• Title/Summary/Keyword: broth culture

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Characterization of Bacillus licheniformis KJ-9 Isolated from Soil (토양으로부터 분리한 Bacillus licheniformis KJ 9의 특성)

  • Seo, Dong-Cheol;Ko, Jeong-Ae;Gal, Sang-Won;Lee, Sang-Won
    • Journal of Life Science
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    • v.20 no.3
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    • pp.403-410
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    • 2010
  • In order to produce high-quality fermenting composts, a microorganism was isolated from the natural world. The bacterium has not only in high enzyme activities but also had good antimicrobial activities against phytopathogenic microorganisms. Its cultivating characteristics were then investigated. Bacterium KJ-9, which contains high CMCase, protease and chitinase activities and excellent antimicrobial activities against phytopathogenic microorganisms, was separated from leaf mold and identified as Bacillus licheniformis by two methods: Bergey's Manual of Systematic Bacteriology and API 50 CHL Carbohydrate Test Kit (Bio Merieux, France) using an ATB (Automated Identification) computer system (Bio Merieux, France). Optimal medium for cultivation of B. licheniformis was 2% soluble starch as a carbon source, 0.5% yeast extract as a nitrogen source and 0.05% $MgSO_4{\cdot}7H_2O$. Optimal growth conditions of pH, temperature and shake speed were pH 7.0, $50^{\circ}C$ and 180 rpm, respectively. Culture broth of B. licheniformis KJ-9 cultured for 36~60 hr was effective in fungicidal activities against plant pathogens including Botrytis cinerea, Corynespora cassicola, Fusarium oxysporum, and Rhizoctonia solani.

Plant Growth Promotion by Purple Nonsulfur Rhodopseudomonas faecalis Strains (자색비유황세균 Rhodopseudomonas faecalis의 식물생장촉진능)

  • Lee, Eun-Seon;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.157-161
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    • 2010
  • Photosynthetic purple nonsulfur bacterial strains were isolated from the sediments collected from rice paddy fields and sludges of wastewater treatment plant, and their plant growth promoting capabilities were examined. Most well known phytohormones, auxin such as indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) and 5'-aminolevulinic acid (ALA) were detected by HPLC in the culture broth of these isolates. Among the isolated bacteria, Rhodopseudomonas faecalis D15 showed the highest production rate of 769.8 ${\mu}g$/mg protein of IAA, 1323 ${\mu}g$/mg protein of IBA and 7.4 mM/mg protein of ALA in the modified Biebl and Pfennig's medium. R. faecalis C9 showed the highest production rate of 20.82 ${\mu}g$/mg protein of gibberellin. In consequence, the root length and dry weight of the germinated tomato seedling treated with R. faecalis isolates were longer and heavier than those of uninoculated control after 15 days of incubation in the soil. Especially, the dry weight of germinated tomato seedling increased by 119.4% in C9-treated samples after 15 days. These purple nonsulfur bacteria may be utilized as environment-friendly biofertilizer in the agriculture.

Effect of Tyrosinase Inhibitors on the Melanogenesis of Gold Fish(Jet Black Color) (Tyrosinase 저해제가 검은툭눈붕어의 멜라닌 생성에 미치는 영향)

  • Han, Dae-Seok;Jung, Sung-Won;Kim, Seok-Joong;Kim, Sang-Hee;Ahn, Byung-Hak
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.1089-1094
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    • 1996
  • The in vivo effect of tyrosinase inhibitors in the melanogenesis of gold fish (jet black color) was evaluated by measuring surface color and observing melanin pigment. The fish was firstly cultivated in 0.9% NaCl solution for 1 week to induce melanogenesis, and then, it was transferred to each treatment group containing tyrosinase inhibitor. The fish was grouped into control. food additive group (addition of 5 mM glutathione, 5 mM cysteine, and 1 mM benzoic acid), microbial inhibitor group (addition of culture broth of Aspergillus oryzae in shiitake and glucose medium), and plant extract group (addition of the mixed extracts of green tea, beet, red chicory, and nameko). After 6 days, the fish was anesthetized by electric shock, and color of pectoral region, lateral region, and dorsal fin was measured. Hunter's L and b values of treated group were generally higher than those of control group, indicating that the tyrosinase inhibitors could inhibit the melanogenesis of the fish. Effect of plant extract was apparent, though relatively weak, not because it did not work in vivo, but because a sufficient amount of extract could not be added to fish globes. If a large amount of extract was added, fish gradually died due to a microbial contamination. Microscopic observation of melanin in lateral scale and dorsal fin showed that in the treated groups with tyrosinase inhibitors, the number of melanophore per unit area and the size of one melanophore decreased.

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Effect of Cellular Phospholipase A2 Inhibition on Enhancement of Bt Insecticidal Activity (세포성 인지질분해효소 활성 억제에 따른 비티 살충력 증가 효과)

  • Eom, Seonghyeon;Park, Jiyeong;Kim, Kunwoo;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.53 no.3
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    • pp.271-280
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    • 2014
  • Some bacterial metabolites of Xenorhabdus nematophila (Xn) inhibit phospholipase $A_2$ ($PLA_2$) activity to shutdown eicosanoid biosynthesis in target insects. However, little has been known about the target insect $PLA_2$ of these bacterial metabolites. Eight bacterial metabolites identified in Xn culture broth exhibited significant insecticidal activities against larvae of both lepidopteran species of Plutella xylostella and Spodoptera exigua. Moreover, these bacterial metabolites significantly enhanced insecticidal activities of Bacillus thuringiensis (Bt). To determine target $PLA_2$, we cloned and over-expressed cellular $PLA_2$ ($SecPLA_2$) of S. exigua. Purified $SecPLA_2$ catalyzed phospholipids derived from the fat body and released several polyunsaturated fatty acids. Most Xn metabolites significantly inhibited $SecPLA_2$ activity, but were different in their inhibitory activities. There was a positive correlation between the inhibition of $SecPLA_2$ and the enhancement of Bt insecticidal activity. These results indicate that $SecPLA_2$ is a molecular target inhibited by Xn metabolite.

Immobilization of Bacillus sp. Strains, Catalase Producing Bacteria and Their Hydrogen Peroxide Removal Characteristics (카탈라제를 생산하는 고초균 (Bacillus sp.)의 고정화 및 과산화수소 분해 특성)

  • Han, Kyung-Ah;Jang, Yun-Hee;Rhee, Jong-Il
    • KSBB Journal
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    • v.25 no.6
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    • pp.520-526
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    • 2010
  • In this work we have investigated the production of catalase from Bacillus sp. strains, which were screened and identified from soil. These strains were cultivated in shaking flasks with tryptic soy broth (TSB) at $30^{\circ}C$ and 200 rpm. Effects of the temperature and pH on the stability of the native catalase and whole cell viability were studied in the temperature range of $25-60^{\circ}C$ and the pH range of 7-13. Korean natural zeolite was added to culture medium and mixed with microorganisms for 24 hours. The native catalase maintained its activity over $50^{\circ}C$. The enzyme acitiviy of the catalase from Bacillus flexus BKBChE-3 was highest among the Bacillus sp. strains studied. Bacillus flexus BKBChE-3 and immobilized Bacillus cells have survived under extreme conditions of over $50^{\circ}C$ and pH 12. 60 mL of 10.5 mM $H_2O_2$ solution were entirely removed within 1 hour with catalase produced from Bacillus sp. on the flask. When Bacillus cells were immobilized on Korean natural zeolite, colony forming unit of Bacillus flexus BKBChE-3 was increased and high efficiency of hydrogen peroxide removal was observed.

Isolation and Characterization of Plant Growth Promoting Rhizobacterium Bacillus subtilis YK-5 from Soil (토양에서 식물생육촉진 활성을 가진 균주 Bacillus subtilis YK-5의 분리 및 특성)

  • Yeo, Soo-Hwan;Yook, Young-Min;Kim, Hyun-Soo
    • KSBB Journal
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    • v.24 no.4
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    • pp.334-340
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    • 2009
  • For the isolation of a plant growth-promoting rhizobacteria, strain YK-5 was selected from approximately 400 thermostable strains isolated from special soil samples. Strain YK-5 produced an antifungal compound, and optimum carbon and nitrogen sources for the production of the antifungal compound were investigated against Aspergillus flavus as a test strain. Modified LB medium containing 1% peptone, 1% yeast extract and 5% black sugar was determined to be the optimal medium for growth and antifungal compound production. Culture broth of strain YK-5 potently inhibited growth of the phytopathogenic fungus Fusarium oxysporum KACC 40052 for 7 days. The plant growth-promotion function of strain YK-5 was tested against radish and rice in pot trials. Leaf number, plant height and root length in YK-5-treated radish markedly exceeded (> 60%) those of untreated radish. Leaf length and white rootlet development were markedly more prominent than in commercially-treated rice plants. Strain YK-5 was determined to be Bacillus subtilis YK-5 by physiological, chemotaxonomical, and phylogenetical analyses.

Antimicrobial Activity of Garlic Juice against Escherichia coli O157:H7 (마늘즙의 Escherichia coli O157:H7에 대한 항균작용)

  • Kim, Myung-Hee;Kim, So-Young;Shin, Weon-Sun;Lee, Jun-Soo
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.752-755
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    • 2003
  • The antimicrobial activity of fresh garlic juice against Escherichia coli O157:H7 was investigated. When E. coli O157:H7 was cultured for 18 hr in the trypticase soy broth containing 1%, 3%, and 5% garlic juice, viable cell number of E. coli O157:H7 was reduced to $2.3{\times}10^2\;CFU/mL$ at 5% from $7{\times}10^8\;CFU/mL$ at the non-treated culture, respectively. The inhibitory effects of the ground beef treated with 3%, 6%, and 10% garlic juice against E. coli O157:H7 was significantly enhanced with approximate 2 log-reduction compared to that of ground beef without garlic. There was no significant difference in the inhibition of E. coli O157:H7 among the groups with different amounts of garlic juice (p<0.05). These results suggest that garlic juice may function well as a natural preservative in food system.

Establishing Effective Screening Methodology for Novel Herbicide Substances from Metagenome (신규 제초활성 물질 발굴을 위한 메타게놈 스크리닝 방법 연구)

  • Lee, Boyoung;Choi, Ji Eun;Kim, Young Sook;Song, Jae Kwang;Ko, Young Kwan;Choi, Jung Sup
    • Weed & Turfgrass Science
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    • v.4 no.2
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    • pp.118-123
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    • 2015
  • Metagenomics is a powerful tool to isolate novel biocatalyst and biomolecules directly from the environmental DNA libraries. Since the metagenomics approach bypasses cultivation of microorganisms, un-cultured microorganisms that are majority of exists can be the richest reservoir for natural products discovery. To discover novel herbicidal substances from soil metagenome, we established three easy, simple and effective high throughput screening methods such as cucumber cotyledon leaf disc assay, microalgae assay and seed germination assay. Employing the methods, we isolated two active single clones (9-G1 and 9-G12) expressing herbicidal activity which whitened leaf discs, inhibited growth of microalgae and inhibited root growth of germinated Arabidopsis seeds. Spraying butanol fraction of the isolated active clones' culture broth led to growth retardation or desiccation of Digitalia sanguinalis (L) Scop. in vivo. These results represent that the screening methods established in this study are useful to screen herbicidal substances from metagenome libraries. Further identifying molecular structure of the herbicidal active substances and analyzing gene clusters encoding synthesis systems for the active substances are in progress.

Screening of Ochratoxin A Producing Fungi from Greenhouse Horticulture (시설원예산물로부터 Ochratoxin A 생성 곰팡이의 검색)

  • Kang, Sung-Jo;Park, Bong-Jung;Lee, Jong-Ok;Kang, Jin-Soon;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1415-1419
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    • 1998
  • In order to evaluate the safety of greenhouse horticultures, a large number sample sources were collected, and the fungi of Aspergillus sp. and Penicillium sp. were isolated from them. Indirect competitive ELISA method and high performance liquid chromatography (HPLC) were applied to confirm the ochratoxin A producing abilities of isolated strains. One hundred ninety two sample sources including soil, pepper, strawberry and water mellon were collected for fungi isolation from western Gyeongnam, Andong and Gyeongbok. One hundred forty two strains of Aspergillus sp. and one hundred fifty three strains of Penicillium sp. were isolated respectively from them. The isolated fungi were tested for the production of ochratoxin A by ELISA. After culture of them on the modified sucrose low salt medium at $28^{\circ}C$ for 15 days, we found that five strains of Penicillium sp. produced ochratoxin A at the levels of $0.084{\sim}2.128\;{\mu}g/mL$. Among them, #129-2 strain isolated from water melon, showed the highest level of ochratoxin A as $2.128\;{\mu}g/mL$ broth. However, all of isolated Aspergillus sp. didn't produce ochratoxin A. When we compared the results of ELISA method with HPLC method, ochratoxin A production of each isolated strains showed very similar levels.

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Nematicidal Activity of Kojic Acid Produced by Aspergillus oryzae against Meloidogyne incognita

  • Kim, Tae Yoon;Jang, Ja Yeong;Jeon, Sun Jeong;Lee, Hye Won;Bae, Chang-Hwan;Yeo, Joo Hong;Lee, Hyang Burm;Kim, In Seon;Park, Hae Woong;Kim, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1383-1391
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    • 2016
  • The fungal strain EML-DML3PNa1 isolated from leaf of white dogwood (Cornus alba L.) showed strong nematicidal activity with juvenile mortality of 87.6% at a concentration of 20% fermentation broth filtrate at 3 days after treatment. The active fungal strain was identified as Aspergillus oryzae, which belongs to section Flavi, based on the morphological characteristics and sequence analysis of the ITS rDNA, calmodulin (CaM), and β-tubulin (BenA) genes. The strain reduced the pH value to 5.62 after 7 days of incubation. Organic acid analysis revealed the presence of citric acid (515.0 mg/kg), malic acid (506.6 mg/kg), and fumaric acid (21.7 mg/kg). The three organic acids showed moderate nematicidal activities, but the mixture of citric acid, malic acid, and fumaric acid did not exhibit the full nematicidal activity of the culture filtrate of EML- DML3PNa1. Bioassay-guided fractionation coupled with 1H- and 13C-NMR and EI-MS analyses led to identification of kojic acid as the major nematicidal metabolite. Kojic acid exhibited dose-dependent mortality and inhibited the hatchability of M. incognita, showing EC50 values of 195.2 μg/ml and 238.3 μg/ml, respectively, at 72 h post-exposure. These results suggest that A. oryzae EML-DML3PNa1 and kojic acid have potential as a biological control agent against M. incognita.