• Title/Summary/Keyword: superose column

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Characterization of partially purified 8 kDa antigenic protein of Clonorchis sinensis

  • Chung, Young-Bae;Lee, Me-Jeong;Yang, Hyun-Jong;Chung, Byung-Suk;Lee, Shun-Yu;Choi, Min-Ho;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.40 no.2
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    • pp.83-88
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    • 2002
  • The 8 kDa antigenic protein of Clonorchis sinensis was partially purified by ammonium sulfate precipitation and subsequently by a column chromatographic steps. The purified protein was separated into 7 and 8 kDa protein bands through SDS-tricine gel electrophoresis, while the protein was fecund to migrate to a 8 kDa band in 7.5-15% SDS-PAGE. The molecular weight of the antigen was estimated to be 110 kDa by Superose 6 HR 10/30 gel filtration. The purified antigen strongly reacted with the human sera of clonorchiasis. The hyperimmune sera of BALB/c mice immunized against the 8 kDa protein were reacted with both the crude extract and the excretory-secretory product of adult worms, but not with the metacercarial extract. Immunohistochemical staining demonstrated that the protein was distributed to the tegument and subtegumental cells and also to the seminal receptacle. The present findings suggest that the 8 kDa protein is a partition of the multicomplek protein originating from various organs of adult C. sinenis, and that it is composed of several 7 and 8 kDa proteins.

Characterization of a New Type II Restriction Endonuclease Isolated from streptoverticillium olivoverticillatum (Streptoverticillium olivoverticillatum에서 분리한 새로운 Type II 제한효소 SolI의 특성 연구)

  • Hwang, Hye-Yeon;Yim, Jeong-Bin
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.208-214
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    • 1994
  • We screened many species from a wide variety of bacterial genera for a new type II restriction endonuclease. The purification and characterization of SolI from a soil isolate, Streptoverticillium olivoverticillatum are described here. The enzyme turned out to be an isoschizomer of BamHI. It recognized the hexanucleotide sequence of 5'-G$\downarrow$GATCC-3' and cleaved as in dicated by the arrow, generating a 4 base 5' extension. Unlike its isoschizomer, BamHI, the activity was sensitive to dam methylation within the recognition sequence. Following ammonium sulfate fractionation of the crude extract, heparin-agarose and Affi-gel Blue column chromatography were employed to purify the enzyme. SolI required at least 0.2 mM of $MgCl_2$ for the cleavage to occur. The enzyme exhibited its maximal activity in the absence of NaCl, but was inhibited completely in the presence of 120 mM NaCl. The pH and temperature optima for activity were pH 8.6 and $40^{\circ}C$, respectively. The molecular weight of SolI was estimated to be 43,000 Da by Superose-12 gel filtraion chromatography.

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Purification and Properties of Ribosome-inactivating Proteins from the Leaves of $Cucurbita\;moschata\;D_{UCHESNE}$ (호박$(Cucurbita\;moschata\;D_{UCHESNE})$잎에서 리보즘불활성화 단백질의 분리 및 특성)

  • Lee, Si-Myung;Kim, Yeong-Tae;Hwang, Young-Soo;Cho, Kang-Jin
    • Applied Biological Chemistry
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    • v.40 no.5
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    • pp.375-379
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    • 1997
  • Two ribosome-inactivating proteins, PRIP 1 and PRIP 2 have been isolated from the leaves of $Cucurbita\;moschata\;D_{UCHESNE}$. Crude extracts were purified through ammonium sulfate precipitation and column chromatography using DE-52 cellulose, S-Sepharose, FPLC Suprose 12 HR and FPLC Mono-S. The molecular weights of PRIP 1 and PRIP 2 were 31,000 and 30,500, respectively. PRIP 2 was thermostabe and maintained its activity even after the incubation of the protein at $50^{\circ}C$ for 30 min. In a cell free in vitro translation system using rabbit reticulocyte lysate, protein synthesis was inhibited by the addition of PRIP 1 and PRIP 2. The $IC_{50}$ of PRIP 1 and PRIP 2 were 0.82 nM and 0.79 nM, respectively. The comparison of N-terminal amino acid sequences of the PRIP 1 and PRIP 2 with known RIPs revealed that PRIP 1 shows sequence similarity with Luffin B from Luffa cylindrica and Trichokirin from Trichosanthes kirilowii Maximowicz and PRH) 2 has sequence similarity with Momordin II and MAP 30 from Momordica charantia.

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[Kimchi Pill] Preparation of a Kimchi Pill Using Cyclodextrin ([김치환] Cyclodextrin을 이용한 김치환의 제조)

  • Ann Yong-Geun;Lee Kyung-Haeng
    • The Korean Journal of Food And Nutrition
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    • v.18 no.3
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    • pp.207-218
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    • 2005
  • In room temperature, Kimchi becomes acidified and a little decayed, scenting a bad smell, and It couldn't be well kept. But if it should be made into a pill, it could be preserved for a long time for marketing, with nutrition highly concentrated as well as with no scent. Therefore, making Kimchi into a pill needs drying. When dried Kimchi, lactic acid and fragrant ingredient will vanish along with volatilization. The cyclodextrin(CD) as a stabilizer shows that the protecting rate of volatility of lactic acid in Kimchi is higher before than that of after fermentation, and it is higher at the addition $2\%\;than\;of\;1\%$ in case of Kimchi with CD. But it doesn't give much effect on total sugar, reducing sugar, protein and amino acid. Evaporation rate of lactic acid is the least in freeze dry, and natural dry, heat dry come next, respectively. In heat dry, if dried at more than $60^{\circ}C$ for a long time, Kimchi exudes boiling and scorched scent, causing bitter taste. The result of HPLC with superose 12 column at 280nm and 210nm shows that place and amount of main peak is almost the same, but the distribution of other peaks are different, with the revelation of various peaks like peptide and amino acid. The Kimchi pill made by the addition of $1\%$ CD shows that concentration is eight times higher than general Kimchi, total sugar is $14.4\%$, reducing sugar is $8.8\%$, protein is $4.8\%$, amino acid is $2.4\%$, and other contents are $74.4\%$, acidity is 32.8, and pH is 3.5 each. The result of letting 20 people with obesity, 20 patients with constipation have 30 pills(total weight 30g) three times a day for 60 days reveals they lost $2.29\%$ in weight on the average, and 7 among 20 were all relieved in constipation, and 8 responded that they experienced its efficacy.