Purification of phospholipase C from Bacillus cereus by hydrophobic chromatography on palmitoyl cellulose.
Phospholipase C (phosphatidylcholine choline-phosphohydrolase, EC 3.1.4.E) from Bacillus cereus (IAM-1208) was adsorbed to palmitoyl cellulose from a crude enzyme solution at pH 5–9. The adsorption was not influenced by ionic strength up to 2 M NaCl. The adsorbed enzyme was eluted almost completely...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
1979-05-01
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| Series: | Journal of Lipid Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520406078 |
| _version_ | 1818647504480108544 |
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| author | S Imamura Y Horiuti |
| author_facet | S Imamura Y Horiuti |
| author_sort | S Imamura |
| collection | DOAJ |
| description | Phospholipase C (phosphatidylcholine choline-phosphohydrolase, EC 3.1.4.E) from Bacillus cereus (IAM-1208) was adsorbed to palmitoyl cellulose from a crude enzyme solution at pH 5–9. The adsorption was not influenced by ionic strength up to 2 M NaCl. The adsorbed enzyme was eluted almost completely by washing the cellulose with a suitable detergent, such as Triton X-100, Adekatol SO-120, Cation DT-205, or sodium deoxycholate. The enzyme was then purified by column chromatography on a palmitoylated textile (palmitoylated gauze) with an overall recovery of 91% and a 467-fold increase in specific activity over that of enzyme in the crude culture supernatant. Subsequent fractionation with acetone and chromatography on a Sephadex G-75 column separated two nearly homogeneous phospholipase C's. The enzyme adsorbed on palmitoyl cellulose was active, although its activity was about one-fourth that of free phospholipase C. Therefore, the enzyme appeared to be adsorbed to the cellulose through a hydrophobic site that was distinct from the catalytic site on the enzyme molecule. |
| first_indexed | 2024-12-17T01:03:35Z |
| format | Article |
| id | doaj.art-f368bfaa7b864f0aa604e16d0ef53cae |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-17T01:03:35Z |
| publishDate | 1979-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-f368bfaa7b864f0aa604e16d0ef53cae2022-12-21T22:09:21ZengElsevierJournal of Lipid Research0022-22751979-05-01204519524Purification of phospholipase C from Bacillus cereus by hydrophobic chromatography on palmitoyl cellulose.S ImamuraY HoriutiPhospholipase C (phosphatidylcholine choline-phosphohydrolase, EC 3.1.4.E) from Bacillus cereus (IAM-1208) was adsorbed to palmitoyl cellulose from a crude enzyme solution at pH 5–9. The adsorption was not influenced by ionic strength up to 2 M NaCl. The adsorbed enzyme was eluted almost completely by washing the cellulose with a suitable detergent, such as Triton X-100, Adekatol SO-120, Cation DT-205, or sodium deoxycholate. The enzyme was then purified by column chromatography on a palmitoylated textile (palmitoylated gauze) with an overall recovery of 91% and a 467-fold increase in specific activity over that of enzyme in the crude culture supernatant. Subsequent fractionation with acetone and chromatography on a Sephadex G-75 column separated two nearly homogeneous phospholipase C's. The enzyme adsorbed on palmitoyl cellulose was active, although its activity was about one-fourth that of free phospholipase C. Therefore, the enzyme appeared to be adsorbed to the cellulose through a hydrophobic site that was distinct from the catalytic site on the enzyme molecule.http://www.sciencedirect.com/science/article/pii/S0022227520406078 |
| spellingShingle | S Imamura Y Horiuti Purification of phospholipase C from Bacillus cereus by hydrophobic chromatography on palmitoyl cellulose. Journal of Lipid Research |
| title | Purification of phospholipase C from Bacillus cereus by hydrophobic chromatography on palmitoyl cellulose. |
| title_full | Purification of phospholipase C from Bacillus cereus by hydrophobic chromatography on palmitoyl cellulose. |
| title_fullStr | Purification of phospholipase C from Bacillus cereus by hydrophobic chromatography on palmitoyl cellulose. |
| title_full_unstemmed | Purification of phospholipase C from Bacillus cereus by hydrophobic chromatography on palmitoyl cellulose. |
| title_short | Purification of phospholipase C from Bacillus cereus by hydrophobic chromatography on palmitoyl cellulose. |
| title_sort | purification of phospholipase c from bacillus cereus by hydrophobic chromatography on palmitoyl cellulose |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520406078 |
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