Enzymatic determination of phospholipase D activity with choline oxidase

Journal of Biochemistry
S Imamura, Y Horiuti

Abstract

A new enzymatic method was developed for the assay of phospholipase D [phosphatidylcholine phosphatidohydrolase EC 3.1.4.4] from cabbage leaves using choline oxidase from Arthrobacter globiformis cells. The method was based on the estimation of choline by the following series of enzymatic reactions after ending the phospholipase D reaction: Choline + 202 + h2o Choline oxidase Betaine + 2H2O2 2H2O2 + Phenol + 4-Aminoantipyrine Peroxidase Quinoneimine dye + 4H2O The amount of choline was proportional to the amount of resulting quinoneimine dye with an absorbance maximum at 500 nm. The phospholipase D reaction (choline liberation) was carried out at pH 5.5 in the presence of Ca2+ ions and ended by adding EDTA in conc. Tris-HCl buffer, pH 8, to give a final pH of around 8. The initial rate of the phospholipase D reaction was proportional to the enzyme concentration over the absorbance change range of 0 to 0.25 (equivalent to 0-21 micron of choline) under the optimal reaction conditions.

Citations

Oct 8, 2008·Bioscience, Biotechnology, and Biochemistry·Hiroko OtaSusumu Osawa
Sep 19, 2009·Molecular Biology Reports·Bin LiuYongjiong Jia
Aug 10, 2019·Protein Engineering, Design & Selection : PEDS·Jasmina DamnjanovićYugo Iwasaki
Apr 1, 1996·Magnetic Resonance in Medicine : Official Journal of the Society of Magnetic Resonance in Medicine·H Gillham, K M Brindle
Nov 29, 2008·Biotechnology Letters·Jaya Ram SimkhadaJin Cheol Yoo
Aug 2, 2011·The Journal of Biological Chemistry·Evgeny Bulat, Teresa A Garrett
Jul 22, 2018·The Journal of Biological Chemistry·Fernando Salgado-PoloAnastassis Perrakis
Sep 25, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Kristina KraljićMilan Mesić
May 17, 2006·The Protein Journal·Yozo NakazawaKatsumi Takano
Aug 17, 2011·Analytical and Bioanalytical Chemistry·Sara ForneraPeter Walde
Mar 7, 2014·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Matthew G K BeneschDavid N Brindley
May 7, 2013·Expert Opinion on Therapeutic Patents·Efrosini BarbayianniGeorge Kokotos
Apr 28, 2017·Expert Opinion on Therapeutic Patents·Aikaterini NikolaouGeorge Kokotos
Mar 18, 2004·International Journal of Cancer. Journal International Du Cancer·Astrid KehlenCuong Hoang-Vu
Nov 28, 2007·Archives of Pharmacal Research·Jaya Ram SimkhadaJin Cheol Yoo
Nov 10, 2009·Archives of Pharmacal Research·Poonam ManderJin Cheol Yoo
Sep 27, 2008·The American Journal of Pathology·Tae NakasakiMasayuki Miyasaka
Nov 6, 2009·Journal of the American Chemical Society·Sheereen MajdMichael Mayer
Nov 5, 1997·Analytical Biochemistry·A J MorrisJ Engebrecht
May 15, 2021·Critical Reviews in Biotechnology·Peng ZhangJin-Song Shi
Mar 29, 1996·Chemistry and Physics of Lipids·A AbousalhamR Verger
Jan 1, 1989·Toxicon : Official Journal of the International Society on Toxinology·K M Crowell, F Lutz
Aug 20, 1993·Biochimica Et Biophysica Acta·A AbousalhamR Verger
Oct 1, 1991·Journal of Biochemical and Biophysical Methods·J F Takrama, K E Taylor
Jan 1, 1982·The International Journal of Biochemistry·M FujiiK Koga
Jan 1, 1981·Analytical Biochemistry·G M CarmanG Maerker
Feb 18, 2012·Chemical Reviews·Harald M H G Albers, Huib Ovaa
Dec 17, 2016·Journal of Medicinal Chemistry·Lisa M MillerAllan J B Watson
Mar 19, 2005·Langmuir : the ACS Journal of Surfaces and Colloids·Sosaku Ichikawa, Peter Walde

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