• Title/Summary/Keyword: Soil microbiology

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Biodegradation of petroleum hydrocarbons by bacteria with surfactant producing capability and cell surface hydrophobicity (계면활성제 생성능과 세포 표면 소수성을 가진 세균 균주들에 의한 석유탄화수소의 생분해)

  • Kwon, Sun-Lul;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.265-272
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    • 2017
  • Some bacteria with different mechanisms for hydrocarbon degradation were isolated from oil-contaminated soils in Korea. Isolate Acinetobacter calcoaceticus SL1 showed biosurfactant- producing activity in oil-spreading test, and it exhibited a good emulsifying activity of 43.6 and 54.5% for diesel oil and n-hexane, respectively. It also has high cell surface hydrophobicity which can make it easily attaches to hydrocarbons and degrade them. It degraded 100% of 1,000 mg/L of n-octadecane and naphthalene, respectively in 3 days, 72.3% of 1,000 mg/L diesel oil in 7 days and 78.0% of 10,000 mg/L diesel oil in oil-contaminated soil during 28 days. Isolated strains Bacillus amyloliquefaciens S10 and B. subtilis GO9 can produce biosurfactant and formed 6.34 and 2.5 cm diameter of clear zones, respectively in oil-spreading test. Surface tension of their culture supernatant reduced from 74.6 to 34.4 and 33.3 mN/m, respectively during incubation, and critical micelle concentrations of culture supernatants were 2.0 and 5.9%, respectively. Consortium of A. calcoaceticus SL1 and B. amyloliquefaciens S10 degraded 77.8% of 10,000 mg/L diesel oil in 3 days, which indicated more efficient oil degradation than that by A. calcoaceticus SL1 alone. If these bacteria were applied together as a consortium to oil-contaminated sites, they may show a high removal rate of petroleum hydrocarbons.

Effect of Cu-resistant Pseudomonas on growth and expression of stress-related genes of tomato plant under Cu stress (구리-오염 토양에서 토마토 식물의 생장과 스트레스-관련 유전자 발현에 미치는 구리-내성 Pseudomonas의 영향)

  • Kim, Min-Ju;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.257-264
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    • 2017
  • Pseudomonas veronii MS1 and P. migulae MS2 have several mechanisms of copper resistance and plant growth promoting capability, and also can alleviate abiotic stress in plant by hydrolysis of a precursor of stress ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC) by ACC deaminase. In 4-week pot test for tomato growth in soil contained 700 mg/kg Cu, inoculation of MS1 and MS2 significantly increased root and shoot lengths, wet weight and dry weight of tomato plants compared to those of uninoculated control. The inoculated tomato plants contained less amounts of proline that can protect plants from abiotic stress, and malondialdehyde, an oxidative stress marker than those of control. ACC synthase genes, ACS4 and ACS6, and ACC oxidase genes, ACO1 and ACO4, both involved in ethylene synthesis, were strongly expressed in Cu stressed tomato, whereas significantly reduced in tomato inoculated with MS1 and MS2. Also, a gene encoding a metal binding protein metallothionein, MT2 showed similar expression pattern with above genes. All these results indicated that these rhizobacteria could confer Cu resistance to tomato plant under Cu stress and allowed a lower level of Cu stress and growth promotion.

Antimicrobial activities of Bacillus subtilis DS660 and Paenibacillus polymyxa DS842 (Bacillus subtilis DS660과 Paenibacillus polymyxa DS842의 항균활성)

  • Lee, Da-Sol;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.354-361
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    • 2018
  • This study was carried out to isolate soil bacteria with antimicrobial activity and evaluate antimicrobial substances produced by isolated bacteria. Among many isolates Bacillus subtilis DS660 and Paenibacillus polymyxa DS842 showed high antimicrobial activities against 6 species of microbial residents on human skin and 3 species of pathogenic bacteria. DS660 and DS842 showed 15.3~26.8 and 11.3~27.5 mm of inhibition zone diameter, respectively on nutrient agar medium against most target bacteria and fungi. DS660 and DS842 produced $57{\pm}8$ and $170{\pm}15{\mu}mol/ml$ of siderophore, respectively as an antimicrobial substance. Analysis of ethyl acetate extract of culture supernatants of DS660 and DS842 suggested production of glycolipid biosurfactant which reduced surface tension of culture supernatant of DS660 and DS842 from 60.0 to 40.3 and 30.3 mN/m, respectively. DS660 and DS842 also showed $169.2{\pm}9.9$ and $357.2{\pm}13.7nmol/min/mg$ protein of ${\beta}-1,3$-glucanase activity, respectively, and hydrolyzed cell wall components of 3 bacterial species. These results suggest that B. subtilis DS660 and P. polymyxa DS842 may be utilized as an environment-friendly biocontrol agent against some skin microbes and pathogenic bacteria.

Antimicrobial activities of Burkholderia sp. strains and optimization of culture conditions (Burkholderia sp. OS17의 항균활성 증진을 위한 배양최적화)

  • Nam, Young Ho;Choi, Ahyoung;Hwang, Buyng Su;Chung, Eu Jin
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.428-435
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    • 2018
  • In this study, we isolated and identified bacteria from freshwater and soil collected from Osang reservoir, to screen antimicrobial bacteria against various pathogenic bacteria. 38 strains were isolated and assigned to the class Proteobacteria (22 strains), Actinobacteria (7 strains), Bacteroidets (6 strains), and Firmicutes (3 strains) based on 16S rRNA gene sequence analysis. Among them, strain OS17 showed a good growth inhibition against 5 methicillin-resistant Staphylococcus aureus subsp. aureus strains and Bacillus cereus, Bacillus subtilis, Filobasidium neoformans. As a result of the 16S rRNA gene sequence analysis, strain OS17 show the high similarity with Burkholderia ambifaria $AMMD^T$, B. diffusa $AM747629^T$, B. tettitorii $LK023503^T$ 99.8%, 99.7%, 99.6%, respectively. We investigated cell growth and antimicrobial activity according to commercial culture medium, temperature, pH for culture optimization of strain OS17. Optimal conditions for growth and antimicrobial activity in strain OS17 were found to be: YPD medium, $35^{\circ}C$ and pH 6.5. When the strain was cultured in LB, NB, TSB, R2A media at $20^{\circ}C$ and $25^{\circ}C$, the antimicrobial activity did not show. Culture filtrate of strain OS17 showed antimicrobial activity against 5 MRSA strains, Bacillus cereus, Bacillus subtilis, and Filobasidium neoformans with inhibition zones from 2 to 8 mm. Optimal reaction time was 48 h in YPD medium, 100 rpm and 0.3 vvm in 2 L-scale fed-batch fermentation process for antimicrobial activity. Culture optimization of strain OS17 can be improved on antimicrobial activity. Therefore, the antimicrobial activity of Burkholderia sp. OS17 had potential as antibiotics for pathogens including MRSA.

Mass Cultivation of Rhodococcus sp. 3-2, a Carbendazim-Degrading Microorganism, and Development of Microbial Agents (카벤다짐 분해 미생물인 Rhodococcus sp. 3-2의 대량 배양 및 미생물 제제 개발)

  • Jun-Kyung Park;Seonghun Im;Jeong Won Kim;Jung-Hwan Ji;Kong-Min Kim;Haeseong Park;Yeong-Seok Yoon;Hang-Yeon Weon;Gui Hwan Han
    • Korean Journal of Environmental Agriculture
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    • v.42 no.4
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    • pp.259-268
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    • 2023
  • Rhodococcus sp. 3-2 strain has been reported to degrade benzimidazole-based pesticides, such as benomyl and carbendazim. Therefore, this study aimed to optimize culture medium composition and culture conditions to achieve cost-effective and efficient large-scale production of the Rhodococcus sp. 3-2 strain. The study identified that the optimal media composition for mass culture comprised 0.5% glucose, 0.5% yeast extract, 0.15% NaCl, 0.5% K2HPO4, 0.5% sodium succinate, and 0.1% MgSO4. Additionally, a microbial agent was developed using a 1.5-ton fermenter, with skim milk (20%), monosodium glutamate (15%), and vitamin C (2%) as key components. The storage stability of the microbial agent has been confirmed, with advantages of low temperature conservation, which helps to sustain efficacy for at least six months. We also assessed the benomyl degradation activity of the microbial agent within field soil. The results revealed an over 90% degradation rate when the concentration of viable cells exceeded 2.65 × 106 CFU/g after a minimum of five weeks had elapsed. Based on these findings, Rhodococcus sp. 3-2 strain can be considered a cost-effective microbial agent with diverse agricultural applications.

Antifungal Activity of Bacillus sp. AM-651 Against Phytophthora capsici (고추역병 유발병원균 Phytophthora capsici에 대한 Bacillus sp. AM-651의 항진균활성)

  • Lee, Jung-Bok;Shin, Jeong-Hak;Jang, Jong-Ok;Shin, Kee-Sun;Choi, Chung-Sik;Kim, Kun-Woo;Jo, Min-Sub;Jeon, Chun-Pyo;Kim, Yun-Hoi;Kwon, Gi-Seok
    • Microbiology and Biotechnology Letters
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    • v.36 no.3
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    • pp.227-232
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    • 2008
  • Biological antagonists of Phytophthora capsici were isolated from soil in Gyeongbuk, Korea. Among the isolated bacteria, a Bacillus sp. was identified from l6S rDNA sequence analysis and named Bacillus sp. AM-651. Bacillus sp. AM-65l strain which can strongly a antifungal activity against Phytophthora capsici. Culture conditions for the maximum production of the antagonistic substance were optimized. The production of antibiotic were high on modified Davis mineral medium pH 7 at $30^{\circ}C$. The medium for highest production of the agonistic substance optimized. It is composed the best activity on glucose, $(NH_4)_2SO_4$ and $K_2HPO_4$ at 0.5%, 0.1%, and 0.7%, respectively. By time course of culture solution selected Bacillus sp. AM-65l, the culture solution after 48hrs had strongly growth inhibition rate against P. capsici. And culture solution of Bacillus sp. AM-651 was stable within a pH range $5{\sim}11$ and temperature range $4{\sim}70^{\circ}C$. Bacillus sp. AM-651 cultured broth shown fungal growth inhibitory activity against B. sorokiniana, B. cinerea, R. solani avove and beyond P. capsici and comparatively showed a high activity against C. gloeosporioides, B. dothidea, B. cinerea and F. graminearum by agar diffusion method.

Cultivation Condition of Transformant Alcaligenes eutrophus Harboring Cloned phbC Gene for Production of P(3-hydroxybutyrate-3-hydroxyvalernte) Containing High Molar Fraction of 3-Hydroxyvalerate. (P(3-hydroxybutyrate-3-hydroxyvalerate)의 생산을 위한 재조합 phbC 유전자를 형질전환시킨 Alcaligenes eutrophus의 배양조건 검토)

  • 권순일;정영미;이용현
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.537-544
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    • 1998
  • The cultivation conditions of transformant Alcaligenes eutrophus AER5 harboring cloned phbC gene for mass production of poly (3-hydroxybutyrate-3-hydroxyvalerate)[P(3HB-3HV)] containing high molar fraction of 3-hydroxyvalerate (3-HV) were investigated. In two-stage batch cultivation, transformant accumulated P(3HB-3HV) containing 52.2 mol% of 3HV compared to 30 mol% of parent strain A. eutrophus H16. The increased 3-HV molar fraction was due to the amplified activity of PHB synthase participating in condensation of 3-HB and 3-HV. To increase efficiency of P(3HB-3HV) accumulation, fructose was added along with precursor compound valerate, and total cell mass and P(3HB-3HV) concentrations remarkably increased, but not 3-HV molar fraction. The effect of magnesium ion showed that P(3HB-3HV) concentration and 3-HV molar fraction were significantly increased upto 6.1 g/L and 71.3 mol% at 0.01 g/L of MgSO$_4$, respectively. The efficiency of several pH adjuster, NaOH, NaOH and (NH$_4$)$_2$SO$_4$, and NH$_4$OH, on total cell mass, p(3HB-3HV) concentration, and 3-HV molar fraction was also compared. To overcome the disadvantage of two-stage cultivation, one-stage intermittent fed-batch cultivation was attempted, such a way 10.0 g/L of fructose was supplied for cell growth at initial 36 hr and then 10.0 g/L of valerate and 5.0 g/L of fructose were applied to induce the accumulation of P(3HB-3HV), consequently, 10.4 g/L of P(3HB-3HV) with 38 mol% of 3-HV fraction could be obtained after 72 hr. These results can be used for elucidating cultivation strategy for mass production of P(3HB-3HV) containing high 3-HV molar fraction using transformant A. eutrophus AER5 harboring cloned phbC gene.

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Inhibitory Substance on the Snake Venoms Produced by Penicillium sp. (사독의 조해물질에 관한 연구)

  • Seu, Jung-Hwn;Yi, Dong-Heui
    • Microbiology and Biotechnology Letters
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    • v.7 no.2
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    • pp.75-89
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    • 1979
  • One strain of Penicillium sp. (175-66-B), isolated from soil, was able to produce a substance that has a strong inibition activity against the Agkistrodon and Trimeresurus venoms. In this experiment, the chemical and biological properties of the sample were investigated. As an inhibitory substance, it was effective to the proteinase, hemorrhagic and lethal factors of Agkistrodon and Trimeresurus venoms, and also effective to several fractions of the proteinases and hemorrhagic factors of Agkistrodon halys blomhoffi venom. Moreover, in the addition of prednisotone, it was more effective for the cure of the mouse envenomated with the venom amount of two fold of MLD$_{100}$. This substance was very stable to the acid, alkali and heat. Its melting point was high enough to sublime at 222$^{\circ}C$ without any decomposition. This sample was easily dissolved only in hot water, but not in several organic solvents except for a little dissolution in elate. It did not have the chelating activity. It had very strong specificity to the snake venoms. but its activity was depressed by the addition of zinc or cupric salts. This sample had no acute toxicity to the mouse. Its chemical formula was $C_{16}$ $H_{12}$$N_2$ $O_{10}$ with the molecular weight of about 392. It has two epoxy groups and four carboxyl radicals, but amino, nitrite and nitrate radicals, unsaturated bonds and aromatic ring were not detected. Theuchemical configuration of this sample was suggested to be;

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Changes of Microbial Activity and Physicochemical Environment during Composting of Papermill Sludge in a Pilot Plant (제지슬럿지의 퇴비화 과정 중 미생물활성 및 이화학적 환경변화)

  • Chung, Young-Ryun;Chung, Man-Hoon;Han, Shin-Ho;Oh, Say-Kyun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.3 no.2
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    • pp.79-89
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    • 1995
  • Changes of microbial activity and physicochemical environment during composting of papermill sludge(PMS) in the pilot plant equipped with an agitated bed reactor were monitored for establishing the efficient composting system. Microbial activity determined as the evolution of $CO_2$ increased for the first 10 days after introduction of PMS to the reactor and decreased thereafter. Population changes of microorganisms in the reactor-PMS were not typical as in windrow system. The ratio of thermophilic bacteria to mesophilic bacteria, however, increased slowly even 23 days after introduction. Temperature of PMS increased rapidly from the first day and reached $62^{\circ}C$ at 7 days after introduction and decreased slowly thereafter. The acidity of PMS was pH 6.8 initially, increased to pH 8.0 after 7 days and decreased to pH 7.4 after 23 days. Redox potential(Eh) of PMS was -320mV at the beginning of composting, but it was increased with time to reach -15mV after 23 days composting. However, Eh of PMS pre-sterilized before measurement was average 50mV, regardless of composting periods indicating the major role of microorganisms during composting process. Water content of PMS was 67% initially and decreased to about 50% after 23 days composting in the reactor. Less than 13 days-old compost inhibited growth of radish in the container mixture with bed soil. Based on statistical analysis of microbial and physicochemical parameters of PMS during composting, an equation was developed for determining compost maturity. A number of experiments using various organic wastes are required before application of the formular to the practical use.

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Studies on Identification and Drug Resistance of Atypical Mycobacteria isolated from Patients with Pulmonary Tuberculosis (폐결핵환자에서 비정형항산균의 분리, 동정 및 약제감수성에 관하여)

  • Chung, Dong-Hyun;Kim, Sung-Kwang;Kim, Joo-Deuk
    • Journal of Yeungnam Medical Science
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    • v.1 no.1
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    • pp.49-58
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    • 1984
  • The differential diagnosis of atypical mycobacteriosis caused by atypical mycobacteria (with the exception of Mycobacterium tuberculosis, Mycobacterium bovis, and Mycobacterium leprae) which are widly distributed in soil and water, from pulmonary tuberculosis is possible only when atypical mycobacteria are isolated and identified. In this investigation, attempts were made to isolate atypical mycobacteria from persons registered as tuberculosis patients in the Anyang Health Center in Anyang City, Kyungki province, Korea. Biological and biochemical tests were performed for the atypical mycobacteria isolated from these patients, also retrospective analysis of clinical and X-ray findings of the patients with bacteriologically confirmed atypical mycobacteriosis were done. The results can be summarized as follows: 1. 103 strains of mycobacteria were isolated among 334 sputum samples from patients. 2. Among the isolated mycobacteria, 10 strains (9.7%) were found to be a atypical mycobacteria and 93 strains (90.3%) were tubercle bacilli of human type. 3. On the basis of Runyon's grouping of atypical mycobacteria, there were 3 strains (30.0 %) of scotochromogen and nonphotochromogen respectively, 4 strains (40.0%) of rapid grower, and no photochromogen. 4. By biochemical tests, 3 strains of scotochromogen were identified as Mycobacterium scrofulaceum (2 strains) and Mycobacterium szulgai (1 strain) 3 strains of nonphotochromogen were Mycobacterium avium-complex (2 strains) and Mycobacterium terriae (1 strain), and 4 strains of rapid grower were Mycobacterium fortuitum (3 strains) and Mycobacterium chelonei. 5. In drug sensitivity tests, all 10 strains isolated atypical mycobacteria showed resistance to various concentration of INH and SM and low concentration (10mcg, 40mcg and 50mcg) of EB, TH, and CS, and were sensitive to only high concentration (20mcg and 100mcg) of EB, TH, CS, and RFP. 6. In analysis of clical findings by the patients with bacteriologically confirmed atypical mycobacteriosis, it was found that clinical symptoms of these patients appeared not to be mild than those of patients with pulmonary tuberculosis. The patients with atypical mycobacteriosis had been treated for pulmonary tuberculosis for a long time and they showed no improvement.

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