• Title/Summary/Keyword: High isolation

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Isolation and Characterization of Cryptococcus sp. CS-2 Secreting Polygalacturonase from Soil (토양으로부터 Cryptococcus sp. CS-2의 분리 및 균주가 분비하는 Polygalacturonase의 특성에 관한 연구)

  • 강희경;문명님;임채영;양영기
    • Korean Journal of Microbiology
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    • v.35 no.2
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    • pp.158-163
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    • 1999
  • A ploygalacturonase-produchg yeast was isolated from Cheju soil by selective eivichment media. One strain which has the highesl activity of polygalacturonase was selected. The characle~ishcs of the strain CS-2 were as follows: CS-2 utilized xylose. sucrose, maltose, u.ehalose, cellobiose. melibiose, lactose, raffinose, inosiiol, dulicilol, and dextrose, but did not utilized galactose, nitrate. nit~te, and lysine. Growth of CS-2 was inhibited by cyclohexamide, 1% acetic acid, and high concenaation (over 50%) of glucose. It grew at $30^{\circ}C$ but did 'IIOL $35^{\circ}C$. The cell size ofthe strain CS-2 was 2.9 p ~ n in length and 1.3 $\mu$ in diameter. Vegetable reproductmn was multiple budding and ascospre was present I to 4. Pseudomycelia or true myceliua formation were not observed In any of the cullureq. These results suggest that strain CS-2 is most likely a strain related Cryptococcus spp. (Cryptococcu spp. CS-2). When polygalacturonase or ihe yeast was induced by addition of polygalactoronic acid, polygalacturonase activity was detected in culture supernatent. There was a peak of specific activity a1 he mid-stationary phase(3 days culture) of growth. Polygalacturonase specific activity of Crylmcoccus sp. CS-2 was 2.96 unitsling. The molecular weighl ol'polygalacturonase was showed to be 46 KDa by both SDS-PAGE and activity stailling.

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Characterization and phylogenetic analysis of halophilic bacteria isolated from rhizosphere soils of coastal plants in Dokdo islands (독도 해안식물로부터 분리된 호염성 세균들의 특성 및 계통학적 분석)

  • You, Young-Hyun;Park, Jong Myong;Lee, Myung-Chul;Kim, Jong-Guk
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.86-95
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    • 2015
  • To study the halobacterial diversity at the rhizospheric soil of coastal plant native to Dokdo islands, several host plant were selected and its rhizospheric soil was sampled. Soil sample was diluted serially and pure isolation was done by sub-culture using marine agar media. 26 halophilic strains cultivable at the marine medium containig concentration of 9.0% sodium chloride were selected among total 161 isolates. Their partial 16S rRNA gene sequences extracted from genomic DNA were analyzed and partially identified. Furthermore, to identify their genetic relationship, phylogenetic tree was deduced. Total 26 strains were belongs to Firmicutes (30.8%), Gamma proteobacteria (53.8%), Bacteroidetes (7.7%), Alpha proteobacteria (7.7%), and Actinobacteria (7.7%). These results showed the specific difference from previous researches which has been reported the microbial flora of soil or sea water around the Dokdo islands. Furthermore, 4 among 26 halophilic strains grew at above 12.0% NaCl concentrated marine broth, and 2 strains Idiomarina abyssalis LM4H23 and Halomonas huangheensis AS4H13 grew at 15.0% concentration. These halophilic strains thought to overcoming the severe stress like high salt concentration or variation derived from Dokdo-specific climate and might have unknown, specific relationship with their host coastal plant native to Dokdo islands.

Isolation and identification of β-glucosidase producing halophilic Roseivivax roseus (β-Glucosidase를 생성하는 호염성 Roseivivax roseus 균주의 분리 및 분류동정)

  • Cho, Geon-Yeong;Han, Song-Ih
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.141-147
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    • 2015
  • Four halophilic bacteria were isolated from a salt water tank of more than 25% above salinity used for production of salt. HJS1 and HJS6 strains were identified as having ${\beta}$-glucosidase producing capabilities at high salinity. ${\beta}$-Glucosidase produced from these bacterial strains showed the best activity at 56-79 U/ml in NaCl (0-5%), showing the highest ${\beta}$-glucosidase activity at NaCl 3%. A salt tolerant ${\beta}$-glucosidase can maintain at least 75% activity of the enzyme in 0-20% NaCl concentration. The 16S rRNA gene sequences of strains HJS1 and HJS6 shows 99.8% similarity with Roseivivax roseus $BH87090^T$. Those sequences were registered as AB971835 and AB971836 in the NCBI GenBank. DNA-DNA hybridization test revealed that both strains showed 90.1 to 90.3% hybridization values with R. roseus $BH87090^T$, which was the closest phylogenetic neighbor. Major Cellular fatty acids of strains HJS1 and HJS6 were $C_{16:0}$, $C_{18:1}$ ${\omega}7c$, $C_{19:0}$ cyclo ${\omega}8c$ and 11-methyl $C_{18:1}$ and the major quinone was Q-10. Their fatty acid composition and quinone were very similar to Roseivivax roseus $BH87090^T$. Meanwhile, Roseivivax roseus $BH87090^T$ did not produce any ${\beta}$-glucosidase. Based on the molecular and chemotaxonomic properties, strains HJS1 and HJS6 were identified as members of Roseivivax roseus.

Regulation of Corynebacterium ammoniagenes purF and Isolation of purF-Specific Regulatory Proteins (Corynebacterium ammoniagenes에서 purF 유전자의 조절 및 이에 특이적인 조절 단백질의 분리)

  • Lee, Seok-Myung;Kim, Youn-Hee;Lee, Heung-Shick
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.233-238
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    • 2009
  • The expression of Corynebacterium ammoniagenes purF was analyzed by utilizing a plasmid carrying a cat gene fused to the purF promoter region. Adenine and guanine repressed the expression of the purF gene by 20~30% but hypoxanthine did not exert such repressive effect. The expression purF was maximal at the late log phase and remained constant throughout the stationary phase. Promoter $P_{180}$ which was developed in C. glutamicum was also functional in C. ammoniagenes, achieving maximal activity at the late log phase. The promoter outperformed Escherichia coli $P_{tac}$ promoter by 40~50% level. DNA-affinity purification identified a protein which could bind to the promoter region of the purF gene. The protein showed high similarity to the CRP-family transcriptional regulator encoded by NCgl0120 in C. glutamicum. The size of the screened protein agreed with the expected protein size from the ORF NCgl0120. The corresponding gene in C. ammoniagenes encoded a 42 kDa polypeptide composed of 400 amino acids with expected pI of 4.9. The encoded protein showed 14.1% and 15.8% identity with E. coli and Bacillus subtilis PurR, respectively, suggesting that the isolated protein might be a novel type of regulatory protein involved in the regulation of purine metabolism.

Microfungal flora of Tricholoma matsutake producing and nonproducing sites in the forest of Pinus densiflora (적송 (Pinus densiflora) 림내 송이(Tricholoma matsutake) 발생지와 미발생지의 토양 균류의 수직 분포)

  • Song, Hyun-Soon;Min, Kyung-Hee
    • The Korean Journal of Mycology
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    • v.19 no.2
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    • pp.109-119
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    • 1991
  • The vertical distribution of the fungal population for the soil samples from two sites of producing and nonproducing of Tricholoma matsutake, song-yi mushroom, were examined at Yang­yang and Myung-joo, Gangweon province. By the dilution plate method, a total number of propagu­les of fungi per gram of soil was observed to be low at the song-yi producing sites but high at the song-yi nonproducing sites under the communities of Pinus densiflora. The tendency of the number of fungal propagules were decreased with the increasing vertical depth. In the incuhation method at $42^{\circ}C$, six genera and nineteen species of the fungi were isolated from two sites; Aspergillus fumigatus, Acremonium sp., Talaromyces stipitatus, Penicillium lilacinum, P. oxalicum and Westerdykella multispora. The most dominant species by this method was A. fumigatus. From heat treatment method at $70^{\circ}C$, seven genera and nineteen species were isolated; Aspergillus fumigatus, Alternaria alternata, Neurospora sitophila and Mucor sp.. In the ethanol treatment method, one genera and one species was isolated Mortierella sp.. From the three isolation methods, it was found that the total number of the soil fungi and the frequency of species appeared were the highest at the soil of upper layer whereas the lowest at the soils of lower layer in its vertical distribution.

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Studies on Takjoo Yeasts (Part I) -Isolation and Identification of Takjoo Yeasts- (탁주효모(濁酒酵母)에 관(關)한 연구(硏究) (제1보(第一報)) -탁주효모(濁酒酵母)의 분리(分離) 및 동정(同定)에 대(對)하여-)

  • Park, Yoon-Joong;Lee, Suk-Kun;Oh, Man-Jin
    • Applied Biological Chemistry
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    • v.16 no.2
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    • pp.78-84
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    • 1973
  • The strains of 297 yeasts were isolated in TakJoo mashes of 12 breweries not using the cultivated yeast and then brewing test with each yeast were carried out. The strains of 7 yeasts that have high fermentative ability among the isolated strains were selected and identified. The results obtained were as follows: 1) In the brewing test with the isolated yeasts of each brewery, average alcohol percentage of each mash had a little differences as $13.20{\sim}15.20$ percentage. 2) In fermentative lest, the isolated yeasts from the first stage mash and from the second stage mash showed t little differences in the average alcohol percentage of mash. 3) The fermentative test using the isolated yeasts based on TTC stain had at little differences. 4) Among 7 strains selected, strains: Dm-1, Dm-2, Y-1, and T-1 appeared TTC pink yeast; strsins:C-1, C-2 and Gs-1 appeared TTC red one. 5) It was identified that strains: Dm-1, Y-1, C-1, C-2 and T-1 were Sac. cerevisae; the strain Gs-1 were Sac. pretoriencis; strain D-2 were Sac. rouxii.

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Improvement of Degrading Activity of Poly(butylene succinateco-butylene adipate)-Degrading Strains Isolated from Soils (토양에서 분리한 Poly(butylene succinate-co-butylene adipate) 분해균의 분해활성 증진)

  • Joo, Hyun-Jin;Kim, Mal-Nam
    • Korean Journal of Environmental Biology
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    • v.27 no.2
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    • pp.198-204
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    • 2009
  • From leaf mold and reclamation site soil of the Capital area of Korea, 3 poly(butylene succinate-co-butylene adipate: PBSA)-degrading strains were isolated through the clear zone test. The PBSA-degrading activities of the strains were assessed by means of a modified Sturm test using 0.01% of PBSA film as a sole carbon source. After the modified Sturm tests for 40 days at the respective isolation temperatures, the 3 strains degraded 30%, 55% and 43% of PBSA, respectively. The isolated strains were identified to be Burkholderia cepacia PBSA-4, Bacillus licheniformisPBSA-5 and Burkholderia sp. PBSA-6 through the 16S rDNA gene sequence analysis. Among them, PBSA-5 degraded both PBSA and Poly(vinyl alcohol). The degradation activity of the PBSA degrading strains appeared to be high at moderate temperatures such as $27^{\circ}C$ and $37^{\circ}C$, and initial inoculum size of $10^{10}cfu\;mL^{-1}$ degraded PBSA 1.2~1.3 more times than that $10^9cfu\;mL^{-1}$. Addition of 0.1 or 0.5% (w/w) of gelatin, yeast extract and ammonium sulfate raised the PBSA degrading activity, and especially addition of 0.1% (w/w) of gelatin enhanced the PBSA degrading activity by more than 33%. The mixed strains degraded PBSA faster than the single strain.

Isolation and Identifieation of Entomopathogenic Nematodes from Soil and Insect (토양과 곤충 사체로부터 곤충병원성 선충의 분리 및 동정)

  • 한상미;한명세
    • Korean Journal of Environmental Biology
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    • v.17 no.3
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    • pp.321-330
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    • 1999
  • Nematodes were isolated using silkwom trap through the investigation of 100 soil samples from various biotopes in Korea. The 30 nematode strains from soil and dead insects by the pathogenicity aganinst silkworms (Bombyx mori mori) and insect pests of Calliphora vomitoria, Pseufazetia separata, Palomena angulosa, and Melolontha incana. Mortailty of the silkworm larvae and pupae were as high as 100% by nematode infection, those of insect of pests were varied from 20 to 100%. The 30 strains of entemopathogenic nematodes were classified into five groups of Rhabditidae, Diplogatroidae, Heterorhabitidae, Steinernematidae, and Tylenchida by morphological criteria. The genetic relationships among the 30 nematode strains were analyzed by various RAPD bands with twenty primers. The 30 nematode strains were classified into six major subgroups on the basis of the genetic similarity coefficient of 0.853. The grouping by RAPD was agree with those of morphological taxa in discrimination of the higher group, however, was not completely agree in the subgroup. The family Steinernematidae belong to Rhabditida was clarified as closer to the Tylenchida, rather than the other Rhabditida of Heterorhabitidae, Rhabditidae, and Diplogatroidae in genetic distance valule. From the result of the morphological classification and RAPD of the genomic DNA showed that genetic relationship analysis furnish infurmation on phylogenetic classification and relationships of entomopathogenic nematodes. The application of genetic similarity will overcome the limitation of taxonomy and classification of morphologically simple nematode. Several primers were confirmed those utility of identification for individual nematode strains, the methods of molecular genetics secured the simplicity, rapidity and accuracy on the selection of entomopathogenic nematodes.

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Isolation and Characterization of Oligotrophic Strains with High Enzyme Activity from Buckwheat Sokseongjang (메밀 속성장 유래 효소활성 우수 저영양성 균주 분리 및 특성)

  • Lee, Sung-Young;Kim, Ji-Yeun;Baek, Sung-Yeol;Yeo, Soo-Hwan;Koo, Bon-Sung;Park, Hye-Young;Choi, Hye-Sun
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.735-741
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    • 2011
  • Bealmijang is a short-term fermented regional product that is prepared with soybean and extra ingredients. In this study, starter strain candidates were screened from Bealmijang for fermented soybean paste products. Twenty one bacterial strains producing extracellular enzymes (amylase, cellulase, protease, xylanase and lipase) were isolated from Bealmijang, buckwheat sokseongjang. The isolates were assessed for fibrinolytic and antibacterial activities, and salt tolerance. Strain HJ18-4, identified as Bacillus subtilis (AB601598) by biochemical properties (89.6%) and 16S rDNA sequencing (100%), showed the highest enzymatic, fibrinolytic, and antibacterial activities among the isolates. Although the growth of HJ18-4 was inhibited by the increase of NaCl concentration, the growth still exceeded that of B. subtilis KACC 10114 at 5% and 10% NaCl. These results suggest that B. subtilis HJ18-4 is suitable as a starter for soybean paste manufacture.

Isolation and Purification of Berberine in Cortex Phellodendri by Centrifugal Partition Chromatography (Centrifugal Partition Chromatography에 의한 황백으로부터 Berberine의 분리 및 정제)

  • Kim, Jung-Bae;Bang, Byung-Ho
    • The Korean Journal of Food And Nutrition
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    • v.27 no.3
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    • pp.532-537
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    • 2014
  • Cortex Phellodendri (CP) is derived from the dried bark of Phellodendron amurense. It has been widely used as a drug in traditional Korea medicine for treating diarrhea, jaundice, swelling pains in the knees and feet, urinary tract infections, and infections of the body surface. Many analytical methods have been used to study oriental herbal medicines, such as thin-layer chromatography, column liquid chromatography, and high performance liquid chromatography (HPLC). In this study, preparative centrifugal partition chromatography (CPC) was successfully carried out in order to separate pure compounds from a CP methanol extract. The optimum two-phase CPC solvent system was composed of n-butanol: acetic acid: water (4:1:5 v/v/v). The flow rate of the mobile phase was 3 mL/min in ascending mode with rotation at 1,000 rpm. The CPC-separated fraction and purification procedures were carried out by preparatory HPLC. The $^1H$ NMR spectrum revealed that the resonances at ${\delta}$ 4.10 and 4.20 ppm corresponded to three protons ($-OCH_3$), whereas those at ${\delta}$ 6.10 ppm corresponded to two protons ($-OCH_2O-$). Further, two aromatic protons (H-11 and H-12) conveys a doublet-doublet pattern. The H-11 doublet and H-12 doublet appear at ${\delta}$ 7.98 and 8.11, respectively. The $^{13}C$ NMR. spectrum showed a tetrasubstituted with a methylenedioxy group at C2 and C3, and two methoxy groups at C9 and C10. The chemical structure of the berberine was identified by $^1H$, $^{13}C$-nuclear magnetic resonance and electrospray ionization-mass spectroscopy spectral data analysis.