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Characterization of the Central Metabolic Pathways in Thermoanaerobacter sp Strain X514 via Isotopomer-Assisted Metabolite Analysis
Feng, Xueyang1; Mouttaki, Housna2,3; Lin, Lu2,3,4; Huang, Rick1; Wu, Bing1; Hemme, Christopher L.2,3; He, Zhili2,3; Zhang, Baichen5; Hicks, Leslie M.5; Xu, Jian4; Zhou, Jizhong2,3; Tang, Yinjie J.1
2009-08-01
Source PublicationAPPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume75Issue:15Pages:5001-5008
Abstract

Thermoanaerobacter sp. strain X514 has great potential in biotechnology due to its capacity to ferment a range of C(5) and C(6) sugars to ethanol and other metabolites under thermophilic conditions. This study investigated the central metabolism of strain X514 via (13)C-labeled tracer experiments using either glucose or pyruvate as both carbon and energy sources. X514 grew on minimal medium and thus contains complete biosynthesis pathways for all macromolecule building blocks. Based on genome annotation and isotopic analysis of amino acids, three observations can be obtained about the central metabolic pathways in X514. First, the oxidative pentose phosphate pathway in X514 is not functional, and the tricarboxylic acid cycle is incomplete under fermentative growth conditions. Second, X514 contains (Re)-type citrate synthase activity, although no gene homologous to the recently characterized (Re)-type citrate synthase of Clostridium kluyveri was found. Third, the isoleucine in X514 is derived from acetyl coenzyme A and pyruvate via the citramalate pathway rather than being synthesized from threonine via threonine ammonia-lyase. The functionality of the citramalate synthase gene (cimA [Teth514_1204]) has been confirmed by enzymatic activity assays, while the presence of intracellular citramalate has been detected by mass spectrometry. This study demonstrates the merits of combining (13)C-assisted metabolite analysis, enzyme assays, and metabolite detection not only to examine genome sequence annotations but also to discover novel enzyme activities.

SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1128/AEM.00715-09
WOS KeywordESCHERICHIA-COLI ; FLUX ANALYSIS ; ISOLEUCINE BIOSYNTHESIS ; ETHANOL ; BACTERIUM ; BIOFUELS ; SYNTHASE ; GROWTH ; YIELD
Indexed BySCI
Language英语
WOS Research AreaBiotechnology & Applied Microbiology ; Microbiology
WOS SubjectBiotechnology & Applied Microbiology ; Microbiology
WOS IDWOS:000268311600009
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Document Type期刊论文
Identifierhttp://ir.qibebt.ac.cn/handle/337004/6564
Collection单细胞中心组群
Affiliation1.Washington Univ, Dept Energy Environm & Chem Engn, St Louis, MO 63130 USA
2.Univ Oklahoma, Inst Environm Genom, Norman, OK 73019 USA
3.Univ Oklahoma, Dept Bot & Microbiol, Norman, OK 73019 USA
4.Chinese Acad Sci, Qingdao Inst BioEnergy & Bioproc Technol, Qingdao, Peoples R China
5.Donald Danforth Plant Sci Ctr, St Louis, MO 63132 USA
Recommended Citation
GB/T 7714
Feng, Xueyang,Mouttaki, Housna,Lin, Lu,et al. Characterization of the Central Metabolic Pathways in Thermoanaerobacter sp Strain X514 via Isotopomer-Assisted Metabolite Analysis[J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY,2009,75(15):5001-5008.
APA Feng, Xueyang.,Mouttaki, Housna.,Lin, Lu.,Huang, Rick.,Wu, Bing.,...&Tang, Yinjie J..(2009).Characterization of the Central Metabolic Pathways in Thermoanaerobacter sp Strain X514 via Isotopomer-Assisted Metabolite Analysis.APPLIED AND ENVIRONMENTAL MICROBIOLOGY,75(15),5001-5008.
MLA Feng, Xueyang,et al."Characterization of the Central Metabolic Pathways in Thermoanaerobacter sp Strain X514 via Isotopomer-Assisted Metabolite Analysis".APPLIED AND ENVIRONMENTAL MICROBIOLOGY 75.15(2009):5001-5008.
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