Comparative study on central metabolic fluxes of Bacillus megaterium strains in continuous culture using 13C labelled substrates

被引:25
作者
Fuerch, Tobias
Hollmann, Rajan
Wittmann, Christoph
Wang, Wei
Deckwer, Wolf-Dieter
机构
[1] Tech Univ Carolo Wilhelmina Braunschweig, HZI Helmholtz Zentrum Infektionsforsch, Inst Biochem Engn TU BCE, D-38124 Braunschweig, Germany
[2] Univ Saarland, Inst Biochem Engn, D-66123 Saarbrucken, Germany
关键词
Bacillus megaterium; metabolic flux analysis; central carbon pathways; futile cycle;
D O I
10.1007/s00449-006-0095-7
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Fluxes of central carbon metabolism [glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid cycle (TCA cycle), biomass formation] were determined for several Bacillus megaterium strains (DSM319, WH320, WH323, MS941) in C- and N-limited chemostat cultures by C-13 labelling experiments. The labelling patterns of proteinogenic amino acids were analysed by GC/MS and there from flux ratios at important nodes within the metabolic network could be calculated. On the basis of a stoichiometric metabolic model flux distributions were estimated for the different B. megaterium strains used at various cultivation conditions. Generally all strains exhibited similar metabolic flux distributions, however, several significant changes were found in (1) the glucose flux entering the PPP via the oxidative branch, (2) the reversibilities within the PPP, (3) the relative fluxes of pyruvate and acetyl-CoA fed to the TCA cycle, (4) the fluxes around the pyruvate node involving a futile cycle.
引用
收藏
页码:47 / 59
页数:13
相关论文
共 25 条
[1]   Metabolic network analysis of Bacillus clausii on minimal and semirich medium using 13C-Labeled glucose [J].
Christiansen, T ;
Christensen, B ;
Nielsen, J .
METABOLIC ENGINEERING, 2002, 4 (02) :159-169
[2]   Bacillus subtilis metabolism and energetics in carbon-limited and excess-carbon chemostat culture [J].
Dauner, M ;
Storni, T ;
Sauer, U .
JOURNAL OF BACTERIOLOGY, 2001, 183 (24) :7308-7317
[3]   Metabolic flux analysis with a comprehensive isotopomer model in Bacillus subtilis [J].
Dauner, M ;
Bailey, JE ;
Sauer, U .
BIOTECHNOLOGY AND BIOENGINEERING, 2001, 76 (02) :144-156
[4]   Stoichiometric growth model for riboflavin-producing Bacillus subtilis [J].
Dauner, M ;
Sauer, U .
BIOTECHNOLOGY AND BIOENGINEERING, 2001, 76 (02) :132-143
[5]  
DIESTERHAFT MD, 1973, J BIOL CHEM, V248, P6062
[6]   Metabolic flux profiling of Escherichia coli mutants in central carbon metabolism using GC-MS [J].
Fischer, E ;
Sauer, U .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 2003, 270 (05) :880-891
[7]   High-throughput metabolic flux analysis based on gas chromatography-mass spectrometry derived 13C constraints [J].
Fischer, E ;
Zamboni, N ;
Sauer, U .
ANALYTICAL BIOCHEMISTRY, 2004, 325 (02) :308-316
[8]   Effect of reversible reactions on isotope label redistribution - Analysis of the pentose phosphate pathway [J].
Follstad, BD ;
Stephanopoulos, G .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1998, 252 (03) :360-371
[9]   Pyruvate formation and suppression in recombinant Bacillus megaterium cultivation [J].
Hollmann, R ;
Deckwer, WD .
JOURNAL OF BIOTECHNOLOGY, 2004, 111 (01) :89-96
[10]   MASS ISOTOPOMER PATTERN AND PRECURSOR PRODUCT RELATIONSHIP [J].
LEE, WNP ;
BERGNER, EA ;
GUO, ZK .
BIOLOGICAL MASS SPECTROMETRY, 1992, 21 (02) :114-122