Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Hydrolytic and transphosphatidylation activities of phospholipase D from Savoy cabbage towards lysophosphatidylcholine

Virto, Carmen LU ; Svensson, Ingemar LU and Adlercreutz, Patrick LU orcid (2000) In Chemistry and Physics of Lipids 106(1). p.41-51
Abstract

The hydrolysis and transphosphatidylation of lysophosphatidylcholine (LPC), with a partially purified preparation of phospholipase D (PL D) from Savoy cabbage, was investigated. These reactions were about 20 times slower than the hydrolysis of phosphatidylcholine (PC) in a micellar system. For the transfer reaction, 2 M glycerol was included in the media, which suppressed the hydrolytic reaction. Both reactions presented similar V(max) values, suggesting that the formation of the phosphatidyl-enzyme intermediate is the rate-limiting step. The enzyme had an absolute requirement for Ca2+, and the optimum concentration was approximately 40 mM CaCl2. K(Ca)(app) was calculated to be 8.6±0.74 mM for the hydrolytic and... (More)

The hydrolysis and transphosphatidylation of lysophosphatidylcholine (LPC), with a partially purified preparation of phospholipase D (PL D) from Savoy cabbage, was investigated. These reactions were about 20 times slower than the hydrolysis of phosphatidylcholine (PC) in a micellar system. For the transfer reaction, 2 M glycerol was included in the media, which suppressed the hydrolytic reaction. Both reactions presented similar V(max) values, suggesting that the formation of the phosphatidyl-enzyme intermediate is the rate-limiting step. The enzyme had an absolute requirement for Ca2+, and the optimum concentration was approximately 40 mM CaCl2. K(Ca)(app) was calculated to be 8.6±0.74 mM for the hydrolytic and 10±0.97 mM for the transphosphatidylation reaction. Both activities reached a maximum at pH 5.5, independent of Ca2+ concentration. Kinetic studies showed that the Km(app) for the glycerol in the transphosphatidylation reaction is 388±37 mM. Km(app) for the lysophosphatidylcholine depended on Ca2+ concentration and fell between 1 and 3 mM at CaCl2 concentrations from 4 to 40 mM. SDS, TX-100, and CTAB did not activate the enzyme as reported for phosphatidylcholine hydrolysis; on the contrary, reaction rates decreased at detergent concentrations at or above that of lysophosphatidylcholine. Copyright (C) 2000 Elsevier Science Ireland Ltd.

(Less)
Please use this url to cite or link to this publication:
author
; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Hydrolysis, Lysophosphatidic acid, Lysophosphatidylcholine, Lysophosphatidylglycerol, Phospholipase D, Transphosphatidylation
in
Chemistry and Physics of Lipids
volume
106
issue
1
pages
11 pages
publisher
Elsevier
external identifiers
  • scopus:0034213456
  • pmid:10878234
ISSN
0009-3084
DOI
10.1016/S0009-3084(00)00130-4
language
English
LU publication?
yes
id
f31915c6-ee6d-4368-a468-d75fe17a24a3
date added to LUP
2019-06-20 15:53:50
date last changed
2025-04-04 14:53:01
@article{f31915c6-ee6d-4368-a468-d75fe17a24a3,
  abstract     = {{<p>The hydrolysis and transphosphatidylation of lysophosphatidylcholine (LPC), with a partially purified preparation of phospholipase D (PL D) from Savoy cabbage, was investigated. These reactions were about 20 times slower than the hydrolysis of phosphatidylcholine (PC) in a micellar system. For the transfer reaction, 2 M glycerol was included in the media, which suppressed the hydrolytic reaction. Both reactions presented similar V(max) values, suggesting that the formation of the phosphatidyl-enzyme intermediate is the rate-limiting step. The enzyme had an absolute requirement for Ca<sup>2+</sup>, and the optimum concentration was approximately 40 mM CaCl<sub>2</sub>. K(Ca)(app) was calculated to be 8.6±0.74 mM for the hydrolytic and 10±0.97 mM for the transphosphatidylation reaction. Both activities reached a maximum at pH 5.5, independent of Ca<sup>2+</sup> concentration. Kinetic studies showed that the Km(app) for the glycerol in the transphosphatidylation reaction is 388±37 mM. Km(app) for the lysophosphatidylcholine depended on Ca<sup>2+</sup> concentration and fell between 1 and 3 mM at CaCl<sub>2</sub> concentrations from 4 to 40 mM. SDS, TX-100, and CTAB did not activate the enzyme as reported for phosphatidylcholine hydrolysis; on the contrary, reaction rates decreased at detergent concentrations at or above that of lysophosphatidylcholine. Copyright (C) 2000 Elsevier Science Ireland Ltd.</p>}},
  author       = {{Virto, Carmen and Svensson, Ingemar and Adlercreutz, Patrick}},
  issn         = {{0009-3084}},
  keywords     = {{Hydrolysis; Lysophosphatidic acid; Lysophosphatidylcholine; Lysophosphatidylglycerol; Phospholipase D; Transphosphatidylation}},
  language     = {{eng}},
  month        = {{06}},
  number       = {{1}},
  pages        = {{41--51}},
  publisher    = {{Elsevier}},
  series       = {{Chemistry and Physics of Lipids}},
  title        = {{Hydrolytic and transphosphatidylation activities of phospholipase D from Savoy cabbage towards lysophosphatidylcholine}},
  url          = {{http://dx.doi.org/10.1016/S0009-3084(00)00130-4}},
  doi          = {{10.1016/S0009-3084(00)00130-4}},
  volume       = {{106}},
  year         = {{2000}},
}