• Title/Summary/Keyword: B.subtilis

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A Two-Strain Mixture of Rhizobacteria Elicits Induction of Systemic Resistance Against Pseudomonas syringae and Cucumber Mosaic Virus Coupled to Promotion of Plant Growth on Arabidopsis thaliana

  • Ryu Choong-Min;Murphy John F.;Reddy M.S.;Kloepper Joseph W.
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.280-286
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    • 2007
  • We evaluated a commercial biopreparation of plant growth-promoting rhizobacteria (PGPR) strains Bacillus subtilis GB03 and B. amyloliquefaciens IN937a formulated with the carrier chitosan (Bio Yield) for its capacity to elicit growth promotion and induced systemic resistance against infection by Cucumber Mosaic Virus (CMV) and Pseudomonas syringae pv. tomato DC3000 in Arabidopsis thaliana. The biopreparation promoted plant growth of Arabidopsis hormonal mutants, which included auxin, gibberellic acid, ethylene, jasmonate, salicylic acid, and brassinosteroid insensitive lines as well as each wild-type. The biopreparation protected plants against CMV based on disease severity in wild-type plants. However, virus titre was not lower in control plants and those treated with biopreparation, suggesting that the biopreparation induced tolerance rather than resistance against CMV. Interestingly, the biopreparation induced resistance against CMV in NahG plants, as evidenced by both reduced disease severity and virus titer. The biopreparation also elicited induced resistance against P. syringae pv. tomato in the wild-type but not in NahG transgenic plants, which degrade endogenous salicylic acid, indicating the involvement of salicylic acid signaling. Our results indicate that some PGPR strains can elicit plant growth promotion by mechanisms that are different from known hormonal signaling pathways. In addition, the mechanism for elicitation of induced resistance by PGPR may be pathogen-dependent. Collectively, the two-Bacilli strain mixture can be utilized as a biological inoculant for both protection of plant against bacterial and viral pathogens and enhancement of plant growth.

Identification and Characterization of a New Alkaline Thermolysin-Like Protease, BtsTLP1, from Bacillus thuringiensis Serovar Sichuansis Strain MC28

  • Zhang, Zhenghong;Hao, Helong;Tang, Zhongmei;Zou, Zhengzheng;Zhang, Keya;Xie, Zhiyong;Babe, Lilia;Goedegebuur, Frits;Gu, Xiaogang
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1281-1290
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    • 2015
  • Thermolysin and its homologs are a group of metalloproteases that have been widely used in both therapeutic and biotechnological applications. We here report the identification and characterization of a novel thermolysin-like protease, BtsTLP1, from insect pathogen Bacillus thuringiensis serovar Sichuansis strain MC28. BtsTLP1 is extracellularly produced in Bacillus subtilis, and the active protein was purified via successive chromatographic steps. The mature form of BtsTLP1 has a molecule mass of 35.6 kDa as determined by mass spectrometry analyses. The biochemical characterization indicates that BtsTLP1 has an apparent Km value of 1.57 mg/ml for azocasein and is active between 20℃ and 80℃. Unlike other reported neutral gram-positive thermolysin homologs with optimal pH around 7, BtsTLP1 exhibits an alkaline pH optimum around 10. The activity of BtsTLP1 is strongly inhibited by EDTA and a group of specific divalent ions, with Zn2+ and Cu2+ showing particular effects in promoting the enzyme autolysis. Furthermore, our data also indicate that BtsTLP1 has potential in cleaning applications.

Improved Production of Long-Chain Fatty Acid in Escherichia coli by an Engineering Elongation Cycle During Fatty Acid Synthesis (FAS) Through Genetic Manipulation

  • Jeon, Eunyoung;Lee, Sunhee;Lee, Seunghan;Han, Sung Ok;Yoon, Yeo Joon;Lee, Jinwon
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.990-999
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    • 2012
  • The microbial biosynthesis of fatty acid of lipid metabolism, which can be used as precursors for the production of fuels of chemicals from renewable carbon sources, has attracted significant attention in recent years. The regulation of fatty acid biosynthesis pathways has been mainly studied in a model prokaryote, Escherichia coli. During the recent period, global regulation of fatty acid metabolic pathways has been demonstrated in another model prokaryote, Bacillus subtilis, as well as in Streptococcus pneumonia. The goal of this study was to increase the production of long-chain fatty acids by developing recombinant E. coli strains that were improved by an elongation cycle of fatty acid synthesis (FAS). The fabB, fabG, fabZ, and fabI genes, all homologous of E. coli, were induced to improve the enzymatic activities for the purpose of overexpressing components of the elongation cycle in the FAS pathway through metabolic engineering. The ${\beta}$-oxoacyl-ACP synthase enzyme catalyzed the addition of acyl-ACP to malonyl-ACP to generate ${\beta}$-oxoacyl-ACP. The enzyme encoded by the fabG gene converted ${\beta}$-oxoacyl-ACP to ${\beta}$-hydroxyacyl-ACP, the fabZ catalyzed the dehydration of ${\beta}$-3-hydroxyacyl-ACP to trans-2-acyl-ACP, and the fabI gene converted trans-2-acyl-ACP to acyl-ACP for long-chain fatty acids. In vivo productivity of total lipids and fatty acids was analyzed to confirm the changes and effects of the inserted genes in E. coli. As a result, lipid was increased 2.16-fold higher and hexadecanoic acid was produced 2.77-fold higher in E. coli JES1030, one of the developed recombinants through this study, than those from the wild-type E. coli.

Study on the Screening and Development of Antibiotics in the Mushrooms -The Screening of Bacterial Antibiotics in Basidiomycetes (I)- (버섯 중 항균물질의 검색 및 개발에 관한 연구 -그람양성균에 대한 항균물질의 검색 (1보)-)

  • Park, Sang-Shin;Lee, Kap-Duk;Min, Tae-Jin
    • The Korean Journal of Mycology
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    • v.23 no.1 s.72
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    • pp.28-36
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    • 1995
  • To investigate the antibacterial activities of 53 species of mushrooms for Gram-positive bacteria, bioactive substances from mushrooms were extracted with petroleum ether, 80% ethanol, and distilled water. The 14 extracts of 12 species of mushrooms, largely with 80% ethanol, showed antibacterial activities against 6 species of Gram-positive bacteria selectively. Especially, the extracts of Thylopilus PS-5, Pulveroboletus PS-7, and Russula PS-9 showed various antibacterial activities against several bacteria. Minimal inhibitory concentration (MIC) of the 80% ethanol extracts from Pulveroboletus PS-7 and Boletus PS-8 was $300\;{\mu}g/ml$ against Bacillus subtilis, and that of the 80% ethanol extracts from Trichaptium PS-3 was $500\;{\mu}g/ml$ against Micrococcus luteus. MIC of other extracts with antibacterial activities was above $1000\;{\mu}g/ml$.

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Antimicrobial, Antiperspirant and Deodorant Effect of Herbal Extract Mixtures Including Schisandra chinensis (오미자를 함유한 복합물의 항균, 발한억제 및 소취효과)

  • Ahn, Ji-Young;Kim, Jun-Ho;Kwon, Se-Uk;Kim, Dae-Ki;Lee, Kang-Soo;Lee, Young-Mi
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.1
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    • pp.53-58
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    • 2012
  • The aim of this study was to investigate the antimicrobial, antiperspirant and deodorant effect of herb mixtures including Schisandra chinensis(SC). This herb mixtures including Schisandra chinensis was made of Astragalus membranaceus(AM), Triticum aestivum(TA), Atractylodes japonica(AJ) and peppermint essential oil extracted from Mentha piperita(MP). We examined antimicrobial activity, antiperspirant activity and L-leucine dehydrogenase activity of the herbal extracts. We found that the mixture of SC and MP have the best antimicrobial effects on B. subtilis subsp. spizizenii and mixture of SC, AJ, AM and TA have best antiperspirant effect. Moreover, the combination of SC and AM showed the most inhibitory effect on L-leucine dehydrogenase activity. These results suggested that the herb mixtures Including Schisandra chinensis have good antimicrobial and antiperspirant effects.

Study on the Physiological Activities of Cleyera japonica Extract (비쭈기 나무(Cleyera japonica) 추출물의 생리활성에 대한 연구)

  • Ahn, JoungJwa;Hwang, Tae-Young;Kim, Hyun-Soo
    • Korean Journal of Plant Resources
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    • v.28 no.2
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    • pp.153-157
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    • 2015
  • In this study, we investigated the applicability of functional materials by examining a variety of physiological activities with the extract of Cleyera japonica leaf. Cleyera japonica extract showed a low cytotoxicity against murine melanoma B16F10 cells. In little or no cytotoxicity at concentrations, we showed that the treatment with Cleyera japonica extract resulted in a significant increase in the DPPH radical scavenging activity (IC50, 22.90 ㎎/L), similar to ascorbic acid (IC50, 18.65 ㎎/L) and anti-microbial activities against Bacillus subtilis, Escherichia coli, and Candida albicans. In particular, anti-microbial activities against Gram-positive bacteria was high. These results suggest that Cleyera japonica extract could be used as a natural preservative. Additionally, Cleyera japonica extract showed the inhibition of tyrosinase activity (IC50, 178.90 ㎎/L), similar to kojic acid (IC50, 89.13 ㎎/L) and decreased melanin content (IC50, 101.90 ㎎/L) higher than the control arbutin level (IC50, 100.65 ㎎/L), especially. Therefore, these results indicate that Cleyera japonica extract may be an effective material for functional cosmetics such as skin whitening materials.

Replacement of Dietary Vitamin C with Citrus and Fermented Citrus By-products in Korean Rockfish Sebastes schlegelii (조피볼락(Sebastes schlegelii) 사료 내 합성 비타민C 대체원으로써의 감귤부산물과 발효 감귤부산물의 첨가 효과)

  • Shin, Jaehyeong;Song, Jin-Woo;Kim, Min-Gi;Lim, Hyunwoon;Lim, Jongho;Lee, Kyeong-Jun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.5
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    • pp.617-623
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    • 2021
  • The study was conducted to investigate the dietary supplementation of citrus by-product (CBP) and CBP fermented with Bacillus subtilis (BS) or B. pumilus (BP) on growth performance, feed utilization, innate immune responses and disease resistance of Korean rockfish Sebastes schlegelii. Triplicate groups of juvenile fish (initial weight of 22.9 g) were fed one of five experimental diets which were formulated to replace a synthetic vitamin C (L-ascorbyl-2-polyphosphate, LAPP) with CBP or fermented CBP. A basal diet without LAPP and CBP was used as the control diet (Con). At the end of the 13 weeks feeding trial, growth performance, feed utilization and survival of fish fed the Con were significantly lower than those of fish fed the LAPP, CBP and fermented CBP diets. Fish fed the Con were significantly lower in lysozyme activity than fish fed LAPP, CBP and fermented CBP diets. In a challenge test against Edwardsiella tarda, cumulative mortality of fish fed LAPP, CBP and fermented CBP diets was significantly lower (20-30%) than that of the Con (60%). The findings in this study indicate that the CBP and/or fermented CBP can be a promising vitamin C source for LAPP replacement in diet for Korean rockfish.

Crystal structure of unphosphorylated Spo0F from Paenisporosarcina sp. TG-14, a psychrophilic bacterium isolated from an Antarctic glacier

  • Lee, Chang Woo;Park, Sun-Ha;Jeong, Chang Sook;Lee, Chang Sup;Hong, Jong Wook;Park, Hyun Ho;Park, Hyun;Park, HaJeung;Lee, Jun Hyuck
    • Biodesign
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    • v.6 no.4
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    • pp.84-91
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    • 2018
  • Spo0F is a response regulator that modulates sporulation, undergoes phosphorylation for phosphorelay signal transduction, and interacts with various regulatory proteins; however, the mechanisms through which phosphorylation induces structural changes and regulates interactions with binding partners remain unclear. Here, we determined the unphosphorylated crystal structure of Spo0F from the psychrophilic bacterium Paenisporosarcina sp. TG-14 (PaSpo0F) and established a phosphorylation-state structural model. We found that PaSpo0F underwent structural changes (Lys54 and Lys102) by phosphorylation and generated new interactions (Lys102/Gln10 and Lys54/Glu84) to stabilize the ${\beta}4/{\alpha}4$ and ${\beta}1/{\alpha}1$ loop structures, which are important target-protein binding sites. Analysis of Bacillus subtilis Spo0 variants revealed movement by BsSpo0F Thr82 and Tyr84 residues following interaction with BsSpo0B, providing insight into the movement of corresponding residues in PaSpo0F (Thr80 and Tyr82), with further analysis of BsSpo0F/BsRapH interaction revealing alterations in the ${\beta}4/{\alpha}4$ loop region. These results suggest that phosphorylation-induced structural rearrangement might be essential for PaSpo0F activation and expand the understanding of Spo0F-specific activation mechanisms during sporulation.

Antagonistic Activity of Bacteria Isolated from Apple in Different Fruit Development Stages against Blue Mold Caused by Penicillium expansum

  • Lopez-Gonzalez, Rocio Crystabel;Juarez-Campusano, Yara Suhan;Rodriguez-Chavez, Jose Luis;Delgado-Lamas, Guillermo;Medrano, Sofia Maria Arvizu;Martinez-Peniche, Ramon Alvar;Pacheco-Aguilar, Juan Ramiro
    • The Plant Pathology Journal
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    • v.37 no.1
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    • pp.24-35
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    • 2021
  • Blue mold caused by Penicillium expansum is one of the most significant postharvest diseases of apples. Some microorganisms associated with the surface of ripening apples possess the ability to inhibit the growth of P. expansum. However, the existing literature about their colonization in the stages before ripening is not explored in depth. This study aims to characterize the antagonistic capacity of bacterial populations from five fruit development stages of 'Royal Gala' apples. The results have shown that the density of the bacterial populations decreases throughout the ripening stages of fruit (from 1.0 × 105 to 1.1 × 101 cfu/㎠). A total of 25 bacterial morphotypes (corresponding to five genera identified by 16S RNA) were differentiated in which Bacillus stood out as a predominant genus. In the in vitro antagonism tests, 10 Bacillus strains (40%) inhibited the mycelial growth of P. expansum from 30.1% to 60.1%, while in fruit bioassays, the same strains reduced the fruit rot ranging from 12% to 66%. Moreover, the bacterial strains with antagonistic activity increased in the ripening fruit stage. B. subtilis subsp. spiziennii M24 obtained the highest antagonistic activity (66.9% of rot reduction). The matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis revealed that bacteria with antagonistic activity produce anti-fungal lipopeptides from iturin and fengycin families.

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.