Analysis of gluconeogenic and anaplerotic enzymes in Campylobacter jejuni: an essential role for phosphoenolpyruvate carboxykinase
Jyoti Velayudhan,David J. Kelly +1 more
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Sequence and biochemical data indicate that the PYC of C. jejuni is a member of the alpha4beta4, acetyl-CoA-independent class of PYCs, with a 65.8 kDa subunit containing the biotin moiety, and it is concluded that PYK may function in the catabolism of unidentified substrates which are metabolized through PEP.Abstract:
Campylobacter jejuni is unable to utilize glucose as a carbon source due to the absence of the key glycolytic enzyme 6-phosphofructokinase. The genome sequence of C. jejuni NCTC 11168 indicates that homologues of all the appropriate enzymes for gluconeogenesis from phosphoenolpyruvate (PEP) are present, in addition to the anaplerotic enzymes pyruvate carboxylase (PYC), phosphoenolpyruvate carboxykinase (PCK) and malic enzyme (MEZ). Surprisingly, a pyruvate kinase (PYK) homologue is also present. To ascertain the role of these enzymes, insertion mutants in pycA, pycB, pyk and mez were generated. However, this could not be achieved for pckA, indicating that PCK is an essential enzyme in C. jejuni. The lack of PEP synthase and pyruvate orthophosphate dikinase activities confirmed a unique role for PCK in PEP synthesis. The pycA mutant was unable to grow in defined medium with pyruvate or lactate as the major carbon source, thus indicating an important role for PYC in anaplerosis. Sequence and biochemical data indicate that the PYC of C. jejuni is a member of the alpha4beta4, acetyl-CoA-independent class of PYCs, with a 65.8 kDa subunit containing the biotin moiety. Whereas growth of the mez mutant was comparable to that of the wild-type, the pyk mutant displayed a decreased growth rate in complex medium. Nevertheless, the mez and pyk mutants were able to grow with pyruvate, lactate or malate as carbon sources in defined medium. PYK was present in cell extracts at a much higher specific activity [>800 nmol x min(-1) x (mg protein)(-1)] than PYC or PCK [<65 nmol x min(-1) x (mg protein)(-1)], was activated by fructose 1,6-bisphosphate and displayed other regulatory properties strongly indicative of a catabolic role. It is concluded that PYK may function in the catabolism of unidentified substrates which are metabolized through PEP. In view of the high K(m) of MEZ for malate (approximately 9 mM) and the lack of a growth phenotype of the mez mutant, MEZ seems to have only a minor anaplerotic role in C. jejuni.read more
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Journal ArticleDOI
The PEP-pyruvate-oxaloacetate node as the switch point for carbon flux distribution in bacteria.
Uwe Sauer,Bernhard J. Eikmanns +1 more
TL;DR: The present knowledge unequivocally reveals the PEP-pyruvate-oxaloacetate nodes of bacteria to be a fascinating target of metabolic engineering in order to achieve optimized metabolite production.
Journal ArticleDOI
Unique Features of a Highly Pathogenic Campylobacter jejuni Strain
Dirk Hofreuter,Jennifer Tsai,Robert O. Watson,Veronica Novik,Bill Altman,Michelle Benitez,Christina Clark,Clotilde Perbost,Thomas Jarvie,Lei Du,Jorge E. Galán +10 more
TL;DR: A number of unique genetic features which may confer specific metabolic and pathogenic properties on this strain are identified and regions of the C. jejuni genome that are hot spots for the integration of horizontally acquired genetic material are identified.
Journal ArticleDOI
NADPH-generating systems in bacteria and archaea
TL;DR: The major canonical and non-canonical reactions involved in the production and regeneration of NADPH in prokaryotes are described, and their key enzymes are discussed and an overview of how different enzymes have been applied to increase NADPH availability and thereby enhance productivity is provided.
AUTHOR'S CORRECTION Unique Features of a Highly Pathogenic Campylobacter jejuni Strain
Dirk Hofreuter,Jennifer Tsai,Robert O. Watson,Veronica Novik,Bill Altman,Michelle Benitez,Christina Clark,Clotilde Perbost,Thomas Jarvie,Lei Du +9 more
TL;DR: A number of unique genetic features which may confer specific metabolic and pathogenic properties on this strain are identified and regions of the C. jejuni genome that are hot spots for the integration of horizontally acquired genetic material are identified.
Journal ArticleDOI
Genome of the epsilonproteobacterial chemolithoautotroph Sulfurimonas denitrificans.
Stefan M. Sievert,Kathleen M. Scott,Martin G. Klotz,Patrick S. G. Chain,Patrick S. G. Chain,Loren Hauser,James Hemp,Michael Hügler,Michael Hügler,Miriam Land,Alla Lapidus,Frank W. Larimer,Susan Lucas,Stephanie Malfatti,Stephanie Malfatti,Folker Meyer,Ian T. Paulsen,Qinghu Ren,Jörg Simon +18 more
TL;DR: The genome of Sulfurimonas denitrificans DSM1251 was sequenced in this article, showing a branched electron transport chain with genes encoding complexes for the oxidation of hydrogen, reduced sulfur compounds, and formate and the reduction of nitrate and oxygen.
References
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Book
Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Book
Bergey's Manual of Systematic Bacteriology
TL;DR: BCL3 and Sheehy cite Bergey's manual of determinative bacteriology of which systematic bacteriology, first edition, is an expansion.
Journal ArticleDOI
A procedure for the isolation of deoxyribonucleic acid from micro-organisms
TL;DR: A method has been described for the isolation of DNA from micro-organisms which yields stable, biologically active, highly polymerized preparations relatively free from protein and RNA, and Representative samples have been characterized for their thermal stability and sedimentation behaviour.
Journal ArticleDOI
The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences
Julian Parkhill,Brendan W. Wren,Karen Mungall,Julian M. Ketley,Carol Churcher,D. Basham,Tracey Chillingworth,Robert L. Davies,Theresa Feltwell,S. Holroyd,Kay Jagels,Andrey V. Karlyshev,Sharon Moule,Mark J. Pallen,Charles W. Penn,Michael A. Quail,Marie-Adèle Rajandream,Kim Rutherford,A. H. M. van Vliet,Sally Whitehead,Bart Barrell +20 more
TL;DR: The genome sequence of C. jejuni NCTC11168 is reported, finding short homopolymeric runs of nucleotides were commonly found in genes encoding the biosynthesis or modification of surface structures, or in closely linked genes of unknown function.
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