• Title/Summary/Keyword: Agar Gel

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Overexpression of Arylsulfatase in E. coli and Its Application to Desulfatation of Agar

  • Lim, Jae-Myung;Jang, Yeon-Hwa;Kim, Hyeung-Rak;Kim, Young-Tae;Choi, Tae-Jin;Kim, Joong-Kyun;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.777-782
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    • 2004
  • The arylsulfatase gene (astA, 984 bp ORF) from the P. carrageenovora genome was amplified by PCR and subcloned into the pET21a vector. When the constructed plasmid pAST-A1 (6.4 kb) was introduced into E. coli BL21(DE3), the transformant on the LB plate containing IPTG showed a hydrolyzing activity for 4-methylumbelliferyl sulfate and p-nitrophenyl sulfate. The highest arylsulfatase activity (2.1 unit/ml) was obtained at 10 mM IPTG. Most arylsulfatase activity was found in the cell lysate, whereas no significant activity was detected in the culture supernatant. The molecular weight of the recombinant enzyme was estimated to be 33.1 kDa by SDS-PAGE. After the reaction of agar with arylsulfatase for 12 h at $40^{\circ}C$, the gel strength of the agar increased by 2-fold, and 73% of the sulfate in the agar had been removed. This result suggests that arylsulfatase expressed in E. coli could be useful in the production of electrophoretic grade agarose.

Purification and Properties of a Novel Extracellular Agarase from Marine Bacterium, Sphingomonas paucimobilis AS-1 (해양미생물 Sphingomonas paucimobilis AS-1이 생산하는 새로운 extracelluar agarase의 정제 및 특성)

  • Jung, Il-Sun;Kim, Yu-Jung;Song, Hyo-Ju;Gal, Sang-Wan;Choi, Young-Ju
    • Journal of Life Science
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    • v.18 no.1
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    • pp.103-108
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    • 2008
  • An agar-degrading marine bacterium, strain AS-1 was isolated from the seawater. The strain AS-1 was identified as Sphingomonas paucimobilis (90% probability) by VITEK. The optimum medium for agarase activity of the isolated strain was determined to be marine medium, marine broth 2216 containing 0.1% agar as carbon source. An extracellular agarase was purified 104-fold from the culture supernatant by ammonium sulfate precipitation, ion exchange chromatography and gel filtration methods. The molecular weight of the purified enzyme was estimated to be 80 kDa by SDS-PAGE. The optimum pH and temperature for activity were 7.0 and $40^{\circ}C$, respectively. Antioxidative activity of the strain AS- was 72% in the supernatant cultured for 12 h. The culture supernatant of the strain AS-1 showed antibacterial activity against bacteria causing putrefaction and food poisoning such as Escherichia coli, Staphylococcus aureus and Proteus vulgaris. However, the cell growth of the lactic aicd forming strain, Lactobacillus plantarium was promoted by the treatment of 10% culture supernatant of an agar-degrading strain.

Characterization and Purification of Agarase from Cytophaga sp. ACLJ-18 (한천 분해균(Cytohaga sp. ACLJ-18)이 생산하는 agarase의 정제 및 특성)

  • 주동식;송해미;이정석;조순영;이응호
    • KSBB Journal
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    • v.13 no.3
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    • pp.320-324
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    • 1998
  • Agar degrading enzyme-agarase-was purified from the culture fluid of Cytophaga so/ ACLJ-18, by acetone precipitation, DEAE-Cellulose, Sephadex G-100 and CM-Sephadex C25 column chromatographies. The molecular weight of purified agarase was estimated to be 24,700 dalton by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for agarase activity were 7.0 and 40$^{\circ}C$, respectively. this agarase was stable in the pH range of 6.5 - 8.0 and 40$^{\circ}C$, and required 0.35M NaCl for optimum activity. And this agarase was inhibited by metal ions such as Ba2+, Cu2+, Co2+, Mn2+, Hg2+, Zn2+, and showed specificity on agar.

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Isolation and Characterization of a Novel Agarase-Producing Pseudoalteromonas spp. Bacterium from the Guts of Spiny Turban Shells

  • Oh, Young-Hoon;Jung, Chang-Kyou;Lee, Jin-Won
    • Journal of Microbiology and Biotechnology
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    • v.21 no.8
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    • pp.818-821
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    • 2011
  • An agar-degrading bacterium was isolated from the guts of spiny turban shells. It was identified as a Pseudoalteromonas species and named Pseudoalteromonas sp. JYBCL 1. The viscosity of the inoculated agar medium decreased by more than 60% after 20 h cultivation. The agarase produced by the isolate had optimal activities at $35^{\circ}C$ and pH 7. The enzyme had extremely strong resistance to ionic stress compared with other known agarases. Its molecular mass was estimated at about 60 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The agarase could saccharify Gelidium amansii directly, with an efficiency about half that compared with agar saccharification.

Analysis of Vaginal Lactic Acid Producing Bacteria in Healthy Women

  • Nam, Hye-Ran;Whang, Kyung-Hee;Lee, Yeon-Hee
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.515-520
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    • 2007
  • Vaginal lactic acid-producing bacteria of 80 pre-menopausal women were studied by isolation on Blood and DeMan-Rogosa-Sharpe agar, PCR with group-specific primers for Lactobacillus-denaturing gradient gel electrophoresis (DGGE), and PCR with specific primers for V3 region in 16S rRNA-temporal temperature gel electrophoresis (TTGE). Conventional isolation method on media detected only one lactobacillus (Lactobacillus brevis) while TTGE detected only Lactobacillus sp. DGGE detected seven Lactobacillus species; L. coleohominis, L. crispatus, L. iners, L. reuteri, L. rhamnosus, L. vaginalis, and Leuconostoc lactis. L. acidophilus and L. gasseri, which are prevalent in Western women, were not detected in Korean women. Furthermore, L. rhamnosus, Leuc. lactis, L. coleohominis, and Weissella cibaria, which were not previously reported in the vaginal microbiota of Korean women, were detected. The five most prevalent LABs in vaginal microbiota in Korean women were L. iners, Enterococcus faecalis, L. crispatus, Leuc. lactis, and W. cibaria.

Development of Formaldehyde-shielding Chitosan-gel Sheet (키토산 겔을 이용한 포름알데하이드 차폐 시트 개발)

  • Kim, So-Yeon;Kim, Mi-So;Tak, Sang Min;Lee, Ji Whan;Sim, So Yeon;Joo, Eun Hee;Kim, Sung Bae;Kim, Chang-Joon
    • KSBB Journal
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    • v.29 no.3
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    • pp.183-187
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    • 2014
  • Sick-building syndrome occurs when indoor air is polluted with harmful volatile organic compounds such as formaldehyde which are contained in furniture or new building materials. In this study, formaldehyde-shielding chitosangel sheet was developed and its performance was evaluated. Chitosan and agar were dissolved in acetic acid solution. The optimal concentrations of chitosan, acetic acid and agar were 3, 3, and 2.5 %(w/w). Formaldehyde was spreaded on gypsum board and then wall paper was attached on it by using glue. When chitosan-gel sheet was attached on this control board, the amount of formaldehyde released from the board was decreased by 63% than in control board. On the other hand, decrease in formaldehyde releasing was only 32% when liquid solution of chitosan was spreaded on the control board. This result clearly indicates that chitosan-gel sheet removes formaldehyde more effectively than liqud solution of chitosan. Furthermore, this type of sheet is more applicable to new building than spraying type.

Dissimilarity between Human and Bacterial DNA (사람의 DNA와 박테리아의 DNA 사이의 非類似性)

  • In Won Park
    • The Korean Journal of Zoology
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    • v.11 no.3
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    • pp.83-84
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    • 1968
  • An attempt was made to estimate the possible homology between human and bacterial DNA by the method of DNA-agar gel hybridization. HeLa DNA was embedded in the agar and $^14C-DNA$ Xanthomonas pelargonii was used as bacterial DNA for the sheared fragments. No homology between human and bacterial DNA was detected. If homology exists at all, it can be estimated from the sensitivity of the method and assuming some 1,000 nucleotide pairs per cistron, the not more than $2\times10^5$ base pairs or 200 bacterial cistrons would be preserved in human DNA, corresponding to less than 0.01 per cent of the total human genome.

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Investigations on the Virus Diseases in Spinach (Spinacia oleracea L.) II. Identification of Broad Bean Wilt Virus Occuring Spinach (시금치 바이러스병에 관한 연구 II. 시금치에 발생하는 Broad Bean Wilt Virus (BBWV)의 분류동정)

  • Lee S. H.;Lee K. W.;Chung B. J.
    • Korean journal of applied entomology
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    • v.18 no.1 s.38
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    • pp.11-14
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    • 1979
  • Spinaches showing dark green mosaic symptoms were used for identification of broad bean wilt virus. In host reaction test, that virus caused local lesions on the inoculated leaves and mosaic symptoms on upper leaves of Chenopodium amaranticolor, Chenopodium quinoa and Vicia faba, and developed mosaic symptoms on Physalis floridana, Spinacia oleracea, Nicotiana tabacum, (White burley, Bright yellow) Nicotiana glutinusa. In agar gel-diffusion test, the virus showed positive reaction with broad bean wilt virus antiserum. Spherical virus particles with size of 25nm in diameter were observed in electron microscope.

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Hydrocolloids Decrease the Digestibility of Corn Starch, Soy Protein, and Skim Milk and the Antioxidant Capacity of Grape Juice

  • Yi, Yue;Jeon, Hyeong-Ju;Yoon, Sun;Lee, Seung-Min
    • Preventive Nutrition and Food Science
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    • v.20 no.4
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    • pp.276-283
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    • 2015
  • Hydrocolloids have many applications in foods including their use in dysphagia diets. We aimed to evaluate whether hydrocolloids in foods affect the digestibility of starch and protein, and their effects on antioxidant capacity. The thickening hydrocolloids: locust bean gum and carboxymethyl cellulose, and the gel-forming agents: agar agar, konjacglucomannan, and Hot & Soft Plus were blended with corn starch and soy protein, skim milk, or grape juice and were examined for their in vitro-digestability by comparing the reducing sugar and trichloroacetic acid (TCA)-soluble peptide, for antioxidant capacity by total polyphenol contents and the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity. The hydrocolloids resulted in a decrease in starch digestibility with the gel-forming agents. Hydrocolloids diminished TCA-soluble peptides in skim milk compared to soy protein with the exception of locust bean gum and decreased free radical scavenging capacities and total phenolic contents in grape juice. Our findings may provide evidence for the use of hydrocolloids for people at risk of nutritional deficiencies such as dysphagia patients.

Transport and Distribution of Calcium Salt in Tofu Manufacturing Process -Part I. Conductometric Measurement of Calcium Salt in Tofu and Drained Solution- (두부제조공정(製造工程)중 Calcium 염(鹽)의 행동(行動)과 분포(分布) -제1보(第一報). 전기전도도법(電氣傳導度法)에 의한 두부 및 순물의 Calcium 농도 측정방법(測定方法)-)

  • Yim, Sang-Bin;Lee, Choon-Ki;Chun, Jae-Kum
    • Applied Biological Chemistry
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    • v.28 no.1
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    • pp.8-12
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    • 1985
  • For measuring calcium content during the manufacturing process of Tofu, conductometric electrodes were made with the copper plate, and the results were compared with chemical analysis methods. Three types of plate material (I, II, III) for electrode were tested to apply for measuring $CaCl_2$ content in solution, drained solution from Tofu and in agar-agar gel ana Tofu. Empirical linear correlation equations between conductivity (Y, mho) and calcium content (C, Mole $\bar{C}$, mg% wet basis) were obtained for the quick estimation of calcium content during Tofu processings. Equations with plate II type electrode were Y=0.6364C+0.0775 for drained solution with r=0.99, and $Y=7.1503{\times}10^{-5}\bar{C}-2.9895{\times}10^{-3}$ for Tofu with r=0.91,respectively.

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