Identification of an archaeal type II isopentenyl diphosphate isomerase in Methanothermobacter thermautotrophicus

被引:35
作者
Barkley, SJ [1 ]
Cornish, RM [1 ]
Poulter, CD [1 ]
机构
[1] Univ Utah, Dept Chem, Salt Lake City, UT 84112 USA
关键词
D O I
10.1128/JB.186.6.1811-1817.2004
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Isopentenyl diphosphate (IPP):dimethylallyl diphosphate isomerase catalyzes the interconversion of the fundamental five-carbon homoallylic and allylic diphosphate building blocks required for biosynthesis of isoprenoid compounds. Two different isomerases have been reported. The type I enzyme, first characterized in the late 1950s, is widely distributed in eukaryota and eubacteria. The type II enzyme was recently discovered in Streptomyces sp. strain CL190. Open reading frame 48 (ORF48) in the archaeon Methanothermobacter thermautotrophicus encodes a putative type II IPP isomerase. A plasmid-encoded copy of the ORF complemented IPP isomerase activity in vivo in Salmonella enterica serovar Typhimurium strain RMC29, which contains chromosomal knockouts in the genes for type I IPP isomerase (idi) and 1-deoxy-D-xylulose 5-phosphate (dxs). The dxs gene was interrupted with a synthetic operon containing the Saccharomyces cerevisiae genes erg8, erg12, and erg19 allowing for the conversion of mevalonic acid to IPP by, the mevalonate pathway. His(6)-tagged M. thermautotrophicus type II IPP isomerase was produced in Escherichia coli and purified by Ni2+ chromatography. The purified protein was characterized by matrix-assisted laser desorption ionization mass spectrometry. The enzyme has optimal activity at 70degreesC and pH 6.5. NADPH, flavin mononucleotide, and Mg2+ are required cofactors. The steady-state kinetic constants for the archaeal type II IPP isomerase from M. thermautotrophicus are as follows: K-m, 64 muM; specific activity, 0.476 mumol mg(-1) min(-1); and k(cat), 1.6 s(-1).
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页码:1811 / 1817
页数:7
相关论文
共 47 条
[1]  
ANDERSON MS, 1989, J BIOL CHEM, V264, P19169
[2]   A SINGLE REGULATORY GENE INTEGRATES CONTROL OF VITAMIN-B12 SYNTHESIS AND PROPANEDIOL DEGRADATION [J].
BOBIK, TA ;
AILION, M ;
ROTH, JR .
JOURNAL OF BACTERIOLOGY, 1992, 174 (07) :2253-2266
[3]  
BUCKINGHAM J, 1998, DICT NATURAL PRODUCT
[4]   PHOSPHORYLATED INTERMEDIATES IN THE SYNTHESIS OF SQUALENE [J].
CHAYKIN, S ;
LAW, J ;
PHILLIPS, AH ;
TCHEN, TT ;
BLOCH, K .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1958, 44 (10) :998-1004
[5]   Role of accurate mass measurement (±10 ppm) in protein identification strategies employing MS or MS MS and database searching [J].
Clauser, KR ;
Baker, P ;
Burlingame, AL .
ANALYTICAL CHEMISTRY, 1999, 71 (14) :2871-2882
[6]  
CORNFORTH J. W., 1959, TETRAHEDRON LETTERS, V19, P29
[7]   Incorporation of 2-C-methyl-D-erythritol, a putative isoprenoid precursor in the mevalonate-independent pathway, into ubiquinone and menaquinone of Escherichia coli [J].
Duvold, T ;
Cali, P ;
Bravo, JM ;
Rohmer, M .
TETRAHEDRON LETTERS, 1997, 38 (35) :6181-6184
[8]   GENETIC-ANALYSIS OF THE ERF REGION OF THE BACTERIOPHAGE-P22 CHROMOSOME [J].
FENTON, AC ;
POTEETE, AR .
VIROLOGY, 1984, 134 (01) :148-160
[9]   Human isopentenyl diphosphate:dimethylallyl diphosphate isomerase: Overproduction, purification, and characterization [J].
Hahn, FM ;
Xuan, JW ;
Chambers, AF ;
Poulter, CD .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1996, 332 (01) :30-34
[10]   Escherichia coli open reading frame 696 Is idi, a nonessential gene encoding isopentenyl diphosphate isomerase [J].
Hahn, FM ;
Hurlburt, AP ;
Poulter, CD .
JOURNAL OF BACTERIOLOGY, 1999, 181 (15) :4499-4504