungkookjang, a traditional Korean food of fermented soybeans, on skim milk agar plates. Among these bacteria, strain D14 exhibited the highest production (15.2 U/mL) and specific activity (40.0 U/mg protein) of extracellular protease activity as assessed on growth in a protease induction medium composed of 1% (w/v) soluble starch, 1.5% (w/v) skim milk, 0.5% (w/v) yeast extract, and 2% (w/v) NaCl. The bacterium was identified as Bacillus subtilis based on morphological and physiological characteristics and 16S rDNA sequence. A BLAST search of 16S rDNA sequences revealed that the isolate was most closely related to Bacillus subtilis subsp. subtilis strain NCIB 3610. The 16S rDNA sequence homology was 99.9%. Our isolate produced the highest level of protease when grown in a protease induction medium containing 1% (w/v) sorbitol and 0.5% (w/v) yeast extract. Fructose and glucose reduced enzyme production to 12.7% and 35.9%, respectively, of the level seen when the strain was grown in medium containing soluble starch. Soytone also reduced enzyme production to 61.4% of the level noted when the strain was grown in medium containing yeast extract.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.5
• /
• pp.585-591
• /
• 2014

To evaluate the potential utility of pretreatment of raw biomass with a complex microbial system, we investigated the degradation of rice straw by BMC-9, a lignocellulose decomposition strain obtained from a biogas slurry compost environment. The degradation characteristics and corresponding changes in the bacterial community were assessed. The results showed that rapid degradation occurred from day 0 to day 9, with a peak total biomass bacterium concentration of $3.3{\times}10^8$ copies/ml on day 1. The pH of the fermentation broth declined initially and then increased, and the mass of rice straw decreased steadily. The highest concentrations of volatile fatty acid contents (0.291 mg/l lactic acid, 0.31 mg/l formic acid, 1.93 mg/l acetic acid, and 0.73 mg/l propionic acid) as well as the highest xylanse activity (1.79 U/ml) and carboxymethyl cellulase activity (0.37 U/ml) occurred on day 9. The greatest diversity among the microbial community also occurred on day 9, with the presence of bacteria belonging to Clostridium sp., Bacillus sp., and Geobacillus sp. Together, our results indicate that BMC-9 has a strong ability to rapidly degrade the lignocelluloses of rice straw under relatively inexpensive conditions, and the optimum fermentation time is 9 days.

• Journal of Food Hygiene and Safety
• /
• v.15 no.1
• /
• pp.51-54
• /
• 2000

In the 5'-flanking region of the D-AAT, AspAT and AlaDH gene, I found three or two pairs of sequences(designated as Pl, P2, P3) which show significant similarity to the E.coli consensus sequences of -35 and -10 for promoters. The spacing between -35 and -10 is 16 to 18bp in all the three putative promoters Pl, P2 and P3 which is in good agreement with the preferred spacer length in E.coli and in B.subtilis. Therefore, the putative promoters may also function to increase the efficiency of transcriptional initiation. The most stable, double-helical“Shine-Dalgarno”pairing is formed with a free energy change(ΔG) of -13.0 kcal/mol, -9.6 kcal/mol, -15.8 kcal/mol.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.28 no.3
• /
• pp.347-353
• /
• 1995

[ $F_o-value$ ] of canned smoked-oyster in cottonseed oil (SOCO) were measured using a microcomputer aided $F_o-value$ measuring system, and the microbiological safety of the canned SOCO was evaluated to optimize the energy consumption. Most of the microorganisms in raw oyster were saprophytes. No microorganisms were detected from the canned SOCO which was pretreated by conventional procedure and sterilized at $110^{\circ}C$ with $F_o-value$ of 5.92min and over. The most heat resistant microflora isolated from the raw oyster was Bacillus sp.. D-value at $121.1^{\circ}C$ and z-value of spores of Bacillus sp. in the SOCO homogenate were 4.10min and $10,91^{\circ}C$, respectively. After 120 days storage at $50^{\circ}C$, no growth of microorganisms was recognized from the canned SOCO with $F_o-value$of 5.92min.

• Food Science and Preservation
• /
• v.21 no.3
• /
• pp.442-450
• /
• 2014

In this study, we isolated a cellulase-producing bacterium isolated from traditional Korean fermented soybean paste and investigated the effect of culture conditions on the production of cellulase. This bacterium, which was identified as Bacillus subtilis 4-1 through 16S rRNA gene sequence analysis, showed the highest cellulase activity when the cells were grown at $45^{\circ}C$ for 24 hours in the CMC medium supplemented with 1.0% of soluble starch and 0.1% yeast extract. The initial optimum pH of the medium was observed in the range of 5.0~9.0. The optimal pH and temperature for the production of cellulase from B. subtilis 4-1 were pH 9.0 and $60^{\circ}C$ respectively. In addition, the enzyme showed significant activity in the temperature range of $20{\sim}90^{\circ}C$, which indicates that B. subtilis 4-1 cellulase is an alkaline-resistance and thermo-stable enzyme. This enzyme showed higher activity with CMC as the substrate for endo-type cellulase than avicel or pNPG as the exo-type substrates for exo-type cellulase and ${\beta}$-glucosidase. These results suggest that the cellulase produced from B. subtilis 4-1 is a complex enzyme rather than a mono-enzyme.

• Journal of Animal Science and Technology
• /
• v.49 no.4
• /
• pp.491-500
• /
• 2007

Two experiments were used to determine the effects of environment-friendly diets on growth performance, fecal excretion, nitrogen excretion and emission gases in manure for growing pigs. In experiment 1, ninety six crossed pigs(Landrace×Yorkshire×Duroc) were allocated into four treatments. Treatments were AME(adequate ME diet, 3,265 kcal/kg), LME(lower ME diet, 3,100 kcal/kg), LME 0.05(lower ME diet+α- galactosidase & β-mannanase 0.05%) and LME 0.10(lower ME diet+α-galactosidase & β-mannanase 0.10%). Pigs fed AME diet had lower ADFI(Average Daily Feed Intake) than pigs fed other diets(p<0.05). DM(Dry Matter) digestibility in pigs fed AME and LME 0.10 diets had greater than pigs fed LME diet(p<0.05). Energy digestibility is higher in pigs fed AME and LME 0.10 diets than other treatments(p<0.05). In experiment 2, twenty four crossbred pigs(33.71 kg average BW) were used in a 14-d metabolism experiment. The pigs were housed in individual cages equipped with plastic bed flooring. Treatments were CP(Crude protein) 18% without Bacillus sp., CP 18% diet+Bacillus sp. 0.05%, CP 14% without Bacillus sp. and CP 14% diet+Bacillus sp. 0.05%. Nitrogen intake was higher for CP 18% diets than CP 14% diets(p<0.05). DM, N(Nitrogen) and energy digestibility were affected by probiotics(p<0.05). With the high CP in diets, Energy and N digestibility, urine N percent, urine N excretion and total N excretion were increased significantly compared to low CP in diets(p<0.05). Among the treatments, DM and N digestibilities, feces N excretion, N absorption were decreased significantly(p<0.05), however, feces excretion, feces N, urine N percent, urine N excretion and total N excretion were increased significantly(p<0.05) when pigs fed without probiotics diets compare to pigs fed with probiotics diets. DM and N digestibility, feces excretion, feces N excretion, urine N percent, urine N excretion, total N excretion, N absorption and N adsorption ratio were CP×probiotic interactions in p<0.05. Ammonia(p<0.01) and H2S(p<0.05) in manure were lower in CP 14% diets than CP 18% diets. Also, ammonia and H2S in manure were CP×probiotic interactions in p<0.05. In conclusion, low energy and reduction of CP dietary added enzyme and probiotics improved nutrient digestibility and reduced odors emission in manure for growing pigs.

• Korean Journal of Microbiology
• /
• v.45 no.4
• /
• pp.377-384
• /
• 2009

Among 27 Bacillus sp. isolated from Meju, a traditional Korean soybean fermented food, a strain MJ-226 was selected due to its strong fibrinolytic activity, and it was identified to be Bacillus subtilis MJ-226 according to morphological and biochemical characterization and sugar utilization. The fibrinolytic enzyme of B. subtilis MJ-226 was maximally produced by cultivating in the Tryptic Soy Broth (TSB) for 24~26 h at $37^{\circ}C$, and the enzymes activity was promoted with adding glucose, fructose, peptone or yeast extract to TSB. The fibrinolytic enzyme was stable at the range of pH from 6.0 to 8.0, and between 35 and $40^{\circ}C$. Also, when the crude enzyme was exposed to various metal ions and chemical inhibitors for 12 h, the enzyme stability was maintained by $MnSO_4$, $CaCl_2$, KCl, and NaCl. However, the stability was destroyed by treatment with $CuSO_4$, $MgSO_4$, $ZnSO_4$, $FeSO_4$, and $BaCl_2$, and the enzyme was unstable in the presence of chemical inhibitors such as iodoacetic acid, leupeptin, phenylmethanesulphonyl fluoride (PMSF), sodium dodecyl sulfate (SDS), thiourea, trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA) and ethylenediaminetetraacetic acid (EDTA).

• Environmental Engineering Research
• /
• v.10 no.1
• /
• pp.1-6
• /
• 2005

Hexavalent chromium in aqueous solutions was successfully removed via sorption and reduction in the presence of waste leaves. Cr(VI) removal followed a first-order reaction, and removal rates were proportional to the amount of waste leaves used in the tests. Most of Cr(VI) were removed via sorption in early stages of the tests, but the reduction reaction played a significant role in Cr(VI) removal later. Solution pHs were continuously decreased due to the microbial activity, which was induced from the microorganisms attached on waste leaves. The decreased solution pHs further enhanced the sorption and reduction of Cr(VI). To characterize the microorganisms found in the tests, a denaturing gradient gel electrophoresis (DGGE) method was used. The majority of microorganisms were composed of Bacillus sp. which can reduce Cr(VI). Thus, waste leaves can be effective reactive media for the treatment of Cr(VI) in the subsurface.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.4
• /
• pp.527-537
• /
• 2019

Three types of meongge (Halocynthia roretzi) jeotgal (MJ) were prepared with 3 different types of salts (12%, w/v): purified salt (PS), solar salt aged for 3 years (SS), and bamboo salt that had been recrystalized 3 times (BS). One set of MJ was fermented with starters, Bacillus subtilis JS2 and Tetragenococcus halophilus BS1-37 (each 6 log CFU/g), and another set without starters for 42 days at $10^{\circ}C$. The LAB count of the SSMJ (non-starter) was highest at day 28 (2.30 log CFU/g). The pH of the PSMJ and SSMJ was 5.72-5.77 at day 0, and 5.40-5.50 at day 42. BSMJ showed higher pH and lower titratable acidities than other samples. Amino-type nitrogen (ANN) increased continuously, and SSMJ showed higher values than other samples from day 14. Bacterial species of non-starter MJ were examined by culture independent method. Clone libraries of 16S rRNA genes were constructed in Escherichia coli from total DNA from non-starter MJ samples at day 0, 14, and 28. Thirty clones per each sample were randomly selected and DNA sequences were analyzed. Variovorax sp., uncultured bacterium, and Acidovorax sp. were the most dominant group at day 0, 14, and 28, respectively. Lactobacillus sakei and Streptococcus sp. were the next dominant group in SSMJ at day 28. A Streptococcus sp. was detected from PSMJ at day 28. Sensory evaluation for MJ samples at day 28 showed that SSMJ got higher overall acceptability scores. These results showed that solar salt can cause desirable changes in the microbial community of fermented foods, thereby positively affecting their overall quality.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.3
• /
• pp.347-356
• /
• 2019

Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods. B. velezensis BS2 grows well at 15% NaCl and at $10^{\circ}C$. When B. velezensis BS2 was cultivated in TSB broth for 96 h at $37^{\circ}C$, the culture showed the highest fibrinolytic activity ($131.15mU/{\mu}l$) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and $37^{\circ}C$, respectively. $K_m$ and $V_{max}$ were 0.15 mM and $39.68{\mu}M/l/min$, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong ${\alpha}$-fibrinogenase and moderate ${\beta}$-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at $10^{\circ}C$, B. velezensis BS2 can be used as a starter for jeotgal.