Purification of phospholipase B from Penicillium notatum by hydrophobic chromatography on palmitoyl cellulose.
Phospholipase B (lysolecithin acyl-hydrolase, EC 3.1.1.5) from the mycelia of Penicillium notatum (Institute for Fermentation, Osaka, Japan; No.4640) was adsorbed from a crude solution to palmitoyl cellulose. Adsorption was efficient at pH 4 at low ionic strength (10 mM buffer), and at pH 4-9 at hig...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
1980-02-01
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| Series: | Journal of Lipid Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520398230 |
| _version_ | 1818671931478507520 |
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| author | S Imamura Y Horiuti |
| author_facet | S Imamura Y Horiuti |
| author_sort | S Imamura |
| collection | DOAJ |
| description | Phospholipase B (lysolecithin acyl-hydrolase, EC 3.1.1.5) from the mycelia of Penicillium notatum (Institute for Fermentation, Osaka, Japan; No.4640) was adsorbed from a crude solution to palmitoyl cellulose. Adsorption was efficient at pH 4 at low ionic strength (10 mM buffer), and at pH 4-9 at higher ionic strength (1-2M NaCl in 10 mM buffer). The adsorbed enzyme was eluted from the cellulose with a suitable detergent, such as Adekatol SO-120, Triton X-100, or deoxycholate. As an application of this procedure, the enzyme was purified from an extract of the mycelia by column chromatography on a palmitoylated textile (palmitoylated gauze) with an overall recovery of 59% and a 38-fold increase in specific activity. By subsequent column chromatographies on Amberlite XAD-2, Sephadex G-100 and G-150, and DEAE-Sephadex A-50, the enzyme was purified about 4,000-fold to a nearly homogeneous state from a mycelium extract with an overall recovery of 37%. |
| first_indexed | 2024-12-17T07:31:51Z |
| format | Article |
| id | doaj.art-7680b7496ce749c18da7efdde6b2a6e4 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-17T07:31:51Z |
| publishDate | 1980-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-7680b7496ce749c18da7efdde6b2a6e42022-12-21T21:58:28ZengElsevierJournal of Lipid Research0022-22751980-02-01212180185Purification of phospholipase B from Penicillium notatum by hydrophobic chromatography on palmitoyl cellulose.S ImamuraY HoriutiPhospholipase B (lysolecithin acyl-hydrolase, EC 3.1.1.5) from the mycelia of Penicillium notatum (Institute for Fermentation, Osaka, Japan; No.4640) was adsorbed from a crude solution to palmitoyl cellulose. Adsorption was efficient at pH 4 at low ionic strength (10 mM buffer), and at pH 4-9 at higher ionic strength (1-2M NaCl in 10 mM buffer). The adsorbed enzyme was eluted from the cellulose with a suitable detergent, such as Adekatol SO-120, Triton X-100, or deoxycholate. As an application of this procedure, the enzyme was purified from an extract of the mycelia by column chromatography on a palmitoylated textile (palmitoylated gauze) with an overall recovery of 59% and a 38-fold increase in specific activity. By subsequent column chromatographies on Amberlite XAD-2, Sephadex G-100 and G-150, and DEAE-Sephadex A-50, the enzyme was purified about 4,000-fold to a nearly homogeneous state from a mycelium extract with an overall recovery of 37%.http://www.sciencedirect.com/science/article/pii/S0022227520398230 |
| spellingShingle | S Imamura Y Horiuti Purification of phospholipase B from Penicillium notatum by hydrophobic chromatography on palmitoyl cellulose. Journal of Lipid Research |
| title | Purification of phospholipase B from Penicillium notatum by hydrophobic chromatography on palmitoyl cellulose. |
| title_full | Purification of phospholipase B from Penicillium notatum by hydrophobic chromatography on palmitoyl cellulose. |
| title_fullStr | Purification of phospholipase B from Penicillium notatum by hydrophobic chromatography on palmitoyl cellulose. |
| title_full_unstemmed | Purification of phospholipase B from Penicillium notatum by hydrophobic chromatography on palmitoyl cellulose. |
| title_short | Purification of phospholipase B from Penicillium notatum by hydrophobic chromatography on palmitoyl cellulose. |
| title_sort | purification of phospholipase b from penicillium notatum by hydrophobic chromatography on palmitoyl cellulose |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520398230 |
| work_keys_str_mv | AT simamura purificationofphospholipasebfrompenicilliumnotatumbyhydrophobicchromatographyonpalmitoylcellulose AT yhoriuti purificationofphospholipasebfrompenicilliumnotatumbyhydrophobicchromatographyonpalmitoylcellulose |