• Title/Summary/Keyword: strain characterization and identification

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Isolation and Identification of Yeasts from Wild Flowers Collected around Jangseong Lake in Jeollanam-do, Republic of Korea, and Characterization of the Unrecorded Yeast Bullera coprosmaensis

  • Han, Sang-Min;Hyun, Se-Hee;Lee, Hyang Burm;Lee, Hye Won;Kim, Ha-Kun;Lee, Jong-Soo
    • Mycobiology
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    • v.43 no.3
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    • pp.266-271
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    • 2015
  • Several types of yeasts were isolated from wild flowers around Jangseong Lake in Jeollanam-do, Republic of Korea and identified by comparing the nucleotide sequences of the PCR amplicons for the D1/D2 variable domain of the 26S ribosomal DNA using Basic Local Alignment Search Tool (BLAST) analysis. In total, 60 strains from 18 species were isolated, and Pseudozyma spp. (27 strains), which included Pseudozyma rugulosa (7 strains) and Pseudozyma aphidis (6 strains), was dominant species. Among the 60 strains, Bullera coprosmaensis JS00600 represented a newly recorded yeast strain in Korea, and its microbiological characteristics were investigated. The yeast cell has an oval-shaped morphology measuring $1.4{\times}1.7{\mu}m$ in size. Bullera coprosmaensis JS00600 is an asporous yeast that exhibits no pseudomycelium formation. It grew well in vitamin-free medium as well as in yeast extract-malt extract broth and yeast extract-peptone-dextrose (YPD) broth, and it is halotolerant growing in 10% NaCl-containing YPD broth.

Identification and characteristics of Lactic Acid Bacteria Isolated from Nuruk (누룩으로부터 젖산세균의 분리 및 특성)

  • 이정훈
    • KSBB Journal
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    • v.15 no.4
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    • pp.359-365
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    • 2000
  • Three lactic acid bacteria (C-1 K-3 and T-1 strain) were isolated from Nuruk and characterized subsequently. They were useful strains for production of lactic acid and their growth was inhibited at 10% ethanol pH 4 These strains were identified as lactococcus lactis subsp. lactis NR C-1 Leuconostoc mesenteroides subsp. mesenterides NR K-3 and pediococcus pentosaceus NR T-1 respectively by morphological physiological and biochemical characterization Lac lactis subsp lactis NR C-1 showed the highest lactic acid productivity. Leu measenteroides subsp mesenteroides NR K-3 showed stable lactic acid productivity and its growth was inhibited at pH 4. P pentosaceus NR T-1 had lower lactic acid productivity than the other two bacteria but it could not grow at 10% ethanol pH 4 The lactic acid productivity of these three strains in MRS broth were higher than that in Skim milk media the optimum pH and temperature for the lactic acid production of the three strains were 30-32$^{\circ}C$ and pH 6.0∼6.8 Glucose was the optimal carbon souorce for the lactic acid production. In terms of antagonism lac lactis subsp lactis NR c-1 showed somewhat inhibitory efects against some Gram positive rod and cocci such as Lactobacillus brevis and Streptococcus mitis. And Leu mesenteroides subsp mesenteroides NR K-3 showed the inhibitory effects against Streptococcus mitis but P. pentosaceus NR T-1 didn't show any inhibitory effects against tested strains.

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Characterization and Identification of Bacillus thuringiensis subsp. tenebrionis SR6 and SR8

  • Kim, Il-Gi;Lee, Jae-Wook;Suh, Suk-Chul;Rhim, Seong-Lyul
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.772-776
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    • 2004
  • Physiological and molecular characteristics of Bacillus thuringiensis SR6 and SR8 were investigated, and phase contrast and electron microscopies revealed that a large rhomboidal crystal protein was present in the sporulating cells. SDS-PAGE and Western blot analyses showed that B. thuringiensis SR8 produced 70 kDa protein much more than other proteins, and that the 70 kDa protein could bind to the antibody of B. thuringiensis subsp. tenebrionis-crystal toxin protein, indicating that the crystal 70 kDa protein has an immunological homology with B. thuringiensis subsp. tenebrionis-crystal toxin protein. The DNA fragment of B. thuringiensis subsp. tenebrionis-toxin gene was detected in B. thuringiensis SR6 and SR8 by using PCR amplification analysis. Furthermore, the insect bioassay showed the insecticidal activity against Colorado potato beetle larvae. Based on the physiological and molecular similarities to B. thuringiensis subsp. tenebrionis, it is suggested that the B. thuringiensis SR6 and SR8 may be mutants of the B. thuringiensis subsp. tenebrionis strain overexpressing the crystal of 70 kDa toxin protein.

Characterization of Tryptamine-Producing Bacteria Isolated from Commercial Salted and Fermented Sand Lance Ammodytes personatus Sauces (시판 까나리(Ammodytes personatus) 액젓에서 분리한 tryptamine 생성균의 특성)

  • Um, In-Seon;Kim, Tae-Ok;Kim, Hee-Dai;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.6
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    • pp.792-799
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    • 2016
  • We isolated seven tryptamine-producing bacteria from commercial salted and fermented sand lance (Ammodytes personatus) sauces using an L-tryptophan decarboxylating medium. These tryptamine-producing bacteria, identified using an API kit and 16S rRNA analysis, included Lysinibacillus xylanilyticus (one strain), Lysinibacillus fusiformis (four strains), and Staphylococcus epidermidis (two strains). Lysinibacillus spp. produced the highest levels of tryptamine in culture broth containing 0.5% L-tryptophan, compared with 1.0% and 2.0% preparations. After 72 h of incubation, Staphylococcus epidermidis produced the highest levels of tryptamine ($60.50{\mu}g/mL$ and $664.86{\mu}g/mL$) in culture broth containing 2.0% L-tryptophan. While Lysinibacillus spp. comprised the dominant tryptamine-producing bacteria in sand lance sauces, Staphylococcus epidermidis also showed high tryptamine-producing activity. This is the first report on the isolation and identification of tryptamine-producing bacteria in sand lance sauces.

Identification and Characterization of the Antifungal Substances of a Novel Streptomyces cavourensis NA4

  • Pan, Hua-Qi;Yu, Su-Ya;Song, Chun-Feng;Wang, Nan;Hua, Hui-Ming;Hu, Jiang-Chun;Wang, Shu-Jin
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.353-357
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    • 2015
  • A new actinomycete strain NA4 was isolated from a deep-sea sediment collected from the South China Sea and showed promising antifungal activities against soilborne fungal pathogens. It was identified as Streptomyces cavourensis by morphological, physiological, and phylogenetic analyses based on its 16S rRNA gene sequence. The main antifungal components were isolated and identified from the fermentation culture as bafilomycins B1 and C1. These compounds exhibited significant antifungal activities and a broad antifungal spectrum. The results suggest that the Streptomyces cavourensis NA4 and bafilomycins B1 and C1 could be used as potential biocontrol agents for soilborne fungal diseases of plants.

Isolation and Identification of Lactobacillus plantarum CIB 001 with Bile Salt Deconjugation Activity from Kimchi (김치로부터 담즙산 분해능이 우수한 Lactobacillus plantarum CIB 001의 분리 및 동정)

  • Cha, Sang-Do;Kim, Tae-Woon;Lee, Dong-Hee
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.222-226
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    • 2010
  • This study was carried out to isolate and characterize the Lactobacillus plantarum with bile salt deconjugation activity that was isolated from Kimchi. Some isolates were selected and identified as L. plantarum by 16S rRNA gene sequence and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of whole cell protein patterns. They were assayed to determine their capacities to express bile salt hydrolase (BSH) activity. Among the identified strains, L. plantarum CIB 001 showed the highest level of BSH activity. Then, resistance to gastric acidity and bile condition were analyzed for further characterization. This strain was able to maintain viability for 1h at pH 2.0 and to survive in a MRS (deMan, Rogosa, and Sharpe) broth with 1.0% of bile acids. L. plantarum CIB 001 would potentially be useful in the food industry as probiotics.

Cloning, Sequencing, and Characterization of Enterotoxin Pathogenicity Islet from Bacteroides fragilis 419

  • Rhie, Gi-Eun;Chung, Gyung-Tae;Lee, Yong-Jin;Sung, Won-Keun;Oh, Hee-Bok
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.86-90
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    • 2000
  • We have earlier reported on the cloning and identification of bft-k from an enterotoxigenic strain of Bacteroides fragilis 419, which was isolated from the blood of a Korean patient who suffered from systemic infections [4,5]. The bft-k gene encodes a 397-amino-acids metalloprotease enterotoxin, and the protein has been identified as a new isoform of B. fragilis enterotoxins (BFTs), which are cytopathic to intestinal epithelial cells to induce fluid secretion and tissue damage in ligated intestinal loops [4, 6, 18, 20]. This report describes the cloning and sequencing of the enterotoxin pahogenicity islet of B. fragilis 419 which contains the bft-k gene. the cloned enterotoxin pathogenicity islet was found to have 6,045 bp in length and to contain 120bp direct repeats near its end. In the pathogenicity islet, in addition to the BFR-K, two putative open reading frames (ORFs) were identified; (1) the t-3 gene encoding a 396-amino-acids protein of a putative metalloprotease; (2) the third gene encoding an ORF of a 59-amino-acids protein, whose function has not yet beenn characterized. The expression of the t-3 gene in B. fragilis 419 was verified by western blot analysis.

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Analysis of polymorphic region of GAM-1 gene in Plasmodium vivax Korean isolates

  • Kho, Weon-Gyu;Chung, Joon-Yong;Hwang, Ui-Wook;Chun, Jin-Ho;Park, Yeong-Hong;Chung, Woo-Chul
    • Parasites, Hosts and Diseases
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    • v.39 no.4
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    • pp.313-318
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    • 2001
  • The identification , characterization and quantification of Plasmodium sp. genetic polymorphism are becoming increasingly important in the vaccine development. We investigated polymorphism of Plasmodium vivax GAM-1 (PvGAM-1) gene in 30 Korean isolates. The polymorphic region of the PvGAM-1 gene, corresponding to nt 3792-4029, was amplified using polymerase chain reaction (PCR) followed by sequencing. All of the P. viuax Korean isolates were one type of GAM-1 gene, which were identical to that of the Belem strain. It is suggested that PvGAM-1 could not be used as a genetic marker for identifying or classifying P. vivax Korean isolates. It revealed that the polymorphic pattern as acquired basically by duplication and modification or deletion event of a 33 bp-motif fragment ended by poly guanine (G) and that there were at least three complete and one partial 33 Up-motif sequences within the polymorphic region in the longest cases such as those of South Korean and Belem isolates. In addition, we clustered P. vivax isolates with parsimonious criteria on the basis of PvGAM- 1 polymorphic patterns (insertion/deletion patterns) .

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Production and Characterization of a Novel Protease from Bacillus sp. RRM1 Under Solid State Fermentation

  • Rajkumar, Renganathan;Ranishree, Jayappriyan Kothilmozhian;Ramasamy, Rengasamy
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.627-636
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    • 2011
  • A commercially important alkaline protease, produced by Bacillus sp. RRM1 isolated from the red seaweed Kappaphycus alvarezii (Doty) Doty ex Silva, was first recognized and characterized in the present study. Identification of the isolated bacterium was done using both biochemical characterization as well as 16S rRNA gene sequencing. The bacterial strain, Bacillus sp. RRM1, produced a high level of protease using easily available, inexpensive agricultural residues solid-state fermentation (SSF). Among them, wheat bran was found to be the best substrate. Influences of process parameters such as moistening agents, moisture level, temperature, inoculum concentration, and co-carbon and co-nitrogen sources on the fermentation were also evaluated. Under optimized conditions, maximum protease production (i.e., 2081 U/g) was obtained from wheat bran, which is about 2-fold greater than the initial conditions. The protease enzyme was stable over a temperature range of 30-$60^{\circ}C$ and pH 6-12, with maximum activity at $50^{\circ}C$ and pH 9.0. Whereas the metal ions $Na^+$, $Ca^{2+}$, and $K^+$ enhanced the activity of the enzyme, others such as $Hg^{2+}$, $Cu^{2+}$, $Fe^{2+}$, $Co^{2+}$, and $Zn^{2+}$ had rendered negative effects. The activity of the enzyme was inhibited by EDTA and enhanced by $Cu^{2+}$ ions, thus indicating the nature of the enzyme as a metalloprotease. The enzyme showed extreme stability and activity even in the presence of detergents, surfactants, and organic solvents. Moreover, the present findings opened new vistas in the utilization of wheat bran, a cheap, abundantly available, and effective waste as a substrate for SSF.

Characterization of Dephosphocoenzyme A Kinase from Streptomyces peucetius ATCC27952, and Its Application for Doxorubicin Overproduction

  • Lee, Na-Rae;Rimal, Hemraj;Lee, Joo-Ho;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1238-1244
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    • 2014
  • Dephosphocoenzyme A (CoaE) catalyzes the last step in the biosynthesis of the cofactor coenzyme A. In this study, we report the identification and application of CoaE from Stretomyces peucetius ATCC27952. After expression of coaE, the protein was found to have a molecular mass of 28.6 kDa. Purification of the His-tagged fused CoaE protein was done by immobilized metal-affinity chromatography, and then in vitro enzymatic coupling assay was performed. The increasing NADH consumption with time shed light on the phosphorylating activity of CoaE. Furthermore, the overexpression of coaA and coaE independently under the $ermE^*$ promoter in the doxorubicin -producing wild type strain, resulted in 1.4- and 1.5-fold enhancements in doxorubicin production, respectively. In addition, the overexpression of both genes together showed a 2.1-fold increase in doxorubicin production. These results established a positive role for secondary metabolite production from Streptomyces peucetius.