PURIFICATION OF THE MULTIENZYME COMPLEX FOR FATTY-ACID OXIDATION FROM PSEUDOMONAS-FRAGI AND RECONSTITUTION OF THE FATTY-ACID OXIDATION SYSTEM

被引:48
作者
IMAMURA, S
UEDA, S
MIZUGAKI, M
KAWAGUCHI, A
机构
[1] UNIV TOKYO, COLL ARTS & SCI, DEPT BIOL, TOKYO 153, JAPAN
[2] TOHOKU UNIV HOSP, DEPT PHARMACEUT SCI, SENDAI, MIYAGI 980, JAPAN
[3] TOYO JOZO CO LTD, OHITO, SHIZUOKA 41023, JAPAN
关键词
D O I
10.1093/oxfordjournals.jbchem.a123023
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The multienzyme complex for fatty acid oxidation was purified from Pseudomonas fragi, which was grown on oleic acid as the sole carbon source. This complex exhibited enoyl-CoA hydratase [EC 4.2.1.17], 3-hydroxyacyl-CoA dehydrogenase [EC 1.1.1.35], 3-oxoacyl-CoA thiolase [EC 2.3.1.16], cis-3, trans-2-enoyl-CoA isomerase [EC 5.3.3.3], and 3-hydroxy-acyl-CoA epimerase [EC 5.1.2.3] activities. The molecular weight of the native complex was estimated to be 240,000. Two types of subunits, with molecular weights of 73,000 and 42,000, were identified. The complex was composed of two copies each of the 73,000- and 42,000-Da subunits. The β-oxidation system was reconstituted in vitro using the multienzyme complex, acyl-CoA synthetase and acyl-CoA oxidase. This reconstituted system completely oxidized saturated fatty acids with acyl chains of from 4 to 18 carbon atoms as well as unsaturated fatty acids having cis double bonds extending from odd-numbered carbon atoms. However, unsaturated fatty acids having cis double bonds extending from even-numbered carbon atoms were not completely oxidized to acetyl-CoA: about 5mol of acetyl-CoA was produced from 1 mol of linoleic or a-linolenic acid, and about 2 mol of acetyl-CoA from 1 mol of γ-linolenic acid. These results suggested that the 3-hydroxyacyl-CoA epimerase in the complex was not operative. When the epimerase was by-passed by the addition of 2,4-dienoyl-CoA reductase to the reconstituted system, unsaturated fatty acids with cis double bonds extending from even-numbered carbon atoms were also completely degraded to acetyl-CoA. © 1990 COPYRIGHT, 1990 BY THE JOURNAL OF BIOCHEMISTRY.
引用
收藏
页码:184 / 189
页数:6
相关论文
共 19 条
[1]  
CUEBAS D, 1982, J BIOL CHEM, V257, P4140
[2]  
DOMMES P, 1983, J BIOL CHEM, V258, P846
[3]  
DOMMES V, 1984, J BIOL CHEM, V259, P1781
[4]  
DOMMES V, 1981, J BIOL CHEM, V256, P8259
[5]   PURIFICATION BY AFFINITY-CHROMATOGRAPHY OF 2,4-DIENOYL-COA REDUCTASES FROM BOVINE LIVER AND ESCHERICHIA-COLI [J].
DOMMES, V ;
LUSTER, W ;
CVETANOVIC, M ;
KUNAU, WH .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1982, 125 (02) :335-341
[6]   SIMPLE METHOD FOR QUANTITIVE DENSITOMETRY OF POLYACRYLAMIDE GELS USING FAST GREEN [J].
GOROVSKY, MA ;
CARLSON, K ;
ROSENBAUM, JL .
ANALYTICAL BIOCHEMISTRY, 1970, 35 (02) :359-+
[7]   BETA-OXIDATION OF POLY-UNSATURATED FATTY-ACIDS HAVING DOUBLE-BONDS AT EVEN-NUMBERED POSITIONS IN ISOLATED RAT-LIVER MITOCHONDRIA [J].
HILTUNEN, JK ;
OSMUNDSEN, H ;
BREMER, J .
BIOCHIMICA ET BIOPHYSICA ACTA, 1983, 752 (02) :223-232
[8]   STUDIES ON THE METABOLISM OF UNSATURATED FATTY-ACIDS .15. PURIFICATION AND PROPERTIES OF 2,4-DIENOYL-COA REDUCTASE FROM RAT-LIVER PEROXISOMES [J].
KIMURA, C ;
KONDO, A ;
KOEDA, N ;
YAMANAKA, H ;
MIZUGAKI, M .
JOURNAL OF BIOCHEMISTRY, 1984, 96 (05) :1463-1469
[9]   DEGRADATION OF UNSATURATED FATTY-ACIDS - IDENTIFICATION OF INTERMEDIATES IN DEGRADATION OF CIS-4-DECENOYL-COA BY EXTRACTS OF BEEF-LIVER MITOCHONDRIA [J].
KUNAU, WH ;
DOMMES, P .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1978, 91 (02) :533-544
[10]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+