• Title/Summary/Keyword: Condition of isolation

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Detection of DNA from Dermatophytes by Polymerase Chain Reaction (Polymerase chain reaction에 의한 동물 유래 피부사상균 DNA의 검출)

  • Kim, Young-Wook;Yeo, Sang-Geon;Choi, Woo-Pil
    • Korean Journal of Veterinary Research
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    • v.42 no.3
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    • pp.363-370
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    • 2002
  • For the development of diagnostic polymerase chain reaction (PCR) to fungal infection by dermatophytes Trichophyton and Microsporum, detection of the fungal DNA by PCR and analysis of the DNA pattern were undertaken in the present study. A total of 15 strains were tested and those consisted of 3 reference strains and 12 isolates such as: reference strains of T mentagrophytes (downy type, ATCC 9533), T rubrum (IFO 6204) and M gypseum (ATCC 9083), and each isolate of T mentogrophytes (powdery type), T mentagrophytes (granular type), T mentogrophytes (purple-red type), T rubrum, T raubitschekii, T tonsurans, T equinum, T ajelloi, T verrucosum, M cookei, M nanum and M gypseum. The DNA were purely isolated from all strains of Trichophyton spp. and Microsporum spp. by a simple method partly consisted of disruption of fungal cells by lyophilization and grinding and extraction of fungal DNA without phenol treatment which is a routine procedure in DNA isolation. For the detection of fungal DNAs, optimal condition of PCR was determined as preheating once at $94^{\circ}C$ for 5 min, 35 cycles of denaturation at $94^{\circ}C$ for 1 min, annealing at $38^{\circ}C$ for 1 min and polymerization at $72^{\circ}C$ for 2 min, and 1 cycle of final extension at $72^{\circ}C$ for 5 min. In PCR using arbitrary primers AP-1 (5' ACCCGACCTG3') and AP-2 (5' ACGGGCCAGT3'), DNAs in various numbers and sizes were detected from different species of Trichophyton and Microsporum, while DNAs in similar size were also detected in all strains of Trichophyton spp. and Microsporum spp. There were unique DNAs observed from certain dermatophytes by AP-1 such as 1,900 bases in T rubrum, 950 and 1,100 bases in T raubitscheldi, 2,100 bases in T equinum, 400 bases in T verrucosum and 1,150 bases in M gypseum. The unique DNAs were also observed by AP-2 such as 1,200 bases in T ajelloi, 250 bases in T verrucosum, 1,150 bases in M cookei and 2,000 bases in M nanum. The results indicated that PCR can detect a specific DNA from certain Trychophyton and Microsporum spp, which can be the information for further development of diagoomc PCR to dennatophytes.

Isolation and Identification of Hydrolytic Enzyme-producing Bacteria from Spent Mushroom Substrate (버섯부산물유래 가수분해효소분비 박테리아의 분리 및 동정)

  • Kim, Young-Il;Jeong, Se-Hyung;Seok, Joon-Sang;Yang, Si-Yong;Huh, Jung-Won;Kwak, Wan-Sup
    • Journal of Animal Science and Technology
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    • v.50 no.5
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    • pp.713-720
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    • 2008
  • This study was conducted to isolate and identify xylanase- and cellulase-producing thermophilic bacteria from stacked spent mushroom substrates and to determine the optimal medium conditions for their growth. Bacteria with the highest xylanase and CMCase activities were strain 3 and 201-7. Both of them were identified as Bacillus spp. and named Bacillus subtilis KU3 and Bacillus subtilis KU201-7. The optimal medium condition of Bacillus subtilis KU3 was obtained when 3%(w/v) of yeast extract and 1%(w/v) of maltose were used as nitrogen and carbon sources, respectively. That of Bacillus subtilis KU201-7 was obtained when 0.5%(w/v) of yeast extract and 0.5%(w/v) of CMC were used as nitrogen and carbon sources, respectively.

Isolation of Phytase Producing Pseudomonas fragi and Optimization of its Phytase Production (Acid Phytase를 생산하는 Pseudomonas fragi의 분리와 phytase의 생산조건)

  • Kim, Young-Jin;Jang, Eun-Seok;In, Man-Jin;Oh, Nam-Soon
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.291-298
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    • 2003
  • A bacterial strain producing a high level of an extracellular phytase was isolated from livestock waste water, identified as a strain of Pseudomonas fragi and designated as Pseudomonas fragi Y9451. Under the phytase production medium, the activity of phytase reached the highest level after 120 hours of incubation. On the effect of carbon sources on the phytase production, the most favorable carbon source for phytase production was fructose. As for the effect of nitrogen sources, high levels of phytase activity were detected in the medium containing nutrient broth as the nitrogen source. Free $PO_4^{3-}$ inhibited phytase production with increasing concentration of $KE_2PO_4$ and phytate in the media. The addition of $CaCl_2$ and $MgSO_4$ also resulted in the inhibition of phytase production. To investigate the effect of aeration on the phytase production, different volumes of culture broth in Erlenmeyer flasks were incubated in rotary shaker at the speed of 200 rpm. As a result, a high level of phytase activity was detected at small volume of culture broth as compared to larger volume because of its more aerobic condition.

Studies on the scouring of raw silk by the application of bacterial enzyme of sericinase -(Part 1.) Isolation and selection of proper bacteria and some studies of properties of this bacterial enzyme- (세균성(細菌性) Sericin 분해(分解) 효소(酵素)에 의(依)한 생사정련(生絲精練)에 관(關)한 연구(硏究) -(제1보(第1報)) 유용세균(有用細菌)의 분리(分離) 세정(選定) 및 생성(生成) Sericinase 의 효소학적(酵素學的) 성질(性質)에 대(對)하여-)

  • Ki, Woo-Kyung;Seu, Jung-Hwn
    • Applied Biological Chemistry
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    • v.12
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    • pp.75-81
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    • 1969
  • In order to obtain a method of scouring of raw silk and degumming of cocoon by the appling of bacterial enzyme of sericinase, a strain of proper bacteria was isolated and some properties of the enzyme produced by the isolated bacteria were studied and the following results were obtained. 1) Optimum pH and temperature were indicating 7.5 and $50^{\circ}C$. and on these conditions, the silk fibroin will get no modification at all. 2) Sericinase activity was inhibited by calcium ion in the free incubation but same ions reacted as an activator in the reaction with substrate. So, degumming of Tussah cocoon which contains calcium oxalate in the cocoon layer will be possible by the treatment of this enzyme. 3) This bacterial sericinase never gives any affection to the fibroin layer of the silk. 4) The factors of smooth-surface condition, damage of fibroin layer, touching, luster and degumming effect of the silk resulting by the enzyme treatment were more appropriate than the results of other scouring methods, saponin, alkali and saponin-alkali using methods. We expect to get the same scouring result that compared with the papain or pancreatin were applied.

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Probiotic Characterization of Acid- and Bile-tolerant Lactobacillus salivarius subsp. salivarius from Korean Faeces

  • Bae, H.C.;Nam, M.S.;Lee, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1798-1807
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    • 2002
  • This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

Isolation and Identification of High Cellulolytic Bacteria from Spent Mushroom Substrate and Determination of Optimal Medium Conditions for the Growth (버섯폐배지로부터 섬유소분해력이 높은 중온성 균의 분리 및 균주생산을 위한 배지조건의 최적화)

  • Kim, Young-Il;Jung, Se-Hyung;Seok, Joon-San;Yang, Si-Yong;Huh, Jeong-Weon;Kwak, Wan-Sup
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.255-260
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    • 2007
  • This study was conducted to isolate and identify bacteria producing xylanase and cellulase from spent mushroom substrates and to determine the optimal medium conditions for their growth. Bacteria showing high xylanase and carboxymethyl cellulase activities and low protease and amylase activities were strain 201-3 and strain 206-3. Strain 201-3 was identified as Enterobacter ludwigii and named Ent. ludwigii KU201-3. 206-3 was identified as Bacillus cereus and named B. cereus KU206-3. The optimal medium condition of Ent. ludwigii KU201-3 was obtained when 1%(w/v) of soybean meal and 3%(w/v) of sucrose were used as nitrogen and carbon source, respectively. That of B. cereus KU206-3 was obtained when 3%(w/v) of soybean meal and 1%(w/v) of molasses were used as nitrogen and carbon sources, respectively.

Molecular Isolation and Characterization of the 2CysPrx Gene from Salicornia herbacea (퉁퉁마디로부터 2CysPrx 유전자 분리 및 특성 분석)

  • Kim, Suk-Kyu;Chung, Sang Ok;Na, Gil-Jong
    • Korean Journal of Environment and Ecology
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    • v.30 no.5
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    • pp.810-820
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    • 2016
  • This study is focused on the investigation of the genes which are induced by various stresses of the halophyte Salicornia herbacea. One of the factors influencing in the germination of Salicornia herbacea is salt stress. The highest germination rate was found in the condition without NaCl, and the upper limit of the NaCl concentration for the germination of Salicornia herbacea was 7%. The optimal temperature of $20^{\circ}C$showed a germination rate of 98%. Among genes induced by stress the 2CysPrx gene was cloned and analyzed for this study. The 2CysPrx gene has two cysteine conserved residues and is composed of 275 amino acids with molecular weight of 30.1kDa. The 2CysPrx gene appeared to be one copy in the genome and consists of 6 introns and 7 exons. Quantitative real-time PCR revealed that the highest transcription rate induced by NaCl and $H_2O_2$ appeared to be at the concentration of 3.5% NaCl and 40mM $H_2O_2$, respectively. The amount of transcript induced by high temperature($40^{\circ}C$) and $75{\mu}M$ of ABA was respectively highest. The gene at low temperature ($4^{\circ}C$) appeared not to be expressed. We are conducting to clone other peroxyredoxin genes induced by various environmental stresses.

Inhibitory Conditions of Asexual Development and their Application for the Screening of Mutants Defective in Sexual Development (Aspergillus nidulans에 있어서 무성분화(無成分化)의 억제조건(抑制條件)과 이를 이용(利用)한 유성분화결손(有性分化缺損) 돌연변이주(突然變異株)의 대량분리(大量分離))

  • Han, Dong-Min;Han, Yoo-Jeoung;Lee, Young-Hoon;Jahng, Kwang-Yeop;Jahng, Seung-Hwan;Chae, Keon-Sang
    • The Korean Journal of Mycology
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    • v.18 no.4
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    • pp.225-232
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    • 1990
  • In order to find an useful condition under which the mutants defective in sexual development could be isolated, the effects of several cultural conditions on the developments of Aspergillus nidulans were examined. Among several conditions found to restrict the asexual sporulation but enhance the sexual process, the interference of aeration by sealing the plates with sealing film was the most useful one for the purpose of mutant isolation. Sealing at any time before 20 hours from inoculation prevented both sexual and asexual process. When the seal was removed after 24 hours, however, the mycelia developed only to sexual organs. Using this properity, the early morphogenic process of sexual development could be easily observed and a number of mutants that showed some defects in the process could be isolated. The mutants were divided into 3 groups, NSD (never in sexual development), BSD (block in sexual development) and ASD (abnormal in sexual development). NSD mutants never produced either the $H{\ddot{u}}lle$ cells or cleistothecia and some produced the asexual organs even when the aeration was restricted. BSD mutants were blocked in the process of $H{\ddot{u}}lle$ cell, cleistothecia, crozier, asci or ascospore formation. ASD mutants had defects in the amount of cleistothecia, time of cleistothecial maturation or color of ascospores.

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Isolation of an Acetic Acid Bacterium Acetobacter pasteurianus CK-1 and Its Fermentation Characteristics (초산균 Acetobacter pasteurianus CK-1의 분리 및 발효 특성)

  • Bang, Kyu-Ho;Kim, Chae-Won;Kim, Chul-Ho
    • Journal of Life Science
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    • v.32 no.2
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    • pp.119-124
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    • 2022
  • To effectively isolate acetic acid bacteria for producing makgeolli vinegar, various products were researched, and Acetobacter pasteurianus CK-1, a strain that is excellent in acetic acid production, was finally isolated. The optimal growth temperature of the isolated strain was confirmed to be 30℃, and it grew well in the pH range of 5.5~6.5, with optimal growth at pH 6. A. pasteurianus CK-1 had the most active proliferation when the initial ethanol concentration in the medium was 4%, and growth was possible even at an ethanol concentration of 7%. When inoculating the isolated strain into makgeolli to induce acetic acid fermentation, the pH at the beginning of fermentation was 3.54, which was gradually lowered to 2.77 after 18 days of fermentation. The acidity was 0.75% at the beginning of fermentation and gradually started to increase from the 4th day of fermentation. The final acidity at the end of fermentation was 5.54%. In the vinegar fermented by inoculating A. pasteurianus CK-1, acetic acid content was detected to be as high as 3,575.7±48.6 mg%, and the malic acid and citric acid contents were 2,150.8±27.6 and 55.8±3.7 mg%, respectively. Further, it was confirmed that the content and ratio of the organic acids produced significantly differed depending on the type of inoculated bacterial strain. During acetic acid fermentation, the populations of yeast and A. pasteurianus CK-1 were inversely changed. In the initial stage of fermentation, yeast dominated, and after 10 days of fermentation, A. pasteurianus CK-1 slowly proliferated and reached stationary phase.

Antifungal Activity of Bacterial Strains isolated from Tidal Mudflat and Salted Seafood (traditional Jeotgal) Against Six Major Plant Pathogens (갯벌 및 젓갈에서 분리한 세균의 작물 주요 병원균에 대한 항균활성 효과 검정)

  • Kim, Tack-Soo;Lee, Ga-Hyung;Kim, Gyun-Jang;Lee, Se-Won;Park, Kyung-Seok;Park, Jin-Woo
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.421-426
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    • 2010
  • Many bacterial strains inhabit strong saline condition, such as tidal mudflat and salted seafoods, were identified and reported for the proposed protease activities and salt resistance; however antifungal activities against plant fungal pathogen have not well been studied until now. In this study, primary screening was performed for the isolation of promising strains against major plant pathogens like Sclerotinia sclerotiorum, Fusarium oxysporum, Phytophthora capsici, Botrytis cineria, Collectotrichum acutatum and Pythium ultimum. Totally 423 bacterial strain were isolated from laboratory media which was based on different morphological characteristics and all the strains were dual cultured against major fungal pathogens on PDA, finally 40 strains were selected as antifungal bacterial strain and identified by fatty acid phylogenic difference analysis from MIDI shorlock gas chromatography system. As a result, antifungal strains from tidal mudflat were 10 species of 6 genus. Paenibacillus macerans was dominant species; 5 strains among the 17 isolates from tidal mudflat. Antifungal strains from salted seafoods were 7 species of 3 genus and Bacillus atrophaeus was dominant species; 12 strains among the 23 isolates from salted fishes.