Purification of bovine pancreatic phospholipase A2 by an affinity ultrafiltration technique

被引:13
作者
Teke, Mustafa [1 ]
Telefoncu, Azmi [2 ]
机构
[1] Mugla Univ, Sci & Art Fac, Dept Chem, TR-48000 Mugla, Turkey
[2] Ege Univ, Fac Sci, Dept Biochem, TR-35100 Izmir, Turkey
关键词
Phosholipase A(2) purification; Affinity ultrafiltration; Bovine pancreatic phospholipase A(2);
D O I
10.1016/j.seppur.2008.07.019
中图分类号
TQ [化学工业];
学科分类号
0817 ;
摘要
Phospholipase A(2) (PLA(2); EC 3.1.1.4) is a lipolytic enzyme that hydrolyze phosphoglycerides at the acyl ester bond at the sn-2-position into the corresponding lysophospholipid and fatty acid. Mammalian PLA(2) enzymes are subdivided into three groups: low molecular weight Ca2+-dependent enzymes (sPLA(2)), high molecular weight Ca2+-dependent enzymes (cPLA(2)) and the Ca2+-independent isoforms (iPLA(2)). The object of this study is to discover the synthesis of an affinity complex which could be used in an ultrafiltration system for the biospecific affinity isolation of phospholipase A(2) from mixtures containing other proteins. For this purpose, first, the development of a macromolecular water soluble resin was attempted, based on chitosan bearing phosphatidylethanolamine, a phospholipase A(2) substrate. Then, together with ultrafiltration techniques, the macroligand was employed to purify phospholipase A(2) from bovine pancreas. The enzyme was purified 79-fold and had a high activity yield 76.3%. The enzymatic properties obtained and showed nearly similarity in terms of optimum temperature and pH, molecular weight, Ca2+-dependence with bovine and other mammalian pancreatic PLA(2)'S. (c) 2008 Elsevier B.V. All rights reserved.
引用
收藏
页码:716 / 720
页数:5
相关论文
共 41 条
[1]   Phospholipase A(2) - A structural review [J].
Arni, RK ;
Ward, RJ .
TOXICON, 1996, 34 (08) :827-841
[2]   Regulation and inhibition of phospholipase A2 [J].
Balsinde, J ;
Balboa, MA ;
Insel, PA ;
Dennis, EA .
ANNUAL REVIEW OF PHARMACOLOGY AND TOXICOLOGY, 1999, 39 :175-189
[3]   Purification and biochemical characterization of phospholipase A2 from dromedary pancreas [J].
Ben Bacha, Abir ;
Gargouri, Youssef ;
Bezzine, Sofiane ;
Mejdoub, Hafedh .
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 2006, 1760 (08) :1202-1209
[4]   Purification of turkey pancreatic phospholipase A2 [J].
Ben Salah, R ;
Zouari, N ;
Reinbolt, J ;
Mejdoub, H .
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 2003, 67 (10) :2139-2144
[5]   PURIFICATION AND CHARACTERIZATION OF A SOLUBLE PHOSPHOLIPASE-A2 FROM GUINEA-PIG LUNG [J].
BENNETT, CF ;
MCCARTE, A ;
CROOKE, ST .
BIOCHIMICA ET BIOPHYSICA ACTA, 1990, 1047 (03) :271-283
[6]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[7]  
CAO YZ, 1987, J BIOL CHEM, V262, P16927
[8]   Mammalian phospholipases A2:: mediators of inflammation, proliferation and apoptosis [J].
Capper, EA ;
Marshall, LA .
PROGRESS IN LIPID RESEARCH, 2001, 40 (03) :167-197
[9]   EVOLUTIONARY RELATIONSHIPS AND IMPLICATIONS FOR THE REGULATION OF PHOSPHOLIPASE-A2 FROM SNAKE-VENOM TO HUMAN SECRETED FORMS [J].
DAVIDSON, FF ;
DENNIS, EA .
JOURNAL OF MOLECULAR EVOLUTION, 1990, 31 (03) :228-238
[10]   PURIFICATION AND PROPERTIES OF PHOSPHOLIPASE A FROM PORCINE PANCREAS [J].
DEHAAS, GH ;
POSTEMA, NM ;
NIEUWENHUIZEN, W ;
VANDEENE.LL .
BIOCHIMICA ET BIOPHYSICA ACTA, 1968, 159 (01) :103-+