• Title/Summary/Keyword: Bacillus subtilis

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Bioactivity of Metabolites from Actinomycetes Isolates from Red Sea, Egypt

  • Osman, Mohamed E.;El-nasr, Amany A. Abo;Hussein, Hagar M;Hamed, Moaz M
    • Microbiology and Biotechnology Letters
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    • v.50 no.2
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    • pp.255-269
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    • 2022
  • Actinomycetes isolated from marine habitats represent a promising source of bioactive substances. Here, we report on the isolation, identification, productivity enhancement and application of the bioactive compounds of Streptomyces qinglanensis H4. Eighteen marine actinomycetes were isolated and tested for resistance to seven bacterial diseases. Using 16S rRNA sequencing analysis (GenBank accession number MW563772), the most powerful isolate was identified as S. qinglanensis. Although the strain produced active compound(s) against a number of Gram-negative and Gram-positive bacteria, it failed to inhibit pathogenic fungi. The obtained inhibition zones were 22.0 ± 1.5, 20.0 ± 1, 16.0 ± 1, 12.0 ± 1, 22.0 ± 1 and 24.0 ± 1 mm against Bacillus subtilis ATCC 6633, Escherichia coli ATCC 19404, Enterococcus faecalis ATCC 29212, Pseudomonas aeruginosa ATCC 9027, Candida albicans ATCC 10231 and Staphylococcus aureus ATCC6538, respectively. To maximize bioactive compound synthesis, the Plackett-Burman design was used. The productivity increased up to 0.93-fold, when S. qinglanensis was grown in optimized medium composed of: (g/l) starch 30; KNO3 0.5; K2HPO4 0.25; MgSO4 0.25; FeSO4·7H2O, 0.01; sea water concentration (%) 100; pH 8.0, and an incubation period of 9 days. Moreover, the anticancer activity of S. qinglanensis was tested against two different cell lines: HepG2 and CACO. The inhibition activities were 42.96 and 57.14%, respectively. Our findings suggest that the marine S. qinglanensis strain, which grows well on tailored medium, might be a source of bioactive substances for healthcare companies.

Screening of Bacterial Antagonists to Develop an Effective Cocktail against Erwinia amylovora

  • Choi, Dong Hyuk;Choi, Hyun Ju;Kim, Yeon Ju;Lim, Yeon-Jeong;Lee, Ingyeong;Park, Duck Hwan
    • Research in Plant Disease
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    • v.28 no.3
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    • pp.152-161
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    • 2022
  • Several types of chemical bactericides have been used to control fire blight. However, their excessive usage leads to environmental deterioration. Therefore, several researchers have analyzed antagonistic microorganisms as promising, effective, and safe biological control agents (BCAs). The primary aim of this study was to screen for potential antagonistic bacteria that suppress Erwinia amylovora. Among the 45 isolates studied, 5 strains showed the largest inhibition zone against E. amylovora. 16S rRNA gene sequencing identified them as Bacillus amyloliquefaciens (KPB 15), B. stratosphericus (KPB 21), B. altitudinis (KPB 25), B. safensis (KPB 31), and B. subtilis (KPB 39). KPB 25 and 31 reduced the lesion size of fire blight by 50% in immature apple fruits, and did not show antagonism against each other. Therefore, KPB 25 and 31 were selected to develop an antagonistic mixture against fire blight. Although the mixture with KPB 25 and 31 showed a slightly increased ability to reduce lesion size on immature fruits, they did not exhibit a synergistic effect in reducing E. amylovora population compared to each strain alone. Nevertheless, we have identified these two strains as useful and novel BCAs against fire blight with additional benefits safety and potential in developing a mixture without loss of their activity, owing to the absence of antagonism against each other.

Inhibition of Pathogenic Bacteria and Fungi by Natural Phenoxazinone from Octopus Ommochrome Pigments

  • Lewis-Lujan, Lidianys Maria;Rosas-Burgos, Ema Carina;Ezquerra-Brauer, Josafat Marina;Burboa-Zazueta, Maria Guadalupe;Assanga, Simon Bernard Iloki;del Castillo-Castro, Teresa;Penton, Giselle;Plascencia-Jatomea, Maribel
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.989-1002
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    • 2022
  • Cephalopods, in particular octopus (Octopus vulgaris), have the ability to alter their appearance or body pattern by showing a wide range of camouflage by virtue of their chromatophores, which contain nanostructured granules of ommochrome pigments. Recently, the antioxidant and antimicrobial activities of ommochromes have become of great interest; therefore, in this study, the pH-dependent redox effect of the extraction solvent on the antioxidant potential and the structural characterization of the pigments were evaluated. Cell viability was determined by the microdilution method in broth by turbidity, MTT, resazurin, as well as fluorescence microscopy kit assays. A Live/Dead Double Staining Kit and an ROS Kit were used to elucidate the possible inhibitory mechanisms of ommochromes against bacterial and fungal strains. The results obtained revealed that the redox state alters the color changes of the ommochromes and is dependent on the pH in the extraction solvent. Natural phenoxazinone (ommochromes) is moderately toxic to the pathogens Staphylococcus aureus, Bacillus subtilis, Salmonella Typhimurium and Candida albicans, while the species Pseudomonas aeruginosa and Pseudomonas fluorescens, and the filamentous fungi Aspergillus parasiticus, Alternaria spp. and Fusarium verticillioides, were tolerant to these pigments. UV/visible spectral scanning and Fourier- transform infrared spectroscopy (FTIR) suggest the presence of reduced ommatin in methanol/ HCl extract with high intrinsic fluorescence.

Studies on Biological Activities of Woody Essential Oils( I ) - Antimicrobial Activity of Essential Oils from Coniferous Needles - (수목 정유의 생리활성에 관한 연구( I ) - 침엽수 잎 정유의 항균활성 -)

  • Lee, Sung-Suk;Kang, Ha-Young;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.30 no.1
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    • pp.48-55
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    • 2002
  • The essential oils were extracted from the needles of several softwood tree species, Pinus densiflora, Pinus koraiensis, Chamaecyparis obtusa and Chamaecyparis pisifera to investigate their biological activities. The antifungal activity by hyphal growth inhibition method was determined by three wood rot fungi, one mouldous fungi, and three tree pathogenic fungi and the antibacterial activity by paper disc method and turbidiometric method by three gram positive bacteria and one gram negative bacteria, Escherichia coli, were also tested. Antifungal activity was the best in the essential oil from C. obtusa, and the following was resulted in order of P. densiflora and P. koraiensis. The essential oil from P. densiflora showed higher antifungal activity against wood rot fungi than that from P. koraiensis while the result was contrary against tree pathogenic fungi. However, the antibacterial activity was the best in the essential oil from P. densiflora, and the following was in order of C. obtusa and P. koraiensis. From these results, the variation of biological activity seemed to be due to the chemical composition of essential oils.

Changes in Physiological Activity of Gardenia Fructus by Roasting Treatment

  • Park, Ji Sun;Choi, Ha Kyoung;Kang, Jeong Eun;Shin, Yong Wook;Lee, In Ah
    • Journal of People, Plants, and Environment
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    • v.23 no.4
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    • pp.455-464
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    • 2020
  • Background and objective: This study was conducted to examine changes in the composition and physiological activity of Gardenia Fructus after being roasted. Methods: The antioxidant, anti-inflammatory and antibacterial activity of Gardenia Fructus was evaluated using the Gardenia Fructus (GF) and roasted Gardenia Fructus (RGF) ethanol extracts, and their components were analyzed through HPLC. Results: As a result, it was confirmed that the content of gardenoside and geniposide decreased and the content of genipin increased when GF was roasted. The total content of polyphenols was 54.5 ± 2.18 mg gallic acid equivalents (GAE) per gram of the GF extract and 69.6 ± 0.36 mg GAE per gram of the RGF extract. As a result of evaluating 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, both the GF and RGF extracts showed the similar activity to ascorbic acid at the concentrations of 1 mg/mL or higher. In RAW 264.7 macrophages stimulated by lipopolysaccharides (LPS), the RGF extract showed a higher effect of reducing NO production, and significantly reduced the expression of an inflammatory cytokine, IL-6. As a result of evaluating the antimicrobial activity, the RGF extract showed higher antimicrobial activity against Escherichia coli and Bacillus subtilis. In the dextran sulfate sodium salt (DSS) induced inflammatory bowel disease mouse model, the RGF extract reduced the weight of the spleen, and both the GF and RGF extracts reduced the number of bacteria in the colon. Conclusion: Therefore, it has been confirmed through this study that roasting at a high temperature changes the main components of the GF extract and increases its biological activity. The RGF extract is expected to be used as a natural material with antioxidant, anti-inflammatory and antibacterial effects.

Biological Control of Sesame Soil-born Disease by Antifungal Microorganisms (참깨 토양전염성병(土壤傳染性病)의 생물학적방제(生物學的防除))

  • Shin, G.C.;Im, G.J.;Yu, S.H.;Park, J.S.
    • Korean journal of applied entomology
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    • v.26 no.4 s.73
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    • pp.229-237
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    • 1987
  • In order to study the biological control of soil-borne disease of sesame, antagonistic isolates of Trichoderma , Bacillus sand streptomyces to Fusarium oxysporum and Rhizoctonia solani were isolated from the rhizosphere soils of sesame plants and some other habitats. Out of the isolates of microorganisms collected a strain of Trichoderma viride was selected as a biological control agent for the study and its effect on the control of damping-off and the seedling growth of sesame was investigated. The results obtained are as follows: 26 percents of Bacillus spp. isolated from the rhizosphere soil of sesame plants showed antagonism to two pathogenic fungi. Important species were B. Subtilis and B. polymyxa. Streptomyces species isolated from the rhizosphere soils of sesame lysed the cell wall of hyphae and conidia of F. oxysporum and reduced conspicuously the formation of macroconidia and chlamydospores of the fungus. 84 percents of Trichoderma spp. isolated from the rhizosphere soil of sesame plants were antagonistic to F. oxysporum and 60 percents of the isolates were antagonistic to both F. oxysporum and R. solani. Trichoderma viride TV-192 selected from antagonistic isolates of Trichoderma spp. was highly antagonistic to F. oxysporum and soil treatment with the isolate reduced notably damping-off of sesame. T. viride TV-192 showed better growth in crushed rice straw, barley straw and sawdust media than F. oxysporum. Sawdust was selective for the growth of T. viride. Supplementation of wheat bran and mixtures of wheat bran and sawdust inoculated with T. viride TV-192 in the soil reduced remarkably damping-off of sesame by F. oxysporum but high density of the fungus TV-192 caused the inhibition of seed germination and seedling growth of sesame. Inhibitory effects of Trichoderma species on seed germination and seedling growth of sesame were different according to the isolates of the fungus. Normal sesame seedlings on the bed treated with the fungus showed better growth than not treated seedlings.

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Microbiological Studies of Korean Native Soy-sauce Fermentation: A Study on the MicroBora of Fermented Korean Maeju Loaves (한국 재래식간장의 발효미생물에 관한 연구 -한국재래식메주에 발효미생물군에 대하여-)

  • Cho, Duck-Hiyon;Lee, Woo-Jin
    • Applied Biological Chemistry
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    • v.13 no.1
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    • pp.35-42
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    • 1970
  • Five samples of Korean native Maeju(fermented soy-bean mash) loaves which were collected each from Kyunggi, Chungchung, Kangwon, Cholla and Kyungsang-Do were examined for their fermenting microorganisms. The results of taxonomic and ecological studies of fermentation microorganisms in these Maeju loaves were as the fellows. (1) The fungus flora grew only is the outer layer of Maeju loaves. Miscellaneous molds, 3 species of Mucor, 2 species of Pericallium., one species each of Scopulariopsis and Aspergillus, were isolated. None of them seemed exclusively predominant to be able to designate as the ecologically significant. (2) The bacterial flora which consisted of two species, Bacillus subtilis and Bacillus pumilus were distributed uniformly in th a entire Maeju loaves. The inner parts of Maeju loaves were especially inhabited solely by these bacterial flora. Probably the Korean native Maeju fermentation could be characterized by these bacterial flora. A Staphylococcus species was also isolated probably as a casual contaminant. (3) The yeasts, Rhodotorula flava and Torulopsis dattila, were isolated from Maeju loaves though their ecological significance was not clear. (4) The ecological aspects of fermentation microbes in the outer and inner parts of Maeju loaves were apparently different, consequently different fermentation processes might have occurred in these two parts and it brought quite different final outlooks in the final matured Maeju loaves. The outer part, rather rigid and dry, retained the light brown color of boiled soy-bean; whereas the inner part, soft and sticky, showed dark brown color indicating severe chemical changes. (5) The aflatoxin producing mold, Aspergillus oryzae was isolated from one sample among 5 of Maeju loaves. In addition to the low probability of isolability from Maeju loaves samples, since this mold grew only in the outer layer of Maeju loaves with such a low population density, about $10^4/g$, perhaps the aflatoxin problem in Korean native soysauce may not be critical.

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Effects of Different Microbial Culture Supplements on In vitro and In situ Ruminal Fermentation Characteristics of Italian ryegrass Silage (미생물제 처리에 의한 이탈리안 라이그라스 사일리지의 In vitro 및 In situ 반추위 발효특성에 미치는 영향)

  • Lim, D.H.;Ki, K.S.;Choi, S.H.;Kim, T.I.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.4
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    • pp.309-317
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    • 2016
  • The study was conducted to evaluate the effects of microbial culture supplements on ruminal fermentation and fermentative quality of Italian ryegrass silage (IRGS) both in vitro and in situ. Three species of microbes (Lactobacillus casei (LC), Bacillus subtilis (BS), and Saccharomyces cerevisiae (SC)) were used in this study. They were applied to IRGS at 30 days after silage manufacture. Various items were measured using in vitro and in situ incubation technique after each microbial supplement was inoculated into IRGS at $0.5{\times}10^4CFU/g$. In the first experiment, in vitro ruminal fermentation characteristics of IRGS were evaluated at 0, 12, 24, 48, and 72 hours after microbes were inoculated into IRGS. In the second experiment, in situ fermentation characteristics were investigated at 0, 1, 3, and 5 days after the inoculation of each microbial supplement. In vitro ruminal $NH_3-N$ content was significantly (p<0.05) increased in LC-, BS-, and SC-IRGS at 12 hrs post incubation compared to that in control IRGS. In vitro ruminal total VFA concentration and dry matter digestibility (DMD) of IRGS were not significantly difference among LC-, BS-, and SC-IRGS, although they were numerically increased in LC-IRGS than those of the other IRGS. In addition, this study evaluated the fermentation characteristics and in situ DMD of IRGS with the lapse of incubation time up to 5 days. Throughout the incubation times from 1 day to 5 days, the pH value was significantly (p<0.05) lower in BS-, LC-, and SC-IRGS than that in control IRGS. Lactate was significantly (p<0.05) higher, and significantly (p<0.05) butyrate was lower in LC-IRGS than that in other treatments at 0 day. It was higher (p<0.05) in control IRGS than that of BS-, LC-, and SC-IRGS at 1-5 days. In situ DMD tended to increase in BS-, LC-, and SC-IRGS compared to that in control IRGS. Especially, DMD was higher in SC-IRGS than that in other treatments at 0 day. It tended to be higher in LC-IRGS at all incubation time. Taken together, these results suggest that it might be useful to select a microorganism by considering the feeding time of IRGS to ruminants because organic acids and DMD of IRGS were affected by the incubation time of each microorganism with IRG silage, especially for L. casei decreased the content of acetate and butyrate in IRGS.

Anti-Obesity Effects of Fermented Soybean Oils in 3T3-L1 Pre-Adipocytes and High Fat Diet-Fed C57BL/6J Mice (발효콩 유지의 3T3-L1 지방전구세포와 고지방식이를 급여한 C57BL/6J 생쥐에 대한 항비만 효과)

  • Kim, Seon-Woong;Kim, Nam-Seok;Oh, Mi-Jin;Kim, Ha-Rim;Kim, Min-Sun;Lee, Da-Young;Yoon, Suk-Hoo;Jung, Mun-Yhung;Kim, Hun-Jung;Lee, Chang-Hyun;Oh, Chan-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.3
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    • pp.279-288
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    • 2017
  • This study investigated the manufacturing of fermented soybean oil using a fermenting strain commonly processed for soybeans [Bacillus amyloliquefaciens (BA), Bacillus subtilis (BS), Lactobacillus acidophilus (LBA), and B. subtilis+L. acidophilus (BLO)] and evaluated its anti-obesity activities. Cytotoxicity of four kinds of fermented soybean oils was not observed in 3T3-L1 preadipocytes at 10 and $50{\mu}g/mL$. Triglyceride content was reduced by 20.6% in the BLO group at a treatment concentration of $50{\mu}g/mL$. The simultaneous treatment of fermented soybean oil and differentiation induction medium decreased $PPAR{\gamma}$ and $C/EBP{\alpha}$ gene expression at a concentration of $50{\mu}g/mL$ and blocked adipocyte differentiation by increasing adiponectin gene expression. The inhibitory effect of adipocyte differentiation was greatest in the BLO group. C57BL/6J mice were examined for 4 weeks after being separated into seven groups [normal diet group (N), high fat diet group (C), group fed high fat diet combined with regular soybean oil (SO), group fed non-fermented soybean oil (NF), and groups fed high fat diet combined with 5% fermented soybean oil (BA, BS, LBA, and BLO)] to identify the effects of soybean oil on body weight, serum lipid, adiponectin, insulin, and leptin levels in mice with high fat diet-induced obesity. The body weight and serum lipid level of the C group increased drastically compared to those of the N group. In contrast, the group fed a diet combined with fermented soybean oil showed decreases in weight, serum total cholesterol, LDL-cholesterol, and triglyceride levels compared to those of the C group. Moreover, soybean oil was found to be effective in the BLO group. In conclusion, fermented soybean oil has positive effects in prohibiting adipocyte differentiation increased by high fat diet and improving serum lipid composition. Therefore, fermented soybean oil can be used as a functional food material with anti-obesity activity.

Development of Odor-Reduced Korean Traditional Cheonggukjang Added with Job's Tears (냄새저감형 율무청국장 제조에 관한 연구)

  • Park, Ju-Hun;Han, Chan-Kyu;Choi, Sook-Hyun;Lee, Bog-Hieu;Lee, Hye-Jeong;Kim, Sung-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.2
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    • pp.259-266
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    • 2011
  • This study was performed to develop an odor-reduced Korean traditional Cheonggukjang (fermented soybean paste) by adding Job's tears to Cheonggukjang made with soybean (control) only. The study included the determination of best addition percentage of Job's tears versus soybean, sensory evaluation, and aroma patterns and its components as well as proximate composition analyses of Cheonggukjang. Job's tears Cheonggukjang (CAJT) was prepared by inoculating Bacillus subtilis and fermented at $40^{\circ}C$ for 48 hours. Ratios of soybean : Job's tears for Job's tears Cheonggukjang were 1:1, 2:1, 3:1, and 4:1, respectively. In comparison of proximal composition, Job's tears Cheonggukjang was high in moisture and carbohydrates, but low in calorie, fat, protein, ash and amino nitrogen. The pH of CAJT was lower than those of control and conventional Cheonggukjang (CC) sold in the market, and the acidity was the lowest in CC. The color of Job's tears Cheonggukjang appeared the whiter and more yellowish, but less reddish in proportion to contents of Job's tears. The contents of viscous substances were higher in CAJT compared with those of control and CC. The contents of glutamic acid, the major components of viscous substances were lower in CAJT than in control and CC, and the contents of fructose higher in CAJT. The scores of sensory evaluation were the highest in Job's tears Cheonggukjang mixed with soybean : Job's tears=4:1. The aroma pattern of CC analyzed was conspicuously dissimilar to the control and CAJT and also discriminated by electronic nose examination. The pyrazines, volatile compounds peculiar to Cheonggukjang, were found to be lower in CAJT by SPME-GC/MS assay. Additionally, the acetic acid, butanoic acid, and naphthalene causing off-flavor were identified in CC, but not in Job's tears Cheonggukjang. The strength of odor through sensory evaluation was by far the lowest in CAJT among the groups. From the findings, it had shown that Job's tears Cheonggukjang would be produced successfully when the ratio of 4:1 (soybean : Job's tears) is employed. Also, it was proved that appropriate ratio of Job's tears addition makes Cheonggukjang odor-reduced and well accepted by people.