Systematic identification and elimination of flux bottlenecks in the aldehyde production pathway of Synechococcus elongatus PCC 7942.

Acetolactate Synthase / metabolism Aldehydes / metabolism Amino Acids / metabolism Metabolic Engineering Metabolic Flux Analysis Phosphoenolpyruvate Carboxylase / metabolism Pyruvate Dehydrogenase Complex / metabolism Pyruvate Kinase / metabolism Pyruvates / metabolism RNA, Bacterial / biosynthesis Synechococcus / metabolism
Cyanobacteria Isobutyraldehyde Metabolic flux analysis Phosphoenolpyruvate Photoautotrophic metabolism Pyruvate Stable isotope

Journal

Metabolic engineering
ISSN: 1096-7184
Titre abrégé: Metab Eng
Pays: Belgium
ID NLM: 9815657

Informations de publication

Date de publication:
07 2020
Historique:
received: 03 12 2019
revised: 27 02 2020
accepted: 17 03 2020
pubmed: 31 3 2020
medline: 29 5 2021
entrez: 31 3 2020
Statut: ppublish

Résumé

Isotopically nonstationary metabolic flux analysis (INST-MFA) provides a versatile platform to quantitatively assess in vivo metabolic activities of autotrophic systems. By applying INST-MFA to recombinant aldehyde-producing cyanobacteria, we identified metabolic alterations that correlated with increased strain performance in order to guide rational metabolic engineering. We identified four reactions adjacent to the pyruvate node that varied significantly with increasing aldehyde production: pyruvate kinase (PK) and acetolactate synthase (ALS) fluxes were directly correlated with product formation, while pyruvate dehydrogenase (PDH) and phosphoenolpyruvate carboxylase (PPC) fluxes were inversely correlated. Overexpression of enzymes for PK or ALS did not result in further improvements to the previous best-performing strain, while downregulation of PDH expression (through antisense RNA expression) or PPC flux (through expression of the reverse reaction, phosphoenolpyruvate carboxykinase) provided significant improvements. These results illustrate the potential of INST-MFA to enable a systematic approach for iterative identification and removal of pathway bottlenecks in autotrophic host cells.

Identifiants

DOI: 10.1016/j.ymben.2020.03.007 PMID: 32222320 PMC: PMC7217728
pubmed: 32222320
pii: S1096-7176(20)30062-8
doi: 10.1016/j.ymben.2020.03.007
pmc: PMC7217728
mid: NIHMS1583844
pii:
doi:

Substances chimiques

Aldehydes 0
Amino Acids 0
Pyruvate Dehydrogenase Complex 0
Pyruvates 0
RNA, Bacterial 0
Acetolactate Synthase EC 2.2.1.6
Pyruvate Kinase EC 2.7.1.40
Phosphoenolpyruvate Carboxylase EC 4.1.1.31

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Sous-ensembles de citation

IM

Pagination

56-65

Subventions

Organisme : NIGMS NIH HHS
ID : R01 GM067152
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM107434
Pays : United States
Organisme : NIGMS NIH HHS
ID : R37 GM067152
Pays : United States
Organisme : NIDDK NIH HHS
ID : T32 DK101003
Pays : United States

Informations de copyright

Copyright © 2020 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Références

Proc Natl Acad Sci U S A. 2014 Nov 25;111(47):16967-72
pubmed: 25368168
Curr Opin Biotechnol. 2013 Dec;24(6):1116-21
pubmed: 23453397
Anal Chem. 2014 Feb 18;86(4):2221-8
pubmed: 24498896
Photosynth Res. 2015 Oct;126(1):19-32
pubmed: 25280933
Cell Mol Life Sci. 2007 Apr;64(7-8):892-905
pubmed: 17429582
Nucleic Acids Res. 1995 Jul 11;23(13):2472-8
pubmed: 7630724
Biotechnol Biofuels. 2017 Nov 16;10:273
pubmed: 29177008
Appl Environ Microbiol. 2013 Sep;79(18):5566-75
pubmed: 23835179
Biomed Res Int. 2014;2014:627014
pubmed: 25006579
Appl Environ Microbiol. 2013 Feb;79(4):1250-7
pubmed: 23241971
Metabolism. 2014 Feb;63(2):283-95
pubmed: 24286856
Trends Biotechnol. 2015 Jun;33(6):352-61
pubmed: 25908503
Biotechnol Bioeng. 2013 Jun;110(6):1748-58
pubmed: 23296385
ACS Synth Biol. 2015 Nov 20;4(11):1197-204
pubmed: 25974153
J Mass Spectrom. 1996 Mar;31(3):255-62
pubmed: 8799277
Curr Opin Biotechnol. 2015 Dec;36:50-6
pubmed: 26319894
Methods Mol Biol. 2014;1090:181-210
pubmed: 24222417
PLoS One. 2011 Jan 21;6(1):e16278
pubmed: 21283704
Nucleic Acids Res. 2007 Jul;35(Web Server issue):W71-4
pubmed: 17485472
Phytochemistry. 2007 Aug-Sep;68(16-18):2302-12
pubmed: 17524438
Metab Eng. 2019 Jul;54:222-231
pubmed: 31029860
Plant Physiol. 2015 Feb;167(2):586-99
pubmed: 25511434
J Biotechnol. 2014 Aug 20;184:100-2
pubmed: 24858679
Microb Cell Fact. 2016 Nov 15;15(1):196
pubmed: 27846887
Metab Eng. 2006 Jul;8(4):324-37
pubmed: 16631402
EMBO J. 2000 Jul 3;19(13):3349-57
pubmed: 10880447
Appl Microbiol Biotechnol. 2018 Jul;102(13):5457-5471
pubmed: 29744631
J Biol Chem. 1994 Feb 18;269(7):5122-6
pubmed: 8106492
Nat Biotechnol. 2009 Dec;27(12):1177-80
pubmed: 19915552
J Bacteriol. 2008 Jun;190(11):3851-8
pubmed: 18375566
Cell. 2016 Mar 10;164(6):1185-1197
pubmed: 26967285
Bioinformatics. 2014 May 1;30(9):1333-5
pubmed: 24413674
Biotechnol Bioeng. 2008 Feb 15;99(3):686-99
pubmed: 17787013
PLoS Comput Biol. 2010 Apr 15;6(4):e1000744
pubmed: 20419153
Appl Environ Microbiol. 2010 Apr;76(7):2107-14
pubmed: 20118372
Nature. 2013 Mar 7;495(7439):116-20
pubmed: 23417065
ACS Synth Biol. 2018 Jul 20;7(7):1669-1675
pubmed: 29874914
Anal Chem. 2007 Oct 1;79(19):7554-9
pubmed: 17822305
Metab Eng. 2011 Nov;13(6):656-65
pubmed: 21907300
Metab Eng. 2017 Jul;42:9-18
pubmed: 28479191
J Ind Microbiol. 1996 Aug;17(2):80-3
pubmed: 8987894
Proc Natl Acad Sci U S A. 2016 Dec 20;113(51):E8344-E8353
pubmed: 27911809
Biotechnol Bioeng. 2006 Jun 5;94(2):234-51
pubmed: 16598793
Proc Natl Acad Sci U S A. 2012 Feb 14;109(7):2678-83
pubmed: 22308420
Biotechnol Prog. 2005 Nov-Dec;21(6):1617-26
pubmed: 16321043
Methods Mol Biol. 2014;1083:85-108
pubmed: 24218212
Appl Environ Microbiol. 1999 Feb;65(2):523-8
pubmed: 9925577
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
Plant Physiol. 1993 Jan;101(1):121-6
pubmed: 8278492
Adv Microb Physiol. 1989;30:183-238
pubmed: 2700539
Curr Opin Biotechnol. 2018 Dec;54:80-87
pubmed: 29522915
Anal Chem. 2018 Jun 5;90(11):6486-6493
pubmed: 29712418
FEMS Microbiol Lett. 2004 Sep 15;238(2):367-74
pubmed: 15358422
Biotechnol Bioeng. 2003 Dec 20;84(6):647-57
pubmed: 14595777
J Bacteriol. 2005 Sep;187(17):6005-18
pubmed: 16109942
Biotechnol Bioeng. 2015 Oct;112(10):2195-9
pubmed: 25899755

Auteurs

Yi Ern Cheah (YE)

Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, USA.

Yao Xu (Y)

Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA.

Sarah A Sacco (SA)

Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, USA.

Piyoosh K Babele (PK)

Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, USA.

Amy O Zheng (AO)

Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, USA.

Carl Hirschie Johnson (CH)

Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, USA.

Jamey D Young (JD)

Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, USA; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, USA. Electronic address: j.d.young@vanderbilt.edu.

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Flavoprotéines › Acetolactate synthase › Systematic identification and elimination of...
questionsmedicales.fr › Composés chimiques organiques › Aldéhydes › Systematic identification and elimination of...
questionsmedicales.fr › Acides aminés, peptides et protéines › Acides aminés › Systematic identification and elimination of...
questionsmedicales.fr › Technologie, industrie et agriculture › Ingénierie › Bioingénierie › Ingénierie cellulaire › Génie métabolique › Systematic identification and elimination of...
questionsmedicales.fr › Techniques d'investigation › Techniques cytologiques › Analyse des flux métaboliques › Systematic identification and elimination of...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Lyases › Carbon-carbon lyases › Carboxy-lyases › Phosphoenolpyruvate carboxylase › Systematic identification and elimination of...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Complexes multienzymatiques › Complexe du pyruvate déshydrogénase › Systematic identification and elimination of...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Transferases › Phosphotransferases › Phosphotransferases (Alcohol Group Acceptor) › Pyruvate kinase › Systematic identification and elimination of...
questionsmedicales.fr › Composés chimiques organiques › Acides carboxyliques › Cétoacides › Pyruvates › Systematic identification and elimination of...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN bactérien › Systematic identification and elimination of...
questionsmedicales.fr › Bactéries › Bactéries à Gram négatif › Bactéries à Gram négatif photosynthétiques › Cyanobactéries › Synechococcus › Systematic identification and elimination of...