• Title/Summary/Keyword: B. subtilis p-4

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Expression and Biochemical Characterization of Cold-Adapted Lipases from Antarctic Bacillus pumilus Strains

  • Litantra, Ribka;Lobionda, Stefani;Yim, Joung Han;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1221-1228
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    • 2013
  • Two lipase genes (bpl1 and bpl3) from Antarctic Bacillus pumilus strains were expressed in Bacillus subtilis. Both recombinant lipases BPL1 and BPL2 were secreted to the culture medium and their activities reached 3.5 U/ml and 5.0 U/ml, respectively. Their molecular masses apparent using SDS-PAGE were 23 kDa for BPL1 and 19 kDa for BPL3. Both lipases were purified to homogeneity using ammonium sulfate precipitation and HiTrap SP FF column and Superose 12 column chromatographies. The final specific activities were estimated to be 328 U/mg for BPL1 and 310 U/mg for BPL3. Both lipases displayed an optimum temperature of $35^{\circ}C$, similar to other mesophilic enzymes. However, they maintained as much as 70% and 80% of the maximum activities at $10^{\circ}C$. Accordingly, their calculated activation energy at a temperature range of $10-35^{\circ}C$ was 5.32 kcal/mol for BPL1 and 4.26 kcal/mol for BPL3, typical of cold-adapted enzymes. The optimum pH of BPL1 and BPL3 was 8.5 and 8.0, respectively, and they were quite stable at pH 7.0-11.0, showing their strong alkaline tolerance. Both lipases had a preference toward medium chain length ($C_6-C_{10}$) fatty acid substrates. These results indicate the potential for the two Antarctic B. pumilus lipases as catalysts in bioorganic synthesis, food, and detergent industries.

Comparison of Antioxidant and Antimicrobial Activities of Bracken (Pteridium aquilinum Kuhn) according to Cooking Methods (조리방법에 따른 고사리의 항산화활성 및 항균활성 비교)

  • Park, Cho-Hee;Kim, Kyoung-Hee;Yook, Hong-Sun
    • The Korean Journal of Food And Nutrition
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    • v.27 no.3
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    • pp.348-357
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    • 2014
  • This study was carried out to evaluate the yield of extract, antioxidant compounds (total phenolic and total flavonoid), antioxidant (DPPH assay, ABTS assay and reducing power), and antimicrobial activities of bracken (Pteridium aquilinum Kuhn), according to cooking methods (non-blanched, blanched and seasoned). The yield of seasoned bracken extracts showed a high value of (4.59%) followed by non-blanched bracken and blanched bracken with 2.69% and 0.30%, respectively. In the total polyphenol and flavonoid contents, seasoned bracken extracts showed higher antioxidant compounds ($96.11{\pm}0.34mg\;GAE$/100 g RW, $20.90{\pm}0.mg\;CE$/100 g RW) than non-blanched and blanched. The total antioxidant activities (DPPH assay, ABTS assay and reducing power) were shown to be in the order of seasoned bracken > non-blanched bracken > blanched bracken. In the antimicrobial activities, non-blanched bracken extracts showed antimicrobial activity against B. cereus, B. subtilis, E. cloacae, E. coli, S. enterica, and P. aeruginosa except for S. aureus. The non-blanched bracken extracts (5 and 10 mg/disc) especially showed strong antimicrobial activity against P. aeruginosa ($10.00{\pm}0.71$ and $10.25{\pm}0.35mm$). The inhibition zone diameter from the extracts of blanched bracken and seasoned bracken was not detected. Many seasonings added in the process of cooking can increase the antioxidant capacities. The overall results of this study demonstrate that the cooked bracken with seasoning would be the most efficient way of ingesting antioxidant compounds.

Enhancement of the Thermostability of a Fibrinolytic Enzyme from Bacillus amyloliquefaciens CH51 (Bacillus amyloliquefaciens CH51이 생산하는 혈전용해효소의 열안정성 개선)

  • Kim, Jieun;Choi, Kyoung-Hwa;Kim, Jeong Hwan;Song, Young-Sun;Cha, Jaeho
    • Journal of Life Science
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    • v.23 no.1
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    • pp.15-23
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    • 2013
  • AprE51 from Bacillus amyloliquefaciens CH51 is a 27 kDa subtilisin-like protease with fibrinolytic activity. AprE51-6 showing increased catalytic activity was produced previously. To enhance the thermostability of AprE51-6, 2 residues, Gly-166 and Asn-218 based on B. subtilis subtilisin E were mutated by site-directed mutagenesis. The results of the mutational analysis showed that substitution of arginine for Gly-166 (AprE51-7) increased the fibrinolytic activity 1.8-fold. An N218S mutant (AprE51-8) also increased the fibrinolytic activity up to 4.5-fold in a fibrin plate assay. Purified AprE51-7 and AprE51-8 mutants had a 1.9- and a 2.5-fold higher $k_{cat}$, respectively, and a 2.1-1.9-fold lower $K_m$, respectively. This resulted in a 3.8- and a 4.7-fold increase in catalytic efficiency ($k_{cat}/K_m$), respectively, relative to that of wild-type AprE51. AprE51-8 had a broader pH range than AprE51-6 and nattokinase, especially at an alkaline pH value. In addition, AprE51-8 showed higher thermostability than AprE51-6 at $60^{\circ}C$. The half-lives of AprE51-7 and AprE51-8 at $50^{\circ}C$ were 21.5 and 27.3 min, respectively, which are 2.0 and 2.6 times longer, respectively, than that of the wild-type AprE51.

Biochemical and Molecular Identification of Antibacterial Lactic Acid Bacteria Isolated from Kimchi (김치에서 항균활성 유산균의 분리 및 동정)

  • Kim, Soo-Young;Kim, Jong-Doo;Son, Ji-Soo;Lee, Si-Kyung;Park, Kab-Joo;Park, Myeong-Soo
    • Korean Journal of Food Science and Technology
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    • v.43 no.4
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    • pp.446-452
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    • 2011
  • Total 480 lactic acid-producing bacteria were isolated from five kinds of kimchi, and their antibacterial activity was tested against Salmonella enterica serovar Typhimurium, Bacillus subtilis, and Pseudomonas aeruginosa using an agar diffusion assay. Among them, 340 isolates showed a halo on MRS agar against one or more indicator strains, which were identified using multiplex PCR, an API 50CHL kit, and a 16S rDNA sequence analysis. As a result, 169 Lactobacillus plantarum, 20 Lactobacillus fermentum, two Lactobacillus paracasei ssp. paracasei, two Lactobacillus sp., and 15 Pediococcus sp. were identified. This may be the first report on the isolation of antibacterial Lactobacillus fermentum from kimchi.

Green Synthesis of Copper Nano-Drug and Its Dental Application upon Periodontal Disease-Causing Microorganisms

  • El-Rab, Sanaa M.F. Gad;Basha, Sakeenabi;Ashour, Amal A.;Enan, Enas Tawfik;Alyamani, Amal Ahmed;Felemban, Nayef H.
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1656-1666
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    • 2021
  • Dental pathogens lead to chronic diseases like periodontitis, which causes loss of teeth. Here, we examined the plausible antibacterial efficacy of copper nanoparticles (CuNPs) synthesized using Cupressus macrocarpa extract (CME) against periodontitis-causing bacteria. The antimicrobial properties of CME-CuNPs were then assessed against oral microbes (M. luteus. B. subtilis, P. aerioginosa) that cause periodontal disease and were identified using morphological/ biochemical analysis, and 16S-rRNA techniques. The CME-CuNPs were characterized, and accordingly, the peak found at 577 nm using UV-Vis spectrometer showed the formation of stable CME-CuNPs. Also, the results revealed the formation of spherical and oblong monodispersed CME-CuNPs with sizes ranged from 11.3 to 22.4 nm. The FTIR analysis suggested that the CME contains reducing agents that consequently had a role in Cu reduction and CME-CuNP formation. Furthermore, the CME-CuNPs exhibited potent antimicrobial efficacy against different isolates which was superior to the reported values in literature. The antibacterial efficacy of CME-CuNPs on oral bacteria was compared to the synergistic solution of clindamycin with CME-CuNPs. The solution exhibited a superior capacity to prevent bacterial growth. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and fractional inhibitory concentration (FIC) of CME-CuNPs with clindamycin recorded against the selected periodontal disease-causing microorganisms were observed between the range of 2.6-3.6 ㎍/ml, 4-5 ㎍/ml and 0.312-0.5, respectively. Finally, the synergistic antimicrobial efficacy exhibited by CME-CuNPs with clindamycin against the tested strains could be useful for the future development of more effective treatments to control dental diseases.

Antimicrobial, Antioxidant and Anticoagulation Activities of Korean Radish (Raphanus sativus L.) Leaves (무청의 항균, 항산화 및 항혈전 활성)

  • Lee, Ye-Seul;Kwon, Kyung-Jin;Kim, Mi-Sun;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.228-235
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    • 2013
  • Radish (Raphanus sativus) is a common cruciferous vegetable, and its aerial parts, called Mu-chung in Korean, have plentiful nutritional components such as vitamins, minerals and dietary fibers. Mu-chung has been used as a kimchi, a traditional Korean fermented dish, and dried Mu-chung is an important component of soups commonly consumed during winter in Korea. Since the advent of the mass production of radish in Korea, with the segregation of farm areas and towns and changing diets, Mu-chung has mostly been discarded instead of utilized. In addition, studies concerning the efficient utilization and useful bioactivities of Mu-chung are still lacking worldwide. In this study, we prepared the ethanol extract of Mu-chung and its subsequent solvent fractions. Antimicrobial, antioxidation, and anticoagulation activities were then evaluated in the hopes of developing a functional biomaterial from Korean radishes' aerial parts. The ethanol extraction yield for hot-air dried Mu-chung was 5.6%, and the fraction yields of n-hexane (H), ethylacetate (EA), butanol (B) and water residue were 25.3, 3.6, 19.4, and 51.7%, respectively. Analysis of total polyphenol and total flavonoid contents showed that the EA fraction had the highest content (97.57 and 152.91 mg/g) amongst the fractions. In antimicrobial activity assays, the H and EA fractions were effective against gram positive bacteria (Staphylococcus aureus, Listeria monocytogenes, and Bacillus subtilis), but not effective against gram negative bacteria (Escherichia coli and Pseudomonas aeruginosa). The B fraction also exhibited moderate antibacterial activity, suggesting that the extract of Mu-chung has various antibacterial components. In antioxidation activity assays, the EA fraction showed strong DPPH, ABTS and nitrite scavenging activities ($69-222{\mu}g/ml$ of $IC_{50}$), including reducing power. In anticoagulation activity assays, the EA fraction demonstrated strong inhibition activity against human thrombin and prothrombin. Prominent anticoagulation activity was found in aPTT assays; the aPTT of the EA fraction was extended 15-fold compared than that of the solvent control. Our results suggest that Mu-chung is an attractive nutritional food material possessing useful bioactivities, and the EA fraction of Mu-chung could be developed as a functional food ingredient.

Esterase Production and Culture Characteristics of Bacteria Isolated from Acid Hydrolysed Soybean Protein (산분해 대두 단백질로부터 분리된 Esterase 생성균의 생육 및 효소생성 특성)

  • Oh, Nam-Soon
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.484-489
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    • 1997
  • The characteristics of growth and esterase activity of bacterial strains isolated from acid hydrolysed soybean protein were examined. All the isolated strains having decomposition activity of p-hydroxybenzoic acid butyl ester and esterase producing activity were identified as Bacillus sp. by morphological and biochemical methods. The specific growth rates, esterase activities and p-hydroxybenzoic acid butyl ester decomposition activities of isolated strains were $0.844{\sim}1.213\;h^{-1}$, $21{\sim}222\;mU/ml$ and $5.4{\sim}8.1\;mU/ml$, respectively. In the fermentation of Bacillus sp. KB8 strain which had the highest esterase producing activity, growth, extracellular excretion and intracellular synthesis of esterase were inhibited by adding NaCl in the culture broth. Esterase producing activity gradually increased after late exponential growth phase, until maximum value of 420 mU/ml reached after 64 hours culture period. Esterase of Bacillus sp. KB8 strain was stable up to $50^{\circ}C$ for 30 minutes, but was inactivated by heating for 30 minutes at $70^{\circ}C$. The enzyme activity exponentially decreased during the incubation time at the temperatures of $60^{\circ}C$ and $65^{\circ}C$.

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The Studies on the Removal of Harmful Microorganisms in Water by Using Antimicrobial Copper (친환경 항균성 구리의 수질 내 생물학적 오염원 제거 가능성 연구)

  • Ji, Keunho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.9
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    • pp.384-390
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    • 2018
  • Air and soil are being contaminated by the environmental pollution as a result of climate change and urbanization, resulting in water pollution reaching serious levels. In this studies, we investigated the use of antimicrobial copper for the removal of biological pollutants from water system. Specifically, we tested its effects against E. coli, B. subtilis, S. aureus, K. pneumoniae and P. aeruginosa. Made a sphere shape having a diameter of 2cm using a strip-shaped copper wire of 0.5g, 1g and 2g. And then, to confirm the antimicrobial activities, each copper ball was equipped in the broth which inoculated each pathogens. The results showed that bacterial growth of the five test bacteria was inhibited by more than 99% after reaction with a 0.5 g copper ball for at least 20 minutes. Based on the these results, if perform the further experiment such cytotoxicity, it is expected that will be enough to be used as a filter for water quality purification. The developed technique is expected to be widely applied in various industries.

Antimicrobial Activity of Aqueous Ethanol Extracts of Perilla frutescens var. acuta Leaf (차조기의 에탄올과 물 혼합 추출물의 항균활성)

  • Kim, Mi-Hyang;Lee, Nan-Hee;Lee, Myung-Hee;Kwon, Dae-Jun;Choi, Ung-Kyu
    • Journal of the Korean Society of Food Culture
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    • v.22 no.2
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    • pp.266-273
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    • 2007
  • This study was conducted to examine the antimicrobial activity of Perilla frutescens var. acuta leaf fractions extracted with a mixture of ethanol and water. The Ca and Mg contents of the leaf were 595.75 mg% and 467.0 mg%, respectively, and they were the highest among all of the test minerals. The extract yield increased w e content of water in e extraction solvent. Antimicrobial activity against Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa was found in the 50,70 and 95% ethanol extracts. Of the various fractions extracted from the mixture of ethanol and water, the ethyl acetate fraction showed antibacterial activity against all microorganisms tested in this experiment, and the antibacterial activity of ethyl acetate fraction from the water extract was the strongest. The phenol and flavonoid content in the ethyl acetate fraction showed no correlation with the concentration of ethanol in the extract solvent; however, their contents were higher in the 30% ethanol and water extraction which the antimicrobial activity of the extract was the strongest.

Antioxidative and Antimicrobial Effects of Water Extracts with Castanea crenata Leaf Tea (밤잎차(茶) 물추출물의 항산화 및 항미생물 효과)

  • Choi, Ok-Beom;Yoo, Gyurng-Soo;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.1128-1131
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    • 1999
  • Antioxidative and antimicrobial effects of the extracts of Castanea crenata leaf tea, steamed tea and semi-fermented tea were investigated. Antioxidative effects were by measuring the 1,1-diphenyl-2-picrylhydrazyl(DPPH)-radical scavening activity of the extracts. The effects were stronger than that of synthetic antioxidant such as BHT at the same concentration. The $SC_{50}$ value (50% radical scavening effects of $1{\times}10^{-4}$ M DPPH) of steamed tea, semi-fermented tea and BHT were 53.3 ${\mu}g/mL$, 49.9 ${\mu}g/mL$ and 101.0 ${\mu}g/mL$, respectively. The antimicrobial effects of methanol extracts (8 mg, 20 mg) of 0.2 g and 0.5 g. eq. of Castanea crenata leaf tea were stronger than those of 0.65 mg of benzoic acid, against gram(+) bacteria such as S. aureus, S. epidermidis, M. luteus, L. mesenteroides and B. subtilis and gram(-) bacteria such as E. coli, S. typhimurium, and P. aeruginosa.

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