• Title/Summary/Keyword: High isolation

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Isolation and Partial Characterization of Isoflavone Transforming Lactobacillus plantarum YS712 for Potential Probiotic Use (Isoflavone 비배당화 및 항산화 활성을 지닌 Lactobacillus plantarum YS712의 선발)

  • Cho, Yoon-Hee;Imm, Jee-Young;Kim, Hwa-Young;Hong, Seong-Gil;Hwang, Sung-Joo;Park, Dong-Jun;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.29 no.5
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    • pp.640-646
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    • 2009
  • Lactic acid bacteria (LAB) are typical probiotic microbes that are used in various industries including fermented foods, feed additives, and pharmaceuticals. The purpose of this study was to compare the ability of isoflavone biotransformation and antioxidative activity of 17 LAB. Six strains including the Lactobacillus species exhibited a 100% hydrolysis rate for daidzein during fermentation. The content of total genistein in soymilk fermented with these strains was $872-943\;{\mu}g/g$. The DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging ability of the LAB was widely variable and ranged from 23-78%. A selected strain was isolated from kimchi and the strain was identified as Lactobacillus plantarum ssp. through the API carbohydrate fermentation pattern and 16S rDNA profile. The strain exhibited excellent acid tolerance in an artificial gastric solution. L. plantarum YS712 showed high $\beta$-glucosidase activity in fermentation. The concentration of genistein and daidzein in soymilk fermented with L. plantarum YS712 increased from 3.64 to $917.3\;{\mu}g/g$ and from 58.18 to $1062.17\;{\mu}g/g$, respectively. These results demonstrate the potential of L. plantarum YS712 as a probiotic culture that can be utilized in the manufacturing of fermentation foods and dietary supplements.

Isolation, Characterization, and Molecular Cloning of the cDNA Encoding a Novel Phytase from Aspergillus niger 113 and High Expression in Pichia pastoris

  • Xiong, Ai Sheng;Yao, Quan-Hong;Peng, Ri-He;Li, Xian;Fan, Hui-Qin;Guo, Mei-Jin;Zhang, Si-Liang
    • BMB Reports
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    • v.37 no.3
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    • pp.282-291
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    • 2004
  • Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.

Isolation and Identification of Antagonistic Bacterium Active against Sclerotinia sclerotioum Causing Sclerotinia Rot on Crisphead Lettuce (결구상추 균핵병균(Sclerotinia sclerotioum)에 대한 길항세균의 분리 및 동정)

  • Kim, Han-Woo;Lee, Kwang-Youll;Baek, Jung-Woo;Kim, Hyun-Ju;Park, Jong-Young;Lee, Jin-Woo;Jung, Soon-Je;Moon, Byung-Ju
    • Research in Plant Disease
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    • v.10 no.4
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    • pp.331-336
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    • 2004
  • The fungus genus Sclerotinia contains a number of important plant pathogens. Vegetable growers in our country are probably most familiar with Sclerotinia sclerotiorum, the causes of sclerotinia rot on crisphead lettuce. S. sclerotiorum has a wide host range which can include lettuce as well as crops such as broccoli, cabbage, carrots, celery, beans, peppers, potatoes, stocks, and tomato. Some fungicides, including benomyl, are effective in some crops, but not all. So, we isolated a antagonistic bacteria that are active on sclerotinia rot caused by S. sclerotiorum and that can be used to control it. About 702 strains had been isolated from soil around plant roots in the field. Ten strains showed strong antifungal activity against S. sclerotiorum. In pot test for antagonistic activity, A-7 strain showed high control value against the pathogen when compared with others. The strain was, therefore, selected as a biocontrol candidate against sclerotinia rot and its biochemical properties and 16S rDNA sequence was analyzed. The A-7 strain was highly related to Bacillus subtilis and B. amyloliquefaciens. To confirm precise identification, we had performed gyr A gene sequences analysis. Its sequence had 96% similarity with B. amyloliquefaciens. Consequently, the isolate was identified as B. amyloliquefaciens A-7.

Epizootiological Investigation on Infectious Laryngotracheitis in Commercial Chicken Flocks (닭 전염성후두기관염의 역학적 조사)

  • 김선중;서익수
    • Korean Journal of Poultry Science
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    • v.10 no.2
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    • pp.113-121
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    • 1983
  • In an effort to understand epizootiological aspects of infectious laryngotracheitis (ILT), a total of 56 chicken flocks in six farms comprised of 35 broiler breeder, 13 commercial layer and 6 layer breeder flocks. were investigated. The farms experienced ILT during the period of one year from June, 1982. In most farms the birds were vaccinated against ILT just before or after the disease outbreak. In two of the farms in which ILT broke out in winter, it was possible to contain the disease in only one or two fleets without transmitting it to the remaining 5 to 7 flocks in the farms by adopting strict isolation procedures for the affected flocks. In regarding inter- flock spreading speed, it took an average of 6 days for flocks rearing on floor and 11 days for those in cages. Among the flecks in rearing cages. transmission among laying flocks was much faster. taking an average of 8 days, compared to non-laying flocks of 17 days, suggesting spreading of the disease by means of egg trays or egg collection process. Peak mortality was observed between 5 and 10 days after from the time of appearance of first dead birds from the disease and the period of mortality, with an average of 18 days, was not influenced by rearing systems, breeds and age of birds. Mortalities in the affected flocks ranged from lo/e to 19.8%, with an average of 6.5 %, and was also not influenced by the above variables except significantly lower mortality in immature broiler breeder flocks (2.9%) compared to immature layer (11.8%) and mature broiler breeder flocks (6.9%). In one breeder farm in which all the birds were kept on floor and ILT broke out in summer, mortality in male birds in all seven flocks of 37 weeks of age or older was as high as twice of that in female birds in the same flocks. This trend was not observed in one 31 weeks old flock and was reversed in another 14 weeks old flock in the farm.

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Isolation and Characterization of ACC Synthase Gene Family in Mung Bean (Vigna radiata L.): Differential Expression of the Three ACC Synthase enes in Response to Auxin and Brassinosteroid

  • Sunjoo Joo;Kim, Woo-Taek
    • Journal of Plant Biotechnology
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    • v.2 no.2
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    • pp.61-71
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    • 2000
  • By screening a cDNA library of auxin-treated mung bean (Vigna radiata L.) hypocotyls, we have isolated two full-length cDNA clones, pVR-ACS6 and pVR-ACS7, for 1-aminocyclopropane-1-carboxylate (ACC) synthase, the rate-limiting enzyme in the ethylene biosynthetic pathway. While PVR-ACS6 corresponds to the previously identified PCR fragment pMBA1, pVR-ACS7 is a new cDNA clone. A comparison of deduced amino acid sequences among auxin-induced ACC synthases reveal that these enzymes share a high degree of homology (65-75%) to VR-ACS6 and VR-ACS7 polypeptides, but only about 50% to VR-ACS1 polypeptide. ACS6 and ACS7 are specifically induced by auxin, while ACS1 is induced by cycloheximide, and to lesser extent by excision and auxin treatment. Results from nuclear run-on transcription assay and RNA gel blot studies revealed that all three genes were transcriptionally active displaying unique patterns of induction by IAA and various hormones in etiolated hypocotyls. Particularly, 24-epibrassinolide (BR), an active brassinosteroid, specifically enhanced the expression of VR-ACS7 by distinct temporal induction mechanism compared to that of IAA. In addition, BR synergistically increased the IAA-induced VR-ACS6 and VR-ACS7 transcript levels, while it effectively abolished both the IAA- and kinetin-induced accumulation of VR-ACS1 mRNA. In light-grown plants, VR-ACS1 was induced by IAA in roots, whereas W-ACS6 in epicotyls. IAA- and BR-treatments were not able to increase the VR-ACS7 transcript in the light-grown tissues. These results indicate that the expression of ACC synthase multigene family is regulated by complex hormonal and developmental networks in a gene- and tissue-specific manner in mung bean plants. The VR-ACS7 gene was isolated, and chimeric fusion between the 2.4 kb 5'-upstream region and the $\beta$-glucuronidase (GUS) reporter gene was constructed and introduced into Nicotiana tobacum. Analysis of transgenic tobacco plants revealed the VR-ACS7 promoter-driven GUS activity at a highly localized region of the hypocotyl-root junction of control seedlings, while a marked induction of GUS activity was detected only in the hypocotyl region of the IAA-treated transgenic seedlings where rapid cell elongation occurs. Although there was a modest synergistic effect of BR on the IAA-induced GUS activity, BR alone failed to increase the GUS activity, suggesting that induction of VR-ACS7 occurs via separate signaling pathways in response to IAA and BR.

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Epidemiological Observation on Recent Outbreaks of Canine Parvoviral Enteritis in Korea (최근 국내발병 개 파보바이러스성 장염에 대한 역학적 조사)

  • Jeoung, Seok-Young;Kim, Doo;Ahn, So-Jeo;Pak, Son-Il
    • Journal of Veterinary Clinics
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    • v.23 no.3
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    • pp.223-229
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    • 2006
  • Recently canine parvovirus(CPV) enteritis had been controlled successfully by the use of low-passage high titer modified live CPV vaccines. However, outbreaks of CPV enteritis have been continued in Korea. In this study, we carried out epidemiological investigation on the recent outbreaks of CPV enteritis of dogs and determined the potential prognostic factors affecting the survival of dogs. The total of 140 dogs diagnosed for CPV enteritis were statistically analysed. The majority of dogs were from 6 to 18 weeks of age and were not vaccinated or incompletely vaccinated. There were significant difference in the survival rate between male and female dogs with CPV enteritis and among the age groups(p<0.05). Moreover, there was significant difference in the survival rate of dogs between HI titer <80 group and HI titer 160 group(p<0.05). The majority of dogs had a history of diarrhea, vomiting, lethargy and dehydration. It was considered that recent outbreak of CPV enteritis in Korea caused by the failure of vaccination and/or by the inadequate antibody responses to CPV vaccines. Prophylactic measures should include isolation of young dogs from the dog population until the vaccination can be expected to provide protection.

Seasonal Variation of Redox Potential in Jinkwannaedong Ecological Conservation Area (진관내동 생태계보전지역에서 산화환원전위(Redox Potentia)의 월별 변화)

  • Kim, Jae-Geun
    • Journal of Wetlands Research
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    • v.6 no.2
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    • pp.65-71
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    • 2004
  • The most significant effect of excess water in wetlands is the isolation of the soil from the atmosphere and the prevention of O2 from diffusing into soil. The blockage of atmospheric O2 induces biological and chemical processes that change soil from oxidized into reduced state. When dry soil develop into hydric soil, redox potential is dropping. The redox potential is a indicator of hydric soil and affect chemical function of wetlands. To reveal characteristics of wetland soil, redox potential was measured in Jinkwannaedong ecological conservation area from May in 2003 to March in 2004. Redox potentials in May ranged from 5 mV at 25 cm depth to 200 mV at 10 cm depth. It decreased to about -200 m V at all depths and continued until October. In winter, redox potential was slowly increased; it was the highest at 5 cm depth and lowest at 20 cm depth. Annual variations of redox potential in 20 cm depth showed the same pattern at 5 sites; low in growing season and high in non-growing season. This results indicates that soils of study sites are in hydric state and methanogenesis is occurring in Jinkwannaedong ecological conservation area.

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Isolation and Characterization of Bacteria Capable of Degrading Bisphenol A (Bisphenol A 분해세균의 분리 및 특성)

  • 김희식;이영기;이완석;박찬선;윤병대;오희목
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.189-196
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    • 2001
  • Eighty-seven microbial strains capable of growing on bisphenol A (BPA) as a sole carbon source were isolated from soils, waste waters and sludges. Among them, three bacterial strains were finally selected as potential decomposers through measuring BPA-degradation efficiency by HPLC analysis. Two of these bacterial strains were identified as Serratia marcescens 1901 and S. marcescens 1902, and another was Pseudomonas putida 1401 by 16S rDNA partial sequences and based on morphological and physiological properties. They showed higher cell growth and BPA degradation in PAV (PAS medium containing vitamin mixtures) than in PAS medium. The degradation efficiencies of these bacterial strains were within a range of 20-40% in the PAV containing 500 mg/1 or 100 mg/l of BPA fur 3 days. S. marcescens 1901 showed higher degradation efficiency at 100 mg/1 of BPA than those of other selected strains, while S. marcescens 1902 and P. putida 1401 degraded a high concentration of BPA (500 mg/l) with a degradation efficiency of 40% for 3 days. The BPA degradation using a mixed culture of three selected strains showed the similar level of dog-radation efficiency with that using a pure culture.

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Seasonal Changed of Microbial Population in the Field Soil (계절에 따른 토양중 미생물의 밀도 변화)

  • Park, Dong-Jin;Lee, Sang-Hwa;Kim, Chang-Jin
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.144-148
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    • 1998
  • Soil microorganisms including bacteria, fungi, and actinomycetes were seasonally isolated at depths (0.5~2, $10{\pm}1$, $50{\pm}1cm$) of field. The frequency of microbial isolates was employed for the determination of microbial population (CFU/g dry soil) and distribution ratio (%) in soil. Both bacteria (24-fold) and actinomycetes (7-fold) exhibited the biggest change at the depth of $50{\pm}1cm$, whereas fungi showed the maximum (13-fold) at $10{\pm}1cm$. On the whole, the bacterial population was high in spring soil, fungi in winter, and actinomycetes in autumn. Soil microorganisms also exhibited the seasonal variation on their distribution ratio (%). The maximum distribution ratio (85.7%) of bacteria was observed at the depth of $50{\pm}1cm$ in spring, whereas bacteria showed the minimum (35.2%) at the depth of $10{\pm}1cm$ in spring. The maximum distribution ratio (23.0%) of fungi was found at the depth of $50{\pm}1cm$ in spring, whereas its minimum (0.5%) at the depth of $10{\pm}1cm$ in spring. Actinomycetes exhibited the maximum distribution ratio (45.2%) at the depth of $10{\pm}1cm$ in spring, whereas its minimum (12.2%) was showed at the depth of $50{\pm}1cm$ in spring.

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Isolation and characterization of a novel DNA segment that enables the plasmids to replicate autonomously in Aspergillus nidulans (Aspergillus nidulans에서 플라스미드의 자가복제를 유발하는 DNA절편의 분리 및 분석)

  • Kim, Jin-Hee;Han, Kyu-Yong;Han, Kap-Hoon;Han, Dong-Min
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.120-125
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    • 1998
  • A plasmid pNPG contains a genomic DNA complementing npgA1 which is located on the left arm of linkage group I. It transformed Aspergillus nidulans at a high frequency. No abortive transformants were observed and the $Trp^+$ transformants were all $Npg^+$. The 10.4 kb Psti fragment of the genomic DNA was subcloned into pILJ16, which increased the transformation efficiency by more than 200-folds. The transformants were mitotically unstable and yielded $Arg^-$ conidia at the frequency of more than 80%. An additional gene cloned into the plasmid containing the fragment was always lost with $argB^+$ marker. These characteristics strongly indicate the possibility that the plasmids autonomously replicate. The full activity of enhanced transformation was retained on the 4.9 kb EcoRI-HaeIII fragment. The DNA segment was similar to AMA1 rather than ANS1 in function and designated AMA2.

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