• Title/Summary/Keyword: Agar plate

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The Effect of Antioxidant and Antimicrobial Activity on the Extracted Its Material and Aloe vera L. Callus Culture by the Natural Fruit Juice (천연과즙을 이용한 Aloe vera L.의 callus 배양과 이들 추출물의 항산화 및 항균활성 효과 검정)

  • Lee, In-Soon;Bae, Dong Nyeok;Kwon, Oh Min;Han, Gu Tai;Kim, Dae Hwan;Oh, Myeong Won;Lee, Ji Hong;Moon, Hae-Yeon
    • KSBB Journal
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    • v.28 no.6
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    • pp.408-414
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    • 2013
  • In order to obtain functional materials from aloe callus, we cultured Aloe vera L. leaf on MS medium added 0.2 mg/L IAA, 0.3 mg/L kinetin and 100 mg/L grape or/and apple juice for 30 days. While a callus differentiation during callus culture did not show, the cultured leaves were uniquely released extracellular material into the agar plate. After cultivation for 18 days, the cultured leaf and agar were harvested for extraction a functional material. The materials extracted were measured on the amount of total phenols, flavonoids and polysaccharides and determined on the antioxidant and antimicrobial activity. In result, callus extracts of additive free (CT) and added apple juice (2T) had more amount of phenol compound ($659{\mu}g/mL$, $533{\mu}g/mL$) and flavonoid ($580{\mu}g/mL$, $501{\mu}g/mL$) than natural leaf (p: $525{\mu}g/mL$, f: $301{\mu}g/mL$). However, the extract of natural leaf had the better effect of lipid peroxidation and polysaccharide content than the culture extract. All samples extracted had same effect on the nitrite scavenging activity. On the other hand, only 2T extract showed excellent 72% nitric oxide scavenging activity. The agar extract was also confirmed to contain polyphenol compound and polysaccharide content that had antioxidant and antimicrobial activity partly.

Candidate of Probiotic Bacteria Isolated from Several Jeotgals: Korean Traditional Fermented Seafoods

  • Cho, Gyu-Sung;Do, Hyung-Ki;Bae, Chae-Yoon;Cho, Gyu-Sup;Whang, Cher-Won;Shin, Heuyn-Kil
    • Preventive Nutrition and Food Science
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    • v.11 no.2
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    • pp.140-145
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    • 2006
  • Seventy eight bacterial strains were isolated from several jeotgals using MRS and M 17 agar media. The probiotic properties such as tolerance of extreme growth condition, production of antimicrobial compound, production of hydrogen peroxide, and enzymatic activity of bile salt hydrolase were investigated. DHK 4, 10, 21 and 74 strains showed_a strong tolerance property against extreme conditions such as low pH and 0.5% oxgall-supplemented medium. DHK 10 and 47 strains produced hydrogen peroxide on TMB agar plate. DHK 8 and 10 strains produced antimicrobial compounds onto MRS agar against E. facalis. DHK 4, 6, 21, 29, 33, 63 and 87 strains had high activities of bile salt hydrolase. Especially, DHK 10 displayed a strong probiotic candidate; the abilities to produce the antimicrobial compound, hydrogen peroxide, and bile salt hydrolase. All these strains are assumed to be useful probiotic candidates. Among 78, twenty seven strains which have probiotic properties were tentatively identified by 16S rRNA sequencing. Among them, 7 Lactobacillus spp., 6 Leuconosotoc spp., 2 Weisella spp., 1 Pediococcus sp., 1 Staphylococcus sp., 1 Enterococcus sp. and 2 Streptococcus spp. were tentatively identified.

Screening of Cholesterol-lowering Bifidobacterium from Guizhou Xiang Pigs, and Evaluation of Its Tolerance to Oxygen, Acid, and Bile

  • Zhang, Rujiao;He, Laping;Zhang, Ling;Li, Cuiqin;Zhu, Qiujin
    • Food Science of Animal Resources
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    • v.36 no.1
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    • pp.37-43
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    • 2016
  • Cardiovascular and cerebrovascular diseases seriously harm human health, and Bifidobacterium is the most beneficial probiotic in the gastrointestinal tract of humans. This work aimed to screen cholesterol-lowering Bifidobacterium from Guizhou Xiang Pig and evaluate its tolerance to oxygen, acid, and bile. Twenty-seven aerotolerant strains with similar colony to Bifidobacterium were isolated through incubation at 37℃ in 20% (v/v) CO2-80% (v/v) atmospheric air by using Mupirocin lithium modified MRS agar medium, modified PTYG with added CaCO3, and modified PTYG supplemented with X-gal. Ten strains with cholesterol-lowering rates above 20% (w/w) were used for further screening. The selected strains’ tolerance to acid and bile was then determined. A combination of colony and cell morphology, physiological, and biochemical experiments, as well as 16S rRNA gene-sequence analysis, was performed. Results suggested that BZ25 with excellent characteristics of high cholesterol-removal rate of 36.32% (w/w), as well as tolerance to acid and bile, was identified as Bifidobacterium animalis subsp. lactis. To further evaluate Bifidobacterium BZ25’s growth characteristic and tolerance to oxygen, culture experiments were performed in liquid medium and an agar plate. Findings suggested that BZ25 grew well both in environmental 20% (v/v) CO2-80% (v/v) atmospheric air and in 100% atmospheric air because BZ25 reached an absorbance of 1.185 at 600 nm in 100% atmospheric air. Moreover, BZ25 was aerotolerant and can grow in an agar medium under the environmental condition of 100% atmospheric air. This study can lay a preliminary foundation for the potential industrial applications of BZ25.

Incompatibility Factors and Genetic Analysis of Pleurotus sajor-caju (여름느타리버섯의 교배계(交配系) 및 교배균주(交配菌株)의 유전(遺傳) 분석(分析))

  • Go, Seung-Joo;Shin, Gwan-Chull
    • The Korean Journal of Mycology
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    • v.14 no.1
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    • pp.17-23
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    • 1986
  • The mating system of monokaryotic isolates in Pleurotus sajor-caju was controlled by two incompatibility factors A and B of tetrapola mating system. The mycelia of dikaryotic isolates grew faster than those of their component monokaryons, but no correlation between dikaryotic and their component monokaryotic isolates was found. The primodia formed well on the potato dextrose agar (PDA) media not only in the dikaryotic isolates but also in the monokaryons under irradiated conditions. The dikaryotic isolates produced normal sporophores; however, the monokaryotic isolates produced abnormal sporophores when they were cultivated with sawdust substrates. Some dikaryotic isolates derived by mating between monokaryotic isolates showed high yields of sporophores more than those of parental strains. Both the dikaryotic mycelial growth rate and primodia formation number on the PDA plate showed significant correlation with its sporophore products on sawdust substrates.

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Isolation and Characterization of an Agarase-Producing Bacterial Strain, Alteromonas sp. GNUM-1, from the West Sea, Korea

  • Kim, Jonghee;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1621-1628
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    • 2012
  • The agar-degrading bacterium GNUM-1 was isolated from the brown algal species Sargassum serratifolium, which was obtained from the West Sea of Korea, by using the selective artificial seawater agar plate. The cells were Gram-negative, $0.5-0.6{\mu}m$ wide and $2.0-2.5{\mu}m$ long curved rods with a single polar flagellum, forming nonpigmented, circular, smooth colonies. Cells grew at $20^{\circ}C-37^{\circ}C$, between pH 5.0 and 9.0, and at 1-10% (w/v) NaCl. The DNA G+C content of the GNUM-1 strain was 45.5 mol%. The 16S rRNA sequence of the GNUM-1 was very similar to those of Alteromonas stellipolaris LMG 21861 (99.86% sequence homology) and Alteromonas addita $R10SW13^T$(99.64% sequence homology), which led us to assign it to the genus Alteromonas. It showed positive activities for agarase, amylase, gelatinase, alkaline phosphatase, esterase (C8), lipase (C14), leucine arylamidase, valine arylamidase, ${\alpha}$-chymotrypsin, acid phosphatase, naphthol-AS-BI-phosphohydrolase, ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\beta}$-glucosidase, catalase, and urease. It can utilize citrate, malic acid, and trisodium citrate. The major fatty acids were summed feature 3 (21.5%, comprising $C_{16:1}{\omega}7c/iso-C_{15:0}$ 2-OH) and C16:0 (15.04%). On the basis of the variations in many biochemical characteristics, GNUM-1 was considered as unique and thus was named Alteromonas sp. GNUM-1. It produced the highest agarase activity in modified ASW medium containing 0.4% sucrose, but lower activity in rich media despite superior growth, implying that agarase production is tightly regulated and repressed in a rich nutrient condition. The 30 kDa protein with agarase activity was identified by zymography, and this report serves as the very first account of such a protein in the genus Alteromonas.

A study on improvement of 2. 3. 5-Triphenyl Tetrazolium Chloride(TTC) reduction test : disc plate method (2. 3. 5-Triphenyl Terazolium Chloride(TTC) 환원시험법의 개선에 관한 연구 : disc plate method)

  • 정동수;김동훈;이상목
    • Korean Journal of Veterinary Service
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    • v.18 no.2
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    • pp.163-176
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    • 1995
  • Triphenyl Tetrazolium Chloride(TCC) reduction test is simple and sensitive to some residual antibiotics (especially to penicillin) in milk, but comparatively insensible to sulfo-namides. The volumn of sample is also large. Thus this study was undertaken to increase the detectable level of sulfonamides in raw milk. In this study, we used small transparent plastic hole and pulp disc instead of 10m1 test tube and made test medium in which was added 0.08%TTC, 0.3% agar, 10% skim milk, approximately $10^6$ CFU/ml streptococcus thermophilus and 5ppm Trimethoprim to enhance the sensitivity for sulfonamides The results of TCC reduction test by disc plate method were summarized as follows : 1. sensitivity to residual sulfonamides were much higher than official TCC reduction test. Detectable limites of sulfamethazine, sulfamerazine, sulfathiazole, sulfachloropy-ridazine, sulfadimethoxine, sulfamononethoxine, sulfadiazine and sulfaquinoxaline were 0.1-0.5ppm levels. 2. Detectable limites to some antibiotics were simillar or good than that of official method as 0.005-0.1ppm to three ${\beta}$ -lactams, 0.25-0.5ppm to one macrolide, 2-10ppm to three aminoglycosides, 0.2-0.5ppm to three tetracycline, 0.1-0.5ppm to chloramphenicol. 3. Only 0.1ml of milk was needed to test and the test medium could be stored appnoximatly 7days in the refrigerator. So test procedure was convenient than offcial method. 4. These results suggest that disc plate method is more useful to detect bacterial growth inhibition substances including sulfonamides in raw milk.

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Cloning and High Expression of Nattokinase Gene from Bacillus subtilis BB-1 (Bacillus subtilis BB-1으로부터 나토키나아제 유전자 크로닝 및 대량발현)

  • Lee Young-Hoon;Lee Sung-Ho;Park Ki-Hoon;Choi Young-Ju;Jeong Yong-Kee;Gal Sang-Wan
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.274-281
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    • 2006
  • A fibrinolytic enzyme gene was isolated from Bacillus subtilis BB-1 by PCR method. Primers for PCR cloning were designed according to pre-identified gene for fibrinolytic enzymes from B. subtilis. The primer sequences were 5'-CGG ATC CGT GAG AGG CAA AAA GGT G-3' and 5'-TGA ATT CTT AAT GTG CTG CTG CTT GTC C-3' as concensus sequences of the fibrinolytic genes of Bacillus species. The PCR product was 1,145 bp and the sequence homology was 99% with nattokinase gene isolated from Japanese natto. The cloned fibrinolytic gene was reconstructed in Bacillus-E. coli shuttle vector, pEB for bulk-production. The fibrinolytic enzyme was purified by FPLC from the cloned B. subtilis 168. The optimum pH and temperature of the enzyme were 7.0 and $35^{\circ}C$, respectively. The fibrinolytic enzyme did not show any activity toward to skim milk, gelatin, casein and blood agar plate. The enzyme specific polyclonal antibody was prepared in rabbit for further assays such as detection of the gene expression in plant cells. This means that the enzyme may be used for health-care such as thrombosis without any hamful effects in the blood vessel.

Isolation and Characterization of Microorganisms with Broad Antifungal Activity against Phytopathogenic Fungi (식물병원균에 광범위 항균활성을 가진 미생물의 분리 및 특성)

  • Kim, Min-Hee;Ko, Hee-Sun;Yook, Young-Min;Kim, Hyun-Soo
    • KSBB Journal
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    • v.23 no.3
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    • pp.219-225
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    • 2008
  • For the production of an antifungal compound, one strain (I-8) was selected from approximately 400 strains isolated from various soil samples. The optimum carbon source, nitrogen source and pH culture conditions for the production of the antifungal compound were investigated. ISP No. 2 medium (yeast extract 0.4%, malt extract 1% and dextrose 0.4%, at pH 8) was determined to be the optimum medium. Strain I-8 showed broad antifungal activity against the plant pathogenic fungi tested, including Sclerotinia sclerotiorum KACC 41065, as well as cellulase and chitinase activities in an agar plate assay. The extraction of antifungal compounds was performed using ethyl ether and ethyl acetate. In a culture broth of strain I-8, the ethyl acetate extract exhibited effective growth inhibition against 14 of the 20 phytopathogenic fungi tested. By mixing the ethyl acetate extract from I-8 with the ethyl ether extract from the fungus 13-16, which shows specific antifungal activity against Colletotrichum orbiculare KACC 40808, the antifungal activity of I-8 against phytopathogenic fungi was confirmed to be slightly increased. Strain I-8 showed strong growth inhibition against 16 phytopathogenic strains in agar plate tests.

Effect of Growth Medium and Extract Sterilization on a Seedling Bioassay of Alfalfa Autotoxicity (생장배지 및 추출액의 멸균이 알팔파 Autotoxicity 의 유묘 생물검정에 미치는 영향)

  • Chon, Sang-Uk;Nelson, C.Jerry;Han, Sung-Uk;Guh, Ja-Ock
    • Korean Journal of Environmental Agriculture
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    • v.18 no.2
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    • pp.185-190
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    • 1999
  • Autotoxic substance(s) from alfalfa(Medicago sativa L.) plants reduces germination and growth of adjacent new alfalfa after alfalfa. The autotoxic chemical(s) in alfalfa are clearly unknown. Our objective was to improve the sensitivity of an alfalfa seedling bioassay for evaluating autotoxic leaf extracts. We determined critical extract concentrations that inhibit seed germination and seedling growth, compared two different culture media, and evaluated the effects of extract sterilization on the sensitivity of the assay, by using streptomycin and autoclaving method. An agar medium in petri plate gave better responses of germination and seedling growth to the extracts than using filter paper in the plate. On agar medium, the concentration of extract required to reach 50% inhibition of root length was 2.7 g $kg^{-1}$, and of germination and hypocotyl length were 3.8 and 9.9 g $kg^{-1}$, respectively. Leaf extracts with 100 ppm streptomycin stimulated germination significantly compared to Leaf extract alone but reduced root length of control by 43%. Root length was more sensitive to the autotoxin(s) than was germination or hypocotyl length. These results suggest that agar medium mixed with extract and sterilization by autoclaving could be improved the consistency and precision of bioassay, and that root length was the best parameter of autotoxic effect of alfalfa leaf extract.

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IN VITRO GROWTH OF CANDIDA ALBICANS ON SEVERAL RESILIENT DINTURE LINERS (수종의 탄성 의치상 이장재에 대한 Candida albicans의 성장에 관한 연구)

  • Chung, Chae-Heon;Kim, Kwang-Won;Kim, Dong-Ki;Lee, Zang-Hee
    • The Journal of Korean Academy of Prosthodontics
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    • v.31 no.1
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    • pp.19-27
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    • 1993
  • For the purpose of this study was to determine the growth of Candide albicans on the surface of the resilient denture liners. The discs$(40\times40mm)$ of 2 resilient lining materals (Molloplast B, Mollosil) and one conventional acrylic resin (K-33) and one metal plate were processed and disinfected. Firstly, the test discs were placed into petri dish, and Candide albicans suspensions was overlayed on the test discs. And the test discs were incubated with intermitant shaking for 1 hour, 2 hours, 6 hours, 12 hours, 24 hours. After incubation, imprint culture method was achived and counted the colony on the agar plate. Secondly, the effect of denture cleansing agents on the growth of Candide alibicans on the resilient dentureliners was evaluated. The results were as follows : 1. The growth of Candida albicans on discs of Molloplast B and Mollosil was increased than that on discs of acrylic resin and metal plate (p<0.05). 2. As Candide albicans suspensions were incubated for 2 hours, the growth of Candida albicans on discs of Mollosil was increased than that on discs of Molloplast B (p<0.05), and the growth of Candide albicans on discs of metal plate was increased than that on discs of acrylic resin (p<0.05). 3. As Candide albicans suspensions were incubated for 6 hours, the growth of Candide albicans on discs of Mollosil was increased than that on discs of Molloplast B (p<0.05). 4. The growth of Candide albicans on discs of Mollosil and Molloplast B in treating denture cleansing agent was inhibited than control discs (p<0.05).

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