• 제목/요약/키워드: Phenotypic characterization

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Characterization of Potato Scab Pathogens (Streptomyces Species) in Korea

  • Park, Duck-Hwan;Shrestha, Rosemary;Hur, Jang-Hyun;Lim, Chun-Keun
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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    • pp.162-165
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    • 2005
  • Potato scab, an important disease that affects developing tubers, causes a major problem in potato cultivation. The major potato cultivation areas in Korea are located in two Northern provinces, Gangwon and Gyeonggi, and two Southern provinces, Jeju island, and South Jeolla. In these areas, potato scab is widely distributed and has caused severe problem in potato cultivation. Therefore, potato-growing areas were surveyed for identification and distribution of potato scab pathogens from 1996 to 1999. Pathogenic Streptomyces strains were isolated from potato scab lesions and six representative Streptomyces species were characterized based on their phenotypic and molecular characteristics including, pathogenicity, physiological and morphological properties, analyses of 16SrRNA genes and 16S-23S ITS region, DNA relatedness, production of thaxtomin A, and the presence of nec1 and ORFtnp gene homologs. Three species were identified as previously described Streptomyces scabies, S. turgidiscabies, and S. acidiscabies, while other three species having distinct phenotypics properties were identified as novel S. luridiscabiei, S. puniciscabiei, and S. niveiscabiei.

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Production of Novel Cell-Associated Tannase from Newly Isolated Serratia ficaria DTC

  • Belur, Prasanna D.;Gopal, Mugeraya;Nirmala, K.R.;Basavaraj, N.
    • Journal of Microbiology and Biotechnology
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    • 제20권4호
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    • pp.732-736
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    • 2010
  • Five strains of tannic acid degrading bacteria were isolated and identified by phenotypic characterization. All the five isolates showed cell-associated activity, whereas only three showed extracellular activity. Serratia ficaria DTC, showing the highest cell-associated activity (0.29 U/l), was selected for further shake-flask studies. Tannase synthesis was growth associated and reached the peak in the late stationary phase of growth. Organic nitrogen sources enhanced the tannase production. Peak tannase production of 0.56 U/l was recorded in the medium having the initial pH of 6. The pH and temperature optima of the enzyme were found to be 8.9 and $35^{\circ}C$, respectively. This is the first report of cell-associated activity in the case of bacterial tannase. Cell-associated tannase of Serratia ficaria DTC could be industrially important from the perspective of its activity at broad temperature and pH ranges, and its unusually high activity at pH 8.9.

Flavobacterium amnigenum sp. nov. Isolated from a River

  • Patil, Kishor Sureshbhai;Padakandla, Shalem Raj;Chae, Jong-Chan
    • Journal of Microbiology and Biotechnology
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    • 제28권9호
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    • pp.1536-1541
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    • 2018
  • A yellowish, flexirubin-pigment-producing strain $I3-3^T$ was isolated from river water in Iksan, the Republic of Korea. The strain was gram-negative, aerobic, non-motile, showed catalase and oxidase activities, and could grow at a temperature range of $10-35^{\circ}C$, pH 5.0-10 and 0-2.0% (w/v) of NaCl. The major fatty acids were iso-$C_{15:0}$, iso-$C_{17:0}$ 3-OH and summed feature 3 (comprising $C_{16:1}{\omega}7c$ and/or $C_{16:1}{\omega}6c$). The isolate contained phosphatidylethanolamine, one aminolipid, and two unidentified lipids as the major polar lipids. Menaquinone-6 (MK6) was the major respiratory quinone. The G+C content of the genomic DNA of strain $I3-3^T$ was 35.6%. Comparison of the 16S rRNA gene sequence with the sequences of the closely related type strains showed highest sequence similarity of 96.95% and 96.93% to Flavobacterium nitrogenifigens $NXU-44^T$ and Flavobacterium compostarboris $15C3^T$, respectively. Based on phenotypic and phylogenetic distinctiveness, strain $I3-3^T$ is considered as a member of novel species within the genus Flavobacterium, for which Flavobacterium amnigenum sp. nov. is proposed. The type strain is $I3-3^T$ (=KCTC $52884^T$ =NBRC $112871^T$).

Medium optimization for keratinase production by a local Streptomyces sp. NRC 13S under solid state fermentation

  • Shata, Hoda Mohamed Abdel Halim;Farid, Mohamed Abdel Fattah
    • Journal of Applied Biological Chemistry
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    • 제56권2호
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    • pp.119-129
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    • 2013
  • Thirteen different Streptomyces isolates were evaluated for their ability to produce keratinase using chicken feather as a sole carbon and nitrogen sources under solid state fermentation (SSF). Streptomyces sp. NRC 13S produced the highest keratinase activity [1,792 U/g fermented substrate (fs)]. The phenotypic characterization and analysis of 16S rDNA sequencing of the isolate were studied. Optimization of SSF medium for keratinase production by the local isolate, Streptomyces sp. NRC13S, was carried out using the one-variable-at-a-time and the statistical approaches. In the first optimization step, the effect of incubation period, initial moisture content, initial pH value of the fermentation medium, and supplementation of some agro-industrial by-products on keratinase production were evaluated. The strain produced about 2,310 U/gfs when it grew on chicken feather with moisture content of 75% (w/w), feather: fodder yeast ratio of 70:30 (w/w), and initial pH 7 using phosphate buffer after 8 days. Based on these results, the Box-Behnken design and response surface methodology were applied to find out the optimal conditions for the enzyme production. The corresponding maximal production of keratinase was about 2,569.38 U/gfs.

Screening and Characterization of Pro biotic Lactic Acid Bacteria Isolated from Korean Fermented Foods

  • Lim, Sung-Mee;Im, Dong-Soon
    • Journal of Microbiology and Biotechnology
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    • 제19권2호
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    • pp.178-186
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    • 2009
  • To examine their potential as probiotics, acid and bile tolerance, antibiotics resistance, adhesion capacity to Caco-2 and HT-29, and antibacterial activity, of LAB isolated from Korean fermented foods such. as dongchimi, kimchi, Meju, and doenjang were assayed against foodborne pathogenic bacteria. DC 55, DC 136, DC 222, KC 21, KC 24, KC 34, KC 43, KC 117, MJ 54, MJ 301, SP 33, and SP 170 strains were resistant to acid and bile conditions. In particular, DC 55, DC 136, KC 24, KC 43, and MJ 301 strains were highly resistant to higher than 20 ${\mu}g/ml$ concentrations of vancomycin, streptomycin sulfate, or amoxicillin, whereas, DC 222, KC 21, KC 34, KC 117, MJ 54, and SP 33 strains were susceptible to lower than 2 ${\mu}g/ml$ concentrations of those antibiotics. The adhesion to HT-29 and Caco-2 cells varied with the strains tested in a strain-dependent manner. The highest level of adhesion was observed with DC 55, KC 21, KC 24, and MJ 301 strains, having higher than 50% of adhesion to HT-29 or Caco-2 cells. In addition, Staphylococcus aureus was the most sensitive to KC 21, showing an inhibition of about 70%, and the antibacterial activity of KC 21 against S. aureus resulted most likely from both organic acids and bacteriocin. Based on its phenotypic characteristics and utilization of various sugars, the KC 21 strain was identified as Lactobacillus plantarum.

난분해성 케라틴 폐기물 처리를 위한 우모 분해 미생물의 분리 및 특성 (Isolation and Characterization of Duck Feather-Degrading Microorganism for Treatment of Recalcitrant Keratinous Waste)

  • 고태훈;정진하;이나리;정성윤;박근태;손홍주
    • 한국환경과학회지
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    • 제21권2호
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    • pp.253-261
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    • 2012
  • We isolated and characterized novel duck feather-degrading bacteria producing keratinase. Twelve strains were isolated from soil and faces at poultry farm, and decayed feathers. They were identified as Bacillus methylotrophicus, Pseudomonas geniculata, Pseudomonas hibiscicola, Exiquobacterium profundum, Bacillus pumilus, Bacillus amyloliquefaciens, Chryseobacterium indologenes, Bacillus thuringiensis, Thermomonas koreensis, respectively, by phenotypic characters and 16S rRNA gene analysis. Generally, the level of keratinase production was not proportional to feather degradation rate. The highest keratinolytic activity was observed in the culture inoculated with Chryseobacterium indologenes D27. Although all strains did not degrade human hair, strains tested effectively degraded chicken feather(53.8-91.4%), wool(40.4-93.0%) and human nail (51.0-82.9%). These results suggest that strains isolated could be not only used to improve the nutritional value of recalcitrant feather waste but also is a potential candidate for biotechnological processes of keratin hydrolysis.

Functional Characterization of Genes Located at the Aurofusarin Biosynthesis Gene Cluster in Gibberella zeae

  • Kim, Jung-Eun;Kim, Jin-Cheol;Jin, Jian-Ming;Yun, Sung-Hwan;Lee, Yin-Won
    • The Plant Pathology Journal
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    • 제24권1호
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    • pp.8-16
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    • 2008
  • Aurofusarin is a polyketide pigment produced by some Fusarium species. The PKS12 and GIP1 genes, which encode a putative type I polyketide synthase (PKS) and a fungal laccase, respectively, are known to be required for aurofusarin biosynthesis in Gibberella zeae (anamorph: Fusarium graminearum). The ten additional genes, which are located within a 30 kb region of PKS12 and GIP1 and regulated by a putative transcription factor (GIP2), organize the aurofusarin biosynthetic cluster. To determine if they are essential for aurofusarin production in G. zeae, we have employed targeted gene deletion, complementation, and chemical analyses. GIP7, which encodes O-methyltransferase, is confirmed to be required for the conversion of norrubrofusarin to rubrofusarin, an intermediate of aurofusarin. GIP1-, GIP3-, and GIP8-deleted strains accumulated rubrofusarin, indicating those gene products are essential enzymes for the conversion of rubrofusarin to aurofusarin. Based on the phenotypic changes in the gene deletion strains examined, we propose a possible pathway for aurofusarin biosynthesis in G. zeae. Our results would provide important information for better understanding of naphthoquinone biosynthesis in other fdarnentous fungi as well as the aurofusarin biosynthesis in G. zeae.

Computational Approaches for Structural and Functional Genomics

  • Brenner, Steven-E.
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2000년도 International Symposium on Bioinformatics
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    • pp.17-20
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    • 2000
  • Structural genomics aims to provide a good experimental structure or computational model of every tractable protein in a complete genome. Underlying this goal is the immense value of protein structure, especially in permitting recognition of distant evolutionary relationships for proteins whose sequence analysis has failed to find any significant homolog. A considerable fraction of the genes in all sequenced genomes have no known function, and structure determination provides a direct means of revealing homology that may be used to infer their putative molecular function. The solved structures will be similarly useful for elucidating the biochemical or biophysical role of proteins that have been previously ascribed only phenotypic functions. More generally, knowledge of an increasingly complete repertoire of protein structures will aid structure prediction methods, improve understanding of protein structure, and ultimately lend insight into molecular interactions and pathways. We use computational methods to select families whose structures cannot be predicted and which are likely to be amenable to experimental characterization. Methods to be employed included modern sequence analysis and clustering algorithms. A critical component is consultation of the presage database for structural genomics, which records the community's experimental work underway and computational predictions. The protein families are ranked according to several criteria including taxonomic diversity and known functional information. Individual proteins, often homologs from hyperthermophiles, are selected from these families as targets for structure determination. The solved structures are examined for structural similarity to other proteins of known structure. Homologous proteins in sequence databases are computationally modeled, to provide a resource of protein structure models complementing the experimentally solved protein structures.

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Overexpression and characterization of thermostable chitinase from Bacillus atrophaeus SC081 in Escherichia coli

  • Cho, Eun-Kyung;Choi, In-Soon;Choi, Young-Ju
    • BMB Reports
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    • 제44권3호
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    • pp.193-198
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    • 2011
  • The chitinase-producing strain SC081 was isolated from Korean traditional soy sauce and identified as Bacillus atrophaeus based on a phylogenetic analysis of the 16S rDNA sequence and a phenotypic analysis. A gene encoding chitinase from B. atrophaeus SC081 was cloned in Escherichia coli and was named SCChi-1 (GQ360078). The SCChi-1 nucleotide sequences were composed of 1788 base pairs and 596 amino acids, which were 92.6, 89.6, 89.3, and 78.9% identical to those of Bacillus subtilis (ABG57262), Bacillus pumilus (ABI15082), Bacillus amyloliquefaciens (ABO15008), and Bacillus licheniformis (ACF40833), respectively. A recombinant SCChi-1 containing a hexahistidine tag at the amino-terminus was constructed, overexpressed, and purified in E. coli to characterize SCChi-1. $H_6SCChi$-1 revealed a hydrolytic band on zymograms containing 0.1% glycol chitin and showed the highest lytic activity on colloidal chitin and acidic chitosan. The optimal temperature and pH for chitinolytic activity were $50^{\circ}C$ and pH 8.0, respectively.

Positional Cloning and Phenotypic Characterization of a New Mutant Mouse with Neuronal Migration Abnormality

  • Park, Chankyu;Ackerman, Susan-L
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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    • pp.14-17
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    • 2001
  • Positional cloning (map-based cloning) of mutations or genetic variations has been served as an invaluable tool to understand in-vivo functions of genes and to identify molecular components underlying phenotypes of interest. Mice homozygous for the cerebellar deficient folia (cdf) mutation are ataxic, with cerebellar hypoplasia and abnormal lobulation of the cerebellum. In the cdf mutant cerebellum approximately 40% of Purkinje cells are ectopically located within the white matter and the inner granule cell layer (IGL). To identify the cdf gene, a high-resolution genetic map for the cdf-gene-encompassing region was constructed using 1997 F2 mice generated from C3H/HeSnJ-cdf/cdf and CAST/Ei intercross. The cdf gene showed complete linkage disequilibrium with three tightly linked markers D6Mit208, D6Mit359, and D6Mit225. A contig using YAC, BAC, and P1 clones was constructed for the cdf critical region to identify the gene. A deletion in the cdf critical region on chromosome 6 that removes approximately 150kb of DNA was identified. A gene associated with this deletion was identified using cDNA selection. cdf mutant mice with the transgenic copy of the identified gene restored the brain abnormalities of the mutant mice. The positional cloning of cdf gene provides a good example showing the identification of a gene could lead to finding a new component of important molecular pathways.

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