• Title/Summary/Keyword: Bacillus Megaterium

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Assessment for $CO_2$ Biomineralization Characteristics and its Applicability for Solidified Sludge (이산화탄소 생광물화 특성 및 슬러지 고화물 적용성 평가)

  • Min, Dong-Hee;Ahn, Chang-Min;Han, Ji-Sun;Yoon, Soon-Uk;Jeon, Eun-Jeong;Won, Jong-Choul;Chun, Seung-Kyu;Kim, Chang-Gyun
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.11
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    • pp.812-820
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    • 2011
  • This study was conducted to characterize $CO_2$ biomineralization on several minerals (i.e., CaO, MgO, $SiO_2$) by bottle test in an aqueous solution and solidified sludge using different aerobic bacterial strains like Bacillus megaterium and Bacillus pasteurii by batch test. These bacteria promote the formation of microenvironments that facilitate the precipitation of mineral phases that were unsaturated in the bulk solution. For one type of mineral solely amended, the $CO_2$ was reduced at the highest of 4.0 mmol for MgO while it was not that much lower for CaO and $SiO_2$ showing 1.1 and 0.3 mmol $CO_2$2, respectively. For two types of minerals simultaneously amended, the $CO_2$ was reduced at the greater extent for both Ca + Mg and Mg + Si showing 2.7 and 2.3 mmol, respectively whereas it was less for Ca + Si at 1.8 mmol. For solidified sludge, the $CO_2$ reduction rate changed depending on the volume of solidified sludge placed in the medium and the input $CO_2$ concentration.. The reduction rate of $CO_2$ was increased with increasing the volume of solidified sludge. Results of XRD analysis indicate that $CaCO_3$ (Calcite) was dominantly formed among others (e.g., Aragonite, Dolomite). SEM analysis showed that the sample with Bacillus pasteurii, could more form minerals rather than control. As demonstrated in this study, $CO_2$ would be effectively sequestered in biomineralization process.

An Antifungal Antibiotic Purified from Bacillus megaterium KL39, a Biocontrol Agent of Red-Pepper Phytophthora-Blight Disease

  • JUNG HEE KYOUNG;KIM SANG-DAL
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1001-1010
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    • 2005
  • Bacillus megaterium KL39, an antibiotic-producing plant growth promoting rhizobacterium (PGPR), was selected from soil. The antifungal antibiotic, denoted KL39, was purified from culture filtrate by column chromatography using Dion HP-20, Silica gel, Sephadex LH-20, and prep-HPLC. Thin layer chromatography, employing the solvent system of ethanol:ammonia:water=8:1:1, showed the $R_{f}$. value of 0.32. The antibiotic KL39 showed a negative reaction with ninhydrin solution, positive with iodine vapor, and also positive with Ehrlich reagent. It was soluble in methanol, ethanol, butanol, and acetonitrile, but insoluble in chloroform, toluene, hexane, ethyl ether, or acetone. Its UV spectrum had the maximum absorption at 208 nm. Amino acid composition, FAB-mass, $^{1}H-NMR,\;^{13}C-NMR$, and atomic analyses showed that the antibiotic KL39 (MW=1,071) has a structure very similar to iturin E. The antibiotic KL39 has a broad antifungal spectrum against a variety of plant pathogenic fungi including Rhizoctonia solani, Pyricularia oryzae, Monilinia froeticola, Botrytis cinenea, Altenaria kikuchiana, Fusarium oxysporum, and F. solani. An MIC value of $10\;{\mu}g/ml$ was determined for Phytophthora capsici. Macromolecular incorporation studies with P. capsici using radioactive [$^{3}H-adenine$] as the precursor, indicated that the antibiotic KL39 strongly inhibits the DNA biosynthesis of the fungal cell. Microscopic observation of the antifungal action showed abnormal hyphal swelling of P. capsici. The purified antibiotic KL39 was very effective for the biocontrol of in vivo Phytophthora-blight disease of pepper.

Improvement of Biocontrol of Damping-off and Root Rot/Wilt of Faba Bean by Salicylic Acid and Hydrogen Peroxide

  • Abdel-Monaim, Montaser Fawzy
    • Mycobiology
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    • v.41 no.1
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    • pp.47-55
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    • 2013
  • Rhizoctonia solani, Fusarium solani, F. oxysporum, and Macrophomina phaseolina were found to be associated with root rott and wilt symptoms of faba bean plants collected from different fieldes in New Valley governorate, Egypt. All the obtained isolates were able to attack faba bean plants (cv. Giza 40) causing damping-off and root rot/wilt diseases. R. solani isolates 2 and 5, F. solani isolate 8, F. oxysporum isolate 12 and M. phaseolina isolate 14 were the more virulent ones in the pathogenicity tests. Biocontrol agents (Trichoderma viride and Bacillus megaterium) and chemical inducers (salicylic acid [SA] and hydrogen peroxide) individually or in combination were examined for biological control of damping-off and root rot/wilt and growth promoting of faba bean plants in vitro and in vivo. Both antagonistic biocontrol agents and chemical inducers either individually or in combination inhibited growth of the tested pathogenic fungi. Biocontrol agents combined with chemical inducers recorded the highest inhibited growth especially in case SA + T. viride and SA + B. megaterium. Under green house and field conditions, all treatments significantly reduced damping-off and root rot/wilt severity and increased of survival plants. Also, these treatments increased fresh and weights of the survival plants in pots compared with control. The combination between biocontrol agents and chemical inducers were more effective than used of them individually and SA + T. viride was the best treatment in this respect. Also, under field conditions, all these treatments significantly increased growth parameters (plant height and number of branches per plant) and yield components (number of pods per plant and number of seeds per plant, weight of 100 seeds and total yield per feddan) and protein content in both seasons (2010~2011 and 2011~2012). Faba bean seeds soaked in SA + T. viride and SA + B. megaterium were recorded the highest growth parameters and yield components. Generally, the combination between biocontrol agents and chemical inducers recorded the best results for controlling damping-off and root rot/wilt diseases in greenhouse and field with addition improved plant growth and increased yield components in field.

Characterization of Two Self-Sufficient Monooxygenases, CYP102A15 and CYP102A170, as Long-Chain Fatty Acid Hydroxylases

  • Rimal, Hemraj;Lee, Woo-Haeng;Kim, Ki-Hwa;Park, Hyun;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.777-784
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    • 2020
  • Self-sufficient P450s, due to their fused nature, are the most effective tools for electron transfer to activate C-H bonds. They catalyze the oxygenation of fatty acids at different omega positions. Here, two new, self-sufficient cytochrome P450s, named 'CYP102A15 and CYP102A170,' from polar Bacillus sp. PAMC 25034 and Paenibacillus sp. PAMC 22724,respectively, were cloned and expressed in E. coli. The genes are homologues of CYP102A1 from Bacillus megaterium. They catalyzed the hydroxylation of both saturated and unsaturated fatty acids ranging in length from C12-C20, with a moderately diverse profile compared to other members of the CYP102A subfamily. CYP102A15 exhibited the highest activity toward linoleic acid with Km 15.3 μM, and CYP102A170 showed higher activity toward myristic acid with Km 17.4 μM. CYP10A170 also hydroxylated the Eicosapentaenoic acid at ω-1 position only. Various kinetic parameters of both monooxygenases were also determined.

Antimicrobial Activity and Characterization of Volatile Flavor EXtracts from Agastache rugosa

  • Song, Jong-Ho;Kim, Min-Ju;Kwon, Kyuk-Dong;Lee, Won-Koo;Park, In-Ho
    • Preventive Nutrition and Food Science
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    • v.4 no.2
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    • pp.97-102
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    • 1999
  • Antimicrobial activity and chemical composition of volatile flavor extracts from Agastache rugosa were investigated. The volatile flavor extracts were obtained from leaves and stems of Agastache rugosa by simultaneous distillation extraction (SDE) method. Antimicrobial activity was investigated by disc diffusion and broth dilution methods against several microorganisms of Bacillus cereus, bacillus megaterium, Bacillus subtilis, Corynebacterium xerosis, Staphylo coccus aureus, Staphylococcus epidermidis, Agrobacterium rhizogenes , Agrobacterium tumefaciences, Enterobacter cloacae, Escherichia coli, Salmonella typhi, Vibrio parahaemolyticus, Candida utilis and Saccharomyces cerevisiae. Volatile flavor extractsfrom leaves have strong antimicrobial activity against C.utilis and S.cerevisiae. When 0.12% volatile flavor extracts from fresh leaves were included in the medium, lag phase of C. utilis was extended 6 hr and that of S.utilis and S.cerevisiae was extended 2hr. Further analyses were performed to elucidatethe effective component of the extracts. The major component of volatile flavor was estragole, a phenolic compound. Minor components were determined to be terpenes , alcohols, acids , esters, ketones and aldethydes.

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Chemical Composition and Antimicrobial Activity of Cone Volatile oil of Cupressus macrocarpa Hartwig from Nilgiris, India

  • Manimaran, S.;Themozhil, S.;Nanjan, M.J.;Suresh, B.
    • Natural Product Sciences
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    • v.13 no.4
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    • pp.279-282
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    • 2007
  • The aim of the present study was to investigate the various chemical components present in the cone volatile oil of Cupressus macrocarpa and also determine its antimicrobial activity. Totally 13 compounds were identified with 99.99% by GC-MS analysis. The major compounds identified were terpinene-4-ol (19.42%), dinopol (15.63%), ${\alpha}$-pinene (13.58%), and ${\beta}$-pinene (12.16%). The antimicrobial activity was carried out for the oil and a 2% cream formulation using cup plate method by measuring the zone of inhibition. The gram positive organisms used were Bacillus subtilis, Staphylococcus aureus, Bacillus megaterium, and Bacillus cogulans. The gram negative organisms used were Escherichia coli, Kleibseilla pneumonia, Pseudomonas aeruginosa and Salmonella typhi. In vitro antifungal studies were also carried out by using organisms, Candida albicans, Aspergillus flavus, Trichoderma lignorum and Cryptococcus neoformans. The standard drugs used were penicillin ($100{\mu}g/mL$), gentamycin ($100{\mu}g/mL$) and griseofulvin ($100{\mu}g/mL$) for gram positive bacteria, gram negative bacteria and fungi respectively. Both oil and cream formulation showed good activity against fungi than bacteria. This study is being reported for the first time on cone volatile oil of this plant.

Characteristics of Soy Protein Hydrolysates with Enzymes Produced by Microorganisms Isolated from Traditional Meju (전통 메주 유래 미생물이 생산하는 효소에 의한 대두단백 분해물의 특성)

  • 정낙현;신용서;김성호;임무현
    • Food Science and Preservation
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    • v.10 no.1
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    • pp.80-88
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    • 2003
  • In order to establish the enzymatic hydrolysis system improving of taste and flavor in the preparation of soy protein hydrolysates using the enzymes with excellent hydrolytic ability and different hydrolysis pattern of soy protein, Degree of hydrolysis(DH) and surface hydrophobicity under the optimal conditions of enzyme reaction, hydrolysis patterns by the SDS electrophoresis and sensory evaluation of soy protein hydrolysates by enzyme reactions were investigated. Four enzyme reactions were highly activated at pH 7.0, 45$^{\circ}C$ under the optimal conditions. As result of changes on the pattern of soy-protein hydrolysates by SDS-electrophoresis, high molecular peptides of hydrolysates by No. 5(Mucor circinelloides M5) and No. 16(Bacillus megaterium B16) enzymes were slowly decrease and 66KD band of these were remained after 3hours reaction. Production of low molecular peptides of hydrolysates by No. 4(Aspergillus oryzae M4) and No. 95(Bacillus subtilis YG 95) enzymes were remarkably detected during the proceeding reactions. As results of HPLC analysis, low molecular peptides of 15∼70KD were mainly appeared during the proceeding enzyme reactions. And, the more DH was increased, the more SDS-surface hydrophobicity was decreased. Hydrolysates by No. 4 enzyme was not only the highest DH of all hydrolysates, but the strongest bitter taste in a sensory evaluation. Sweat taste among the hydrolysates showed little difference. But, when combinative enzymes were treated, combinative enzyme of No. 4(Aspergillus oryzae M4)and No. 16(Bacillus megaterium B16) showed the strongest sweat taste. In conclusion, we assumed that it will be possible to prepare the hydrolysates having functionality when soy-protein were hydrolyzed by these specific enzymes.

Effects of Plants, Rhizobacteria and Physicochemical Factors on the Phytoremediation of Contaminated Soil (오염 토양의 식물상 복원효율에 미치는 식물, 근권세균 및 물리.화학적 인자의 영향)

  • Hong, Sun-Hwa;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.261-271
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    • 2007
  • Phytoremediation is an economic and environmentally friendly technique to remediate contaminated-soil. In this study, the effects of plants, rhizobacteria and physicochemical factors on phytoremediation have been reviewed. For successful phytoremediation, the selection of plants is primarily important. To remediate soil contaminated with petroleum hydrocarbon, raygrass (Lolium multiflorum lam), white mustard, vetch (Vicia villosa), tall fescue (Festuca arundinacea), legumes, poplar, and Pine (Pinus densiflora) were mainly applied, and the removal efficiency of petroleum hydrocarbon were ranged 68 to 99%. Corn (Zea mays), raygrass (Lolium multiflorum lam), vetch (Vicia villosa), mustard, clover (Trifolium repens), and tall fescue (Festuca arundinacea) were used for the removal of polycyclic aromatic hydrocarbon, and their removal efficiencies were 50-98%. Rhizobacteria play significant roles for phytoremediation because they can directly participate in the degradation of contaminant as well as promoting plants growth. The following rhizobacteria were preferred for phytoremediation: Azospirillum lipoferum, Enterobactor cloacae, Azospirillum brasilense, Pseudomonas putida, Burkholderia xenovorans, Comamonas testosterone, Pseudomonas gladioli, Azotobacter chroococcum, Bacillus megaterium, and Bacillus subtilis. Pysicochemical factors such as pH, temperature, nutrient, electron acceptor, water content, organic content, type of contaminants are consequential limiting factors for phytoremediation.

Isolation and Identification of Bacillus sp. with High Protease and Amylase Activity from Sunchang Traditional Kochujang

  • Jung, Sung-Tae;Kim, Min-Hwa;Shin, Dong-Hwa;Kim, Yong-Suk
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.519-526
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    • 2008
  • To improve the quality of traditional kochujang, strains with high protease and amylase activity were isolated and identified from Sunchang traditional kochujang. Twenty-three strains strongly producing protease and 16 strains strongly producing $\alpha$- and $\beta$-amylase were isolated by using 1% isolated soy protein agar medium and 2% starch agar medium, respectively. Protease activities of the IA7, I5, and IA2 strain were 22.5, 21.2, and 20.6 unit/mL, respectively, and were higher than those of the other strains. Stains with high $\alpha$-amylase activity included K9 (967.8 unit/mL), K14 (828.3 unit/mL), K13 (662.5 unit/mL), K8 (601.5 unit/mL), and K11 (405.9 unit/mL). The $\beta$-amylase activity of the K11 strain was the highest, 34.3 unit/mL, among the isolated strains. Based on morphological, physiological properties, and API 50CHB-kit test for assimilation of 49 carbohydrates, 8 strains selected according to protease, $\alpha$-amylase, and $\beta$-amylase activities were tentatively identified as Bacillus megaterium (IA2), Bacillus subtilis (IA7, 15), Bacillus amyloliquefaciens (K8, K9, K11, and K13), and Bacillus stearothermophillus (K14). The IA7, 15, and K11 strains were finally identified as B. subtilis (99% ID) based on 16S rDNA sequencing.

Crystal Structure and Functional Characterization of a Cytochrome P450 (BaCYP106A2) from Bacillus sp. PAMC 23377

  • Kim, Ki-Hwa;Lee, Chang Woo;Dangi, Bikash;Park, Sun-Ha;Park, Hyun;Oh, Tae-Jin;Lee, Jun Hyuck
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1472-1482
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    • 2017
  • Bacterial cytochrome P450 (CYP) steroid hydroxylases are effectively useful in the pharmaceutical industry for introducing hydroxyl groups to a wide range of steroids. We found a putative CYP steroid hydroxylase (BaCYP106A2) from the bacterium Bacillus sp. PAMC 23377 isolated from Kara Sea of the Arctic Ocean, showing 94% sequence similarity with BmCYP106A2 (Bacillus megaterium ATCC 13368). In this study, soluble BaCYP106A2 was overexpressed to evaluate its substrate-binding activity. The substrate affinity ($K_d$ value) to 4-androstenedione was $387{\pm}37{\mu}M$. Moreover, the crystal structure of BaCYP106A2 was determined at $2.7{\AA}$ resolution. Structural analysis suggested that the ${\alpha}8-{\alpha}9$ loop region of BaCYP106A2 is intrinsically mobile and might be important for initial ligand binding. The hydroxyl activity of BaCYP106A2 was identified using in vitro enzyme assays. Its activity was confirmed with two kinds of steroid substrates, 4-androstenedione and nandrolone, using chromatography and mass spectrometry methods. The main products were mono-hydroxylated compounds with high conversion yields. This is the second study on the structure of CYP106A steroid hydroxylases, and should contribute new insight into the interactions of bacterial CYP106A with steroid substrates, providing baseline data for studying the CYP106A steroid hydroxylase from the structural and enzymatic perspectives.