Characterization of crude lipoxygenase extract from green pea using a modified spectrophotometric method

被引:48
作者
Gökmen, V [1 ]
Bahçeci, S [1 ]
Acar, J [1 ]
机构
[1] Hacettepe Univ, Dept Food Engn, TR-06532 Ankara, Turkey
关键词
lipoxygenase; linoleic acid; green pea; modified spectrophotometric assay; pH and temperature optima;
D O I
10.1007/s00217-002-0518-x
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
In the spectrophotometric assay of lipoxygenase (LOX), the buffered linoleic acid solution used as the reaction medium is not optically clear enough at neutral or lower pH values, due to its limited solubility, to provide a precise, accurate and reproducible estimation of activity as the increase in absorbance at 234 nm. Therefore, an optically clear solution was obtained by formation of the Na-salt of unreacted linoleic acid before absorbance measurement. The modified method was then used to characterize crude LOX from green peas in terms of pH and temperature optima, thermal stability, and kinetic parameters. The optimum pH and temperature for the activity of LOX from green peas were determined to be 6.0 and 30 degreesC, respectively. LOX was found to be very stable at 60 degreesC, but much less stable at 65 degreesC or higher temperatures. The Michaelis constant (Km) and maximum rate (V-max) for linoleic acid were calculated to be 1666 units per mg protein per min and 2.33 mM, respectively.
引用
收藏
页码:42 / 45
页数:4
相关论文
共 38 条
[1]   INHIBITION OF GREEN BEAN LIPOXYGENASE BY CYANIDE [J].
ADAMS, JB .
FOOD CHEMISTRY, 1989, 31 (04) :243-250
[2]   PROPERTIES OF BROAD BEAN LIPOXYGENASE [J].
ALOBAIDY, HM ;
SIDDIQI, AM .
JOURNAL OF FOOD SCIENCE, 1981, 46 (02) :622-&
[3]  
ALSOE KO, 1988, LEBENSM WISS TECHNOL, V21, P8
[4]   THE EFFECT OF LIPID-DEGRADING ENZYME-ACTIVITIES ON QUALITY OF BLANCHED AND UNBLANCHED FROZEN STORED CAULIFLOWER ESTIMATED BY SENSORY AND INSTRUMENTAL ANALYSIS [J].
BAARDSETH, P ;
NAESSET, E .
FOOD CHEMISTRY, 1989, 32 (01) :39-46
[5]   Characterization of hydroperoxides and carbonyl compounds obtained by lipoxygenase extracts of selected microorganisms [J].
Bisakowski, B ;
Perraud, X ;
Kermasha, S .
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 1997, 61 (08) :1262-1269
[6]   Characterization of purified lipoxygenase extracts from Fusarium proliferatum [J].
Bisakowski, B ;
Kermasha, S ;
Spinnler, E .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 1998, 46 (06) :2382-2388
[7]   KIWIFRUIT LIPOXYGENASE - PREPARATION AND CHARACTERISTICS [J].
BOYES, S ;
PERERA, C ;
YOUNG, H .
JOURNAL OF FOOD SCIENCE, 1992, 57 (06) :1390-1394
[8]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[9]   Kinetics of thermal inactivation of pea seed lipoxygenases and the effect of additives on their thermostability [J].
Busto, MD ;
Apenten, RKO ;
Robinson, DS ;
Wu, Z ;
Casey, R ;
Hughes, RK .
FOOD CHEMISTRY, 1999, 65 (03) :323-329
[10]   PURIFICATION AND CHARACTERIZATION OF A LIPOXYGENASE FROM IMMATURE ENGLISH PEAS [J].
CHEN, AO ;
WHITAKER, JR .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 1986, 34 (02) :203-211