PMID: 41998Nov 1, 1979

Isolation and characterization of phospholipase D from fababeans

Lipids
A S AtwalH M Henderson

Abstract

An enzyme activity in crude extract of fababeans hydrolyzed phosphatidylcholine-U-14C to produce choline and phosphatidic acid. This enzyme, phospholipase D, was stable at 50 C in the presence of 5 mM DTT but was inactivated at 55 C. The enzyme was precipitated with cold acetone, concentrated between 30% saturation to 40% saturation with ammonium sulphate, absorbed on calcium phosphate gel and eluted with 0.2 M phosphate buffer. This procedure resulted in a 20-fold increase in specific activity. The activity of fababean phospholipase D was much higher when assayed at 38 C than that at room temperature. There was an obligatory requirement for calcium, and for maximal activity 40 mM calcium was required. A narrow pH optimum of about pH 5.7 was observed. The enzyme activity was extremely dependent on substrate dispersion. When 5 mM phosphatidylcholine (PC) was sonicated with increasing levels of sodium dodecyl sulphate (1 mM to 4 mM), the enzyme activity kept increasing. By using equimolar concentrations of PC and sodium dodecyl sulphate (1 mM to 5 mM), the Michaelis constant (Km) was estimated to be 1.74 mM. Addition of choline and serine at 10 mM concentration reduced phospholipase D activity by 31% and 22%, respectively.

References

Jan 1, 1978·Advances in Lipid Research·M Heller
Sep 1, 1954·Canadian Journal of Biochemistry and Physiology·M KATES
May 1, 1963·Proceedings of the Society for Experimental Biology and Medicine·W M DOIZAKI, L ZIEVE

Citations

Jul 30, 1999·Biochimica Et Biophysica Acta·M Waite
Apr 1, 1977·CRC Critical Reviews in Food Science and Nutrition·N A EskinA Pinsky
Mar 10, 2001·Protein Expression and Purification·S Sharma, M N Gupta

Related Concepts

Drug Stability
Hydrogen-Ion Concentration
Tachigalia
Phosphatidylcholines
Phospholipase D
Phospholipase
Pharmaceutical Plants

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