The release of heparinase from the periplasmic space of Flavobacterium heparinum by three-step osmotic shock
Heparinase was released from the periplasmic space of Flavobacterium heparinum by three-step osmotic shock procedure. The procedure involves resuspending exponentially growing cells consecutively into (1) 40% sucrose, (2) 10 mM sodium phosphate, 2 mM magnesium chloride, pH 7, and (3) 10 mM sodium ph...
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| Veröffentlicht in: | Applied biochemistry and biotechnology 1991-08, Vol.30 (2), p.137-148 |
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| container_title | Applied biochemistry and biotechnology |
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| creator | ZIMMERMANN, J. J. F ODDIE, K LANGER, R COONEY, C. L |
| description | Heparinase was released from the periplasmic space of Flavobacterium heparinum by three-step osmotic shock procedure. The procedure involves resuspending exponentially growing cells consecutively into (1) 40% sucrose, (2) 10 mM sodium phosphate, 2 mM magnesium chloride, pH 7, and (3) 10 mM sodium phosphate, 300 mM sodium chloride, 2 mM magnesium chloride, pH 7. Typically, 50-75% of the total heparinase activity is recovered by this procedure with an observed 7-15-fold increase in purity. The majority of heparinase activity is released in the final step of the procedure allowing for resolution from cytoplasmic and nonspecific periplasmic material. F. heparinum cells can be stored in 40% sucrose at 4 degrees C for up to one week without significant losses in recovery yields. |
| doi_str_mv | 10.1007/BF02921681 |
| format | Article |
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F. heparinum cells can be stored in 40% sucrose at 4 degrees C for up to one week without significant losses in recovery yields.</description><identifier>ISSN: 0273-2289</identifier><identifier>EISSN: 1559-0291</identifier><identifier>DOI: 10.1007/BF02921681</identifier><identifier>PMID: 1952929</identifier><identifier>CODEN: ABIBDL</identifier><language>eng</language><publisher>Heidelberg: Springer</publisher><subject>Biological and medical sciences ; Biotechnology ; Cell Membrane - enzymology ; Edetic Acid ; Enzyme engineering ; Flavobacterium - enzymology ; Flavobacterium - growth & development ; Fundamental and applied biological sciences. Psychology ; Heparin Lyase ; heparinase ; Hydrogen-Ion Concentration ; Improved methods for extraction and purification of enzymes ; Magnesium ; Methods. Procedures. 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J. F</creatorcontrib><creatorcontrib>ODDIE, K</creatorcontrib><creatorcontrib>LANGER, R</creatorcontrib><creatorcontrib>COONEY, C. L</creatorcontrib><title>The release of heparinase from the periplasmic space of Flavobacterium heparinum by three-step osmotic shock</title><title>Applied biochemistry and biotechnology</title><addtitle>Appl Biochem Biotechnol</addtitle><description>Heparinase was released from the periplasmic space of Flavobacterium heparinum by three-step osmotic shock procedure. The procedure involves resuspending exponentially growing cells consecutively into (1) 40% sucrose, (2) 10 mM sodium phosphate, 2 mM magnesium chloride, pH 7, and (3) 10 mM sodium phosphate, 300 mM sodium chloride, 2 mM magnesium chloride, pH 7. Typically, 50-75% of the total heparinase activity is recovered by this procedure with an observed 7-15-fold increase in purity. The majority of heparinase activity is released in the final step of the procedure allowing for resolution from cytoplasmic and nonspecific periplasmic material. F. heparinum cells can be stored in 40% sucrose at 4 degrees C for up to one week without significant losses in recovery yields.</description><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cell Membrane - enzymology</subject><subject>Edetic Acid</subject><subject>Enzyme engineering</subject><subject>Flavobacterium - enzymology</subject><subject>Flavobacterium - growth & development</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Heparin Lyase</subject><subject>heparinase</subject><subject>Hydrogen-Ion Concentration</subject><subject>Improved methods for extraction and purification of enzymes</subject><subject>Magnesium</subject><subject>Methods. Procedures. 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Technologies</topic><topic>Osmolar Concentration</topic><topic>Osmotic Pressure</topic><topic>osmotic shock</topic><topic>periplasmic space</topic><topic>Polysaccharide-Lyases - metabolism</topic><topic>Sucrose</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>ZIMMERMANN, J. J. F</creatorcontrib><creatorcontrib>ODDIE, K</creatorcontrib><creatorcontrib>LANGER, R</creatorcontrib><creatorcontrib>COONEY, C. 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L</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The release of heparinase from the periplasmic space of Flavobacterium heparinum by three-step osmotic shock</atitle><jtitle>Applied biochemistry and biotechnology</jtitle><addtitle>Appl Biochem Biotechnol</addtitle><date>1991-08</date><risdate>1991</risdate><volume>30</volume><issue>2</issue><spage>137</spage><epage>148</epage><pages>137-148</pages><issn>0273-2289</issn><eissn>1559-0291</eissn><coden>ABIBDL</coden><abstract>Heparinase was released from the periplasmic space of Flavobacterium heparinum by three-step osmotic shock procedure. The procedure involves resuspending exponentially growing cells consecutively into (1) 40% sucrose, (2) 10 mM sodium phosphate, 2 mM magnesium chloride, pH 7, and (3) 10 mM sodium phosphate, 300 mM sodium chloride, 2 mM magnesium chloride, pH 7. Typically, 50-75% of the total heparinase activity is recovered by this procedure with an observed 7-15-fold increase in purity. The majority of heparinase activity is released in the final step of the procedure allowing for resolution from cytoplasmic and nonspecific periplasmic material. F. heparinum cells can be stored in 40% sucrose at 4 degrees C for up to one week without significant losses in recovery yields.</abstract><cop>Heidelberg</cop><pub>Springer</pub><pmid>1952929</pmid><doi>10.1007/BF02921681</doi><tpages>12</tpages></addata></record> |
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| identifier | ISSN: 0273-2289 |
| ispartof | Applied biochemistry and biotechnology, 1991-08, Vol.30 (2), p.137-148 |
| issn | 0273-2289 1559-0291 |
| language | eng |
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| subjects | Biological and medical sciences Biotechnology Cell Membrane - enzymology Edetic Acid Enzyme engineering Flavobacterium - enzymology Flavobacterium - growth & development Fundamental and applied biological sciences. Psychology Heparin Lyase heparinase Hydrogen-Ion Concentration Improved methods for extraction and purification of enzymes Magnesium Methods. Procedures. Technologies Osmolar Concentration Osmotic Pressure osmotic shock periplasmic space Polysaccharide-Lyases - metabolism Sucrose |
| title | The release of heparinase from the periplasmic space of Flavobacterium heparinum by three-step osmotic shock |
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