DOCSLIB.ORG

Escherichia Coli K-12 in Response to Trimethylamine-N-Oxide

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Escherichia Coli K-12 in Response to Trimethylamine-N-Oxide Studying the adaptation of Escherichia coli K-12 in response to trimethylamine-N-oxide A thesis submitted in partial fulfilment of the requirements for the degree of Doctor of Philosophy Department of Molecular Biology and Biotechnology, The University of Sheffield Katie Jane Denby MBiolSci (Hons) University of Sheffield September 2016 I Abstract Escherichia coli is a Gram-negative, metabolically versatile facultative anaerobe, which utilises either fermentation, anaerobic respiration or aerobic respiration for energy generation and growth. Trimethylamine-N-oxide (TMAO) is used by E. coli as an alternative terminal electron acceptor, being reduced to trimethylamine (TMA). Although the regulation and operation of the E. coli TMAO respiratory chain (TorCAD) are well characterised, there is no understanding of the dynamic adaptive processes that occur in E. coli during transition from fermentative to TMAO-respiratory growth. Here, glucose-limited chemostat cultures were used to study these adaptive processes. Analyses of the transcriptional and metabolic changes occurring when E. coli K-12 responds to TMAO revealed a number of unexpected components of the adaptive process. Firstly, it was found that growth on a sub-optimal concentration of TMAO resulted in mixed metabolism, with two distinct sub-populations of cells that either activate or do not activate the transcription of the torCAD operon in response to TMAO. DNA methylation was found to contribute to this regulation in response to low concentrations of TMAO. Secondly, it was found that E. coli possesses TMAO demethylase activity, as both the products of this activity, dimethylamine and formaldehyde and the induction of the frmRAB operon was detected when cells were grown in the presence of TMAO. Work was carried out to characterise the regulator protein, FrmR, of the frmRAB operon. Here, it is shown that the FrmR transcriptional repressor specifically reacts with formaldehyde, resulting in the formation of two inter-molecular methylene bridges between the N-terminal Pro2 and Cys35 residues of adjacent FrmR subunits, leading to alleviation of repression. It was also shown that growth yield was higher in the presence of TMAO under aerobic conditions eventhough TMAO reduction to TMA was not detected. Futhermore, the nitrate response regulator NarL does not directly regulate the torCAD operon. II Acknowledgements Firstly, I would like to thank Professor Jeff Green for all his advice, support and patience throughout the last four years. I would also like to thank Professor Robert Poole, whose advice was greatly valued. Thank you to everyone in F10 (both past and present), for their help and making the workplace a lovely place to be. Special thanks must of course go to Dr Matt Rolfe, for his endless patience whilst teaching me all he knows; a lot of the work done here would not have been possible without his help. Thanks also go to Sue Clark for her assistance with flow cytometry and Dr Christa Walther for her advice regarding fluorescence microscopy. I also have to thank our collaborators at Durham University, for their insights into working with FrmR. Thanks also to all of my friends in Sheffield, within MBB, who over the past eight years have made it a truly excellent place to both work and live. Many thanks go to my family. Mum and Dad, without the encouragement and support you have given me over the years, none of this would have been possible. Laura and Sam, thanks for being genuinely awesome siblings who always provide much laughter and fun. Thanks to you all for showing interest in my work and listening to my incessant science-related ramblings. You are truly the best family anyone could have, for that I am privileged. Lastly, and most importantly, is my husband Rich. You have seen more than anyone else, witnessed the tears; restored the lost documents; provided endless support and at times much needed perspective. Thank you for trying to learn and understand biological terms and principles, despite not having a background in science (being a muggle, as you say), in order to perhaps become able to understand my project. You have picked me up and encouraged me to carry on through the tough times and moments of self-doubt. It goes without saying that completing this thesis would not have been possible without you. III Publications and Presentations Publications Denby, K. J., Rolfe, M. D., Crick, E., Sanguinetti, G., Poole, R. K., and Green, J. (2015) Adaptation of anaerobic cultures of Escherichia coli K-12 in response to environmental trimethylamine-N-oxide. Environmental Microbiology, 17: 2477-2491. Denby, K. J., Iwig, J., Bisson, C., Westwood, J., Rolfe, M. D., Sedelnikova, S. E., Higgins, K., Maroney, M. J., Baker, P. J., Chivers, P. T., and Green, J. (2016). The mechanism of a formaldehyde-sensing transcriptional regulator. Scientific Reports. Submitted. Presentations Society of General Microbiology (SGM) Autumn Conference - University of Sussex (2-4th September 2013) Poster presented: Transcriptomic and biochemical changes in Escherichia coli during a shift from fermentation to Trimethylamine-N-oxide (TMAO) respiration Bacterial Electron Transfer Processes and Their Regulation Conference - Vimeiro, Portugal (15-18th March 2015) Poster presented: Adaptation of anaerobic cultures of Escherichia coli K-12 in response to environmental trimethylamine-N-oxide IV Table of contents Abstract ..................................................................................................................................... I Acknowledgements ................................................................................................................. III Publications and Presentations ............................................................................................... IV List of Figures .......................................................................................................................... IX List of Tables .......................................................................................................................... XII 1. Introduction ......................................................................................................................... 1 1.1. The metabolic modes of Escherichia coli ...................................................................... 1 1.2. The hierarchy of metabolism in E. coli and its regulation ............................................. 3 1.2.1. FNR ......................................................................................................................... 5 1.2.2. The ArcBA system .................................................................................................. 6 1.2.3. The Nar system ...................................................................................................... 9 1.3. Reprogramming of the metabolic mode of E. coli under stable (steady-state) and dynamic conditions (transitions) ....................................................................................... 10 1.3.1. Gene expression and transcription factor (TF) activities under steady-state conditions in response to oxygen availability ................................................................ 10 1.3.2. Gene expression and transcription factor (TF) activities during transitions between oxygen-replete and oxygen-deficient environments ..................................... 12 1.4. Trimethylamine-N-oxide (TMAO) ............................................................................... 14 1.4.1. Bacterial TMAO respiration.................................................................................. 14 1.4.2. TMAO and its role in human disease ................................................................... 26 1.4.3. TMAO and fish spoilage ....................................................................................... 30 1.5. Mechanisms of transcriptional repression ................................................................. 31 1.6. Aims of this work ........................................................................................................ 32 2. Materials and Methods ...................................................................................................... 34 2.1. Strains and plasmids ................................................................................................... 34 2.2. Media .......................................................................................................................... 34 2.2.1. Rich media ............................................................................................................ 34 2.2.2. Minimal media ..................................................................................................... 34 2.2.3. Media supplements ............................................................................................. 37 2.3. Culture techniques ...................................................................................................... 37 2.3.1. Chemostat culture of Escherichia coli MG1655 ................................................... 37 2.3.2. Aerobic batch culture of Escherichia coli MG1655 .............................................. 38 2.3.3. Anaerobic batch culture of Escherichia coli MG1655 .......................................... 38 2.4. Cell viability assays ...................................................................................................... 38 2.5. Formaldehyde-sensitivity of FrmR mutants in vivo ...................................................
Load more

Recommended publications
  • Pneumoniae CG43
    YjcC, a c-di-GMP Phosphodiesterase Protein, Regulates the Oxidative Stress Response and Virulence of Klebsiella pneumoniae CG43 Ching-Jou Huang1, Zhe-Chong Wang2, Hsi-Yuan Huang3, Hsien-Da Huang2,3, Hwei-Ling Peng1,2* 1 Institute of Molecular Medicine and Biological Technology, National Chiao Tung University, Hsin Chu, Taiwan, Republic of China, 2 Department of Biological Science and Technology, National Chiao Tung University, Hsin Chu, Taiwan, Republic of China, 3 Institute of Bioinformatics and Systems Biology, National Chiao Tung University, Hsin Chu, Taiwan, Republic of China Abstract This study shows that the expression of yjcC, an in vivo expression (IVE) gene, and the stress response regulatory genes soxR, soxS, and rpoS are paraquat inducible in Klebsiella pneumoniae CG43. The deletion of rpoS or soxRS decreased yjcC expression, implying an RpoS- or SoxRS-dependent control. After paraquat or H2O2 treatment, the deletion of yjcC reduced bacterial survival. These effects could be complemented by introducing the DyjcC mutant with the YjcC-expression plasmid pJR1. The recombinant protein containing only the YjcC-EAL domain exhibited phosphodiesterase (PDE) activity; overexpression of yjcC has lower levels of cyclic di-GMP. The yjcC deletion mutant also exhibited increased reactive oxygen species (ROS) formation, oxidation damage, and oxidative stress scavenging activity. In addition, the yjcC deletion reduced capsular polysaccharide production in the bacteria, but increased the LD50 in mice, biofilm formation, and type 3 fimbriae major pilin MrkA production. Finally, a comparative transcriptome analysis showed 34 upregulated and 29 downregulated genes with the increased production of YjcC. The activated gene products include glutaredoxin I, thioredoxin, heat shock proteins, chaperone, and MrkHI, and proteins for energy metabolism (transporters, cell surface structure, and transcriptional regulation).
    [Show full text]
  • Download Via Github on RNA-Seq and Chip-Seq Data Analysis and the University of And/Or Bioconductor [33–35, 38–40]
    Kumka and Bauer BMC Genomics (2015) 16:895 DOI 10.1186/s12864-015-2162-4 RESEARCH ARTICLE Open Access Analysis of the FnrL regulon in Rhodobacter capsulatus reveals limited regulon overlap with orthologues from Rhodobacter sphaeroides and Escherichia coli Joseph E. Kumka and Carl E. Bauer* Abstract Background: FNR homologues constitute an important class of transcription factors that control a wide range of anaerobic physiological functions in a number of bacterial species. Since FNR homologues are some of the most pervasive transcription factors, an understanding of their involvement in regulating anaerobic gene expression in different species sheds light on evolutionary similarity and differences. To address this question, we used a combination of high throughput RNA-Seq and ChIP-Seq analysis to define the extent of the FnrL regulon in Rhodobacter capsulatus and related our results to that of FnrL in Rhodobacter sphaeroides and FNR in Escherichia coli. Results: Our RNA-seq results show that FnrL affects the expression of 807 genes, which accounts for over 20 % of the Rba. capsulatus genome. ChIP-seq results indicate that 42 of these genes are directly regulated by FnrL. Importantly, this includes genes involved in the synthesis of the anoxygenic photosystem. Similarly, FnrL in Rba. sphaeroides affects 24 % of its genome, however, only 171 genes are differentially expressed in common between two Rhodobacter species, suggesting significant divergence in regulation. Conclusions: We show that FnrL in Rba. capsulatus activates photosynthesis while in Rba. sphaeroides FnrL regulation reported to involve repression of the photosystem. This analysis highlights important differences in transcriptional control of photosynthetic events and other metabolic processes controlled by FnrL orthologues in closely related Rhodobacter species.
    [Show full text]
  • Identification of Alternative Mitochondrial Electron Transport
    International Journal of Molecular Sciences Article Identification of Alternative Mitochondrial Electron Transport Pathway Components in Chickpea Indicates a Differential Response to Salinity Stress between Cultivars Crystal Sweetman * , Troy K. Miller, Nicholas J. Booth, Yuri Shavrukov , Colin L.D. Jenkins, Kathleen L. Soole and David A. Day College of Science & Engineering, Flinders University, GPO Box 5100, Adelaide SA 5001, Australia; troy.miller@flinders.edu.au (T.K.M.); nick.booth@flinders.edu.au (N.J.B.); yuri.shavrukov@flinders.edu.au (Y.S.); colin.jenkins@flinders.edu.au (C.L.D.J.); kathleen.soole@flinders.edu.au (K.L.S.); david.day@flinders.edu.au (D.A.D.) * Correspondence: crystal.sweetman@flinders.edu.au; Tel.: +61-8-82012790 Received: 25 April 2020; Accepted: 27 May 2020; Published: 28 May 2020 Abstract: All plants contain an alternative electron transport pathway (AP) in their mitochondria, consisting of the alternative oxidase (AOX) and type 2 NAD(P)H dehydrogenase (ND) families, that are thought to play a role in controlling oxidative stress responses at the cellular level. These alternative electron transport components have been extensively studied in plants like Arabidopsis and stress inducible isoforms identified, but we know very little about them in the important crop plant chickpea. Here we identify AP components in chickpea (Cicer arietinum) and explore their response to stress at the transcript level. Based on sequence similarity with the functionally characterized proteins of Arabidopsis thaliana, five putative internal (matrix)-facing NAD(P)H dehydrogenases (CaNDA1-4 and CaNDC1) and four putative external (inter-membrane space)-facing NAD(P)H dehydrogenases (CaNDB1-4) were identified in chickpea.
    [Show full text]
  • The Escherichia Coli Narl Receiver Domain Regulates Transcription
    Katsir et al. BMC Microbiology (2015) 15:174 DOI 10.1186/s12866-015-0502-9 RESEARCH ARTICLE Open Access The Escherichia coli NarL receiver domain regulates transcription through promoter specific functions Galit Katsir1,3,4, Michael Jarvis2,5, Martin Phillips1, Zhongcai Ma2,6 and Robert P. Gunsalus2,3* Abstract Background: The Escherichia coli response regulator NarL controls transcription of genes involved in nitrate respiration during anaerobiosis. NarL consists of two domains joined by a linker that wraps around the interdomain interface. Phosphorylation of the NarL N-terminal receiver domain (RD) releases the, otherwise sequestered, C-terminal output domain (OD) that subsequently binds specific DNA promoter sites to repress or activate gene expression. The aim of this study is to investigate the extent to which the NarL OD and RD function independently to regulate transcription, and the affect of the linker on OD function. Results: NarL OD constructs containing different linker segments were examined for their ability to repress frdA-lacZ or activate narG-lacZ reporter fusion genes. These in vivo expression assays revealed that the NarL OD, in the absence or presence of linker helix α6, constitutively repressed frdA-lacZ expression regardless of nitrate availability. However, the presence of the linker loop α5-α6reversedthisrepressionandalsoshowedimpairedDNAbindingin vitro.The OD alone could not activate narG-lacZ expression; this activity required the presence of the NarL RD. A footprint assay demonstrated that the NarL OD only partially bound recognition sites at the narG promoter, and the binding affinity was increased by the presence of the phosphorylated RD. Analytical ultracentrifugation used to examine domain oligomerization showed that the NarL RD forms dimers in solution while the OD is monomeric.
    [Show full text]
  • Pleuropneumoniae Gene Expression
    Global Effects of Catecholamines on Actinobacillus pleuropneumoniae Gene Expression Lu Li, Zhuofei Xu, Yang Zhou, Lili Sun, Ziduo Liu, Huanchun Chen, Rui Zhou* Division of Animal Infectious Diseases in State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China Abstract Bacteria can use mammalian hormones to modulate pathogenic processes that play essential roles in disease development. Actinobacillus pleuropneumoniae is an important porcine respiratory pathogen causing great economic losses in the pig industry globally. Stress is known to contribute to the outcome of A. pleuropneumoniae infection. To test whether A. pleuropneumoniae could respond to stress hormone catecholamines, gene expression profiles after epinephrine (Epi) and norepinephrine (NE) treatment were compared with those from untreated bacteria. The microarray results showed that 158 and 105 genes were differentially expressed in the presence of Epi and NE, respectively. These genes were assigned to various functional categories including many virulence factors. Only 18 genes were regulated by both hormones. These genes included apxIA (the ApxI toxin structural gene), pgaB (involved in biofilm formation), APL_0443 (an autotransporter adhesin) and genes encoding potential hormone receptors such as tyrP2, the ygiY-ygiX (qseC-qseB) operon and narQ-narP (involved in nitrate metabolism). Further investigations demonstrated that cytotoxic activity was enhanced by Epi but repressed by NE in accordance with apxIA gene expression changes. Biofilm formation was not affected by either of the two hormones despite pgaB expression being affected. Adhesion to host cells was induced by NE but not by Epi, suggesting that the hormones affect other putative adhesins in addition to APL_0443. This study revealed that A.
    [Show full text]
  • Prokaryotic Genome Regulation: a Revolutionary Paradigm
    No. 9] Proc. Jpn. Acad., Ser. B 88 (2012) 485 Review Prokaryotic genome regulation: A revolutionary paradigm † By Akira ISHIHAMA*1, (Communicated by Tasuku HONJO, M.J.A.) Abstract: After determination of the whole genome sequence, the research frontier of bacterial molecular genetics has shifted to reveal the genome regulation under stressful conditions in nature. The gene selectivity of RNA polymerase is modulated after interaction with two groups of regulatory proteins, 7 sigma factors and 300 transcription factors. For identification of regulation targets of transcription factors in Escherichia coli, we have developed Genomic SELEX system and subjected to screening the binding sites of these factors on the genome. The number of regulation targets by a single transcription factor was more than those hitherto recognized, ranging up to hundreds of promoters. The number of transcription factors involved in regulation of a single promoter also increased to as many as 30 regulators. The multi-target transcription factors and the multi-factor promoters were assembled into complex networks of transcription regulation. The most complex network was identified in the regulation cascades of transcription of two master regulators for planktonic growth and biofilm formation. Keywords: transcription regulation, genome regulation, transcription factor, regulation network, genomic SELEX, Escherichia coli developed and employed to reveal the expression of 1. Introduction the whole set of genes on the genome (the tran- In the early stage of molecular biology, Esche- scriptome) under a given culture condition. The high- richia coli served as a model organism of biochemical, throughput microarray has made a break-through biophysical, molecular genetic and biotechnological for providing transcription patterns of the whole set studies.
    [Show full text]
  • Full Text Document (Pdf)
    Kent Academic Repository Full text document (pdf) Citation for published version Dolata, Katarzyna M. and Montero, Isabel Guerrero and Miller, Wayne and Sievers, Susanne and Sura, Thomas and Wolff, Christian and Schlüter, Rabea and Riedel, Katharina and Robinson, Colin (2019) Far-reaching cellular consequences of tat deletion in Escherichia coli revealed by comprehensive proteome analyses. Microbiological Research, 218 . pp. 97-107. ISSN 0944-5013. DOI https://doi.org/10.1016/j.micres.2018.10.008 Link to record in KAR https://kar.kent.ac.uk/70187/ Document Version Publisher pdf Copyright & reuse Content in the Kent Academic Repository is made available for research purposes. Unless otherwise stated all content is protected by copyright and in the absence of an open licence (eg Creative Commons), permissions for further reuse of content should be sought from the publisher, author or other copyright holder. Versions of research The version in the Kent Academic Repository may differ from the final published version. Users are advised to check http://kar.kent.ac.uk for the status of the paper. Users should always cite the published version of record. Enquiries For any further enquiries regarding the licence status of this document, please contact: [email protected] If you believe this document infringes copyright then please contact the KAR admin team with the take-down information provided at http://kar.kent.ac.uk/contact.html Microbiological Research 218 (2019) 97–107 Contents lists available at ScienceDirect Microbiological Research journal homepage: www.elsevier.com/locate/micres Far-reaching cellular consequences of tat deletion in Escherichia coli revealed by comprehensive proteome analyses T Katarzyna M.
    [Show full text]
  • Coupling of Energy to Active Transport of Amino Acids in Escherichia Coli (Mutants/Membrane Vesicles/Ca,Mg-Atpase/Electron Transport/D-Lactate Dehydrogenase) ROBERT D
    Proc. Nat. Acad. Sci. USA Vol. 69, No. 9, pp. 2663-2667, September 1972 Coupling of Energy to Active Transport of Amino Acids in Escherichia coli (mutants/membrane vesicles/Ca,Mg-ATPase/electron transport/D-lactate dehydrogenase) ROBERT D. SIMONI AND MARY K. SHALLENBERGER Department of Biological Sciences, Stanford University, Stanford, California 94305 Communicated by Charles Yanofsky, July 6, 1972 ABSTRACT Active transport of amino acids in isolated *to the lack of oxygen (3). Streptococcus faecalis, an anaerobe membrane vesicles of E. coli ML 308-225 is stimulated by that contains no electron-transport system, can carry out oxidation of D-lactate, and this stimulation is dependent the on electron transport [Kaback, H. R. & Milner, L. S. active membrane transport, presumably using energy- (1970) Proc. Nat. Acad. Sci. USA 66, 1008]. In attempting to yielding reactions of glycolysis (7). It has become essential to relate these results to amino-acid transport in intact relate the observations obtained with isolated vesicles to cells, we isolated mutants of E. coli ML 308-225 that physiological realities of intact cells. We have isolated cells contain defects in D-lactate dehydrogenase (EC 1.1.2.4) in various components involved in and electron transport. Intact cells of these mutants are that contain mutations normal for transport of proline and alanine. We also aerobic metabolism and have tested the effects of these muta- isolated mutants defective in Ca,Mg-stimulated ATPase tions on the ability to perform active transport of amino acids. (EC 3.6.1.3), which is responsible for coupling electron transport to the synthesis of ATP.
    [Show full text]
  • Transcription Regulation of the Expression of the Plasmid Encoded Toxin of Enteroaggregative Escherichia Coli
    TRANSCRIPTION REGULATION OF THE EXPRESSION OF THE PLASMID ENCODED TOXIN OF ENTEROAGGREGATIVE ESCHERICHIA COLI by Jacob Henry Duddy A thesis submitted to The University of Birmingham for the degree of MASTER OF PHILOSOPHY February 2013 School of Biosciences The University of Birmingham University of Birmingham Research Archive e-theses repository This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder. Abstract The pathogenic properties of Enteroaggregative Escherichia coli strain 042 results from the synchronised expression of virulence factors, which include the Plasmid Encoded Toxin. Pet is a member of the serine protease autotransporter of the Enterobacteriaceae family and contributes to infection by cleaving α-fodrin, disrupting the actin cytoskeleton of host cells. The expression of Pet is induced by global transcription factor CRP with further enhancement by the nucleoid associated protein Fis. This study identifies the residues of RNA polymerase, Fis and CRP required for the induction of transcription, thereby clarifying the mechanism of activation employed by the transcription factors. Fis activates transcription from the Fis binding site via a direct interaction with RNA polymerase, facilitated by protein specific determinants. This interaction is dependent on the position of the Fis binding site on the DNA and it subsequent orientation on the helical face of the DNA.
    [Show full text]
  • La Réponse Au NO Au Centre De La Pathogenèse Bactérienne Et Cible D’Antibiotiques Innovants Constance Porrini
    La réponse au NO au centre de la pathogenèse bactérienne et cible d’antibiotiques innovants Constance Porrini To cite this version: Constance Porrini. La réponse au NO au centre de la pathogenèse bactérienne et cible d’antibiotiques innovants. Médecine humaine et pathologie. Université Paris-Saclay, 2020. Français. NNT : 2020UP- ASA008. tel-03175216 HAL Id: tel-03175216 https://pastel.archives-ouvertes.fr/tel-03175216 Submitted on 19 Mar 2021 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. La réponse au NO au centre de la pathogenèse bactérienne et cible d’antibiotiques innovants Thèse de doctorat de l'université Paris-Saclay École doctorale n°581, Agriculture, Alimentation, Biologie, Environnement, Santé - ABIES Spécialité de doctorat : Microbiologie INRAE, AgroParisTech, Micalis Institute, 78350, Jouy-en-Josas, France Référent : AgroParisTech Thèse présentée et soutenue à Paris-Saclay, le 12 juin 2020, par Constance PORRINI Composition du Jury Stéphanie BURY-MONE Présidente Professeure, Université Paris-Saclay Véronique BROUSSOLLE Rapporteur et
    [Show full text]
  • Glycolysis Citric Acid Cycle Oxidative Phosphorylation Calvin Cycle Light
    Stage 3: RuBP regeneration Glycolysis Ribulose 5- Light-Dependent Reaction (Cytosol) phosphate 3 ATP + C6H12O6 + 2 NAD + 2 ADP + 2 Pi 3 ADP + 3 Pi + + 1 GA3P 6 NADP + H Pi NADPH + ADP + Pi ATP 2 C3H4O3 + 2 NADH + 2 H + 2 ATP + 2 H2O 3 CO2 Stage 1: ATP investment ½ glucose + + Glucose 2 H2O 4H + O2 2H Ferredoxin ATP Glyceraldehyde 3- Ribulose 1,5- Light Light Fx iron-sulfur Sakai-Kawada, F Hexokinase phosphate bisphosphate - 4e + center 2016 ADP Calvin Cycle 2H Stroma Mn-Ca cluster + 6 NADP + Light-Independent Reaction Phylloquinone Glucose 6-phosphate + 6 H + 6 Pi Thylakoid Tyr (Stroma) z Fe-S Cyt f Stage 1: carbon membrane Phosphoglucose 6 NADPH P680 P680* PQH fixation 2 Plastocyanin P700 P700* D-(+)-Glucose isomerase Cyt b6 1,3- Pheophytin PQA PQB Fructose 6-phosphate Bisphosphoglycerate ATP Lumen Phosphofructokinase-1 3-Phosphoglycerate ADP Photosystem II P680 2H+ Photosystem I P700 Stage 2: 3-PGA Photosynthesis Fructose 1,6-bisphosphate reduction 2H+ 6 ADP 6 ATP 6 CO2 + 6 H2O C6H12O6 + 6 O2 H+ + 6 Pi Cytochrome b6f Aldolase Plastoquinol-plastocyanin ATP synthase NADH reductase Triose phosphate + + + CO2 + H NAD + CoA-SH isomerase α-Ketoglutarate + Stage 2: 6-carbonTwo 3- NAD+ NADH + H + CO2 Glyceraldehyde 3-phosphate Dihydroxyacetone phosphate carbons Isocitrate α-Ketoglutarate dehydogenase dehydrogenase Glyceraldehyde + Pi + NAD Isocitrate complex 3-phosphate Succinyl CoA Oxidative Phosphorylation dehydrogenase NADH + H+ Electron Transport Chain GDP + Pi 1,3-Bisphosphoglycerate H+ Succinyl CoA GTP + CoA-SH Aconitase synthetase
    [Show full text]
  • Cellular Respiration Liberation of Energy by Oxidation of Food
    Cellular Respiration Liberation of Energy by Oxidation of Food Respiration and Photosynthesis: Photosynthesis uses CO2 and H2O molecules to form C6H12O6 (glucose) and O2. Respiration is just the opposite of photosynthesis; it uses O2 to breakdown glucose into CO2 and H2O. It results in chemical cycling in biosphere. Respiration and Breathing: Respiration takes place in cells and needs O2 to breakdown food and releases the waste matter CO2. Breathing exchanges these gases between lungs and air. Overall equation for cellular respiration is: C6H1206 + O2 6CO2 + 6 H2O + ATP Glucose Oxygen Carbon Dioxide Water Energy Redox reactions: reduction-oxidation reactions. The gain of electrons during a chemical reaction is called Reduction. The loss of electrons during a chemical reaction is called Oxidation. Glucose is oxidized to 6CO2 and O2 is reduced to 6H2O during cellular respiration. During cellular respiration, glucose loses electrons and H, and O2 gains them. Energy and Food All living things need energy. Some living things can make their food from CO2 and H2O – Producers (plants, algae) Animals feeding on plants – herbivores (chipmunk) Animals feeding on animals – Carnivores (lion) Producers change solar energy to chemical energy of organic molecules – glucose, amino acids Animals and also plants break chemical bonds of sugar molecules and make ATP. Use ATP for all cellular functions 4 Main Step of Cellular Respiration Glycolysis: Glucose + 2NAD + 2ADP 2 Pyruvate + 2NADH + 2 ATP Preparatory Step: Pyruvate + NAD Acetyl-CoA + CO2 + NADH Krebs Cycle: Acetyl-CoA + NAD + FAD + ADP CO2 + NADH + FADH + ATP Electron Transport Chain: electrons of NADH + O2 ATP + H2O Cellular Respiration Aerobic Harvest of energy: is the main source of energy for most organisms.
    [Show full text]