Biologia

, 66:954 | Cite as

Comparison of dynamic responses of cellular metabolites in Escherichia coli to pulse addition of substrates

  • Md. Aminul Hoque
  • Atefeh Taherian Fard
  • Mosfequr Rahman
  • Omar Alattas
  • Kohei Akazawa
  • Amir Feisal Merican
Section Cellular and Molecular Biology
First Online:
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Abstract

We conducted an integrated study of cell growth parameters, product formation, and the dynamics of intracellular metabolite concentrations using Escherichia coli with genes knocked out in the glycolytic and oxidative pentose phosphate pathway (PPP) for glucose catabolism. We investigated the same characteristics in the wild-type strain, using acetate or pyruvate as the sole carbon source. Dramatic effects on growth parameters and extracellular and intracellular metabolite concentrations were observed after blocking either glycolytic breakdown of glucose by inactivation of phosphoglucose isomerase (disruption of pgi gene) or pentose phosphate breakdown of glucose by inactivation of glucose-6-phosphate dehydrogenase (disruption of zwf gene). Reducing power (NADPH) was mainly produced through PPP when the pgi gene was knocked out, while NADPH was produced through the tricarboxylic acid (TCA) cycle by isocitrate dehydrogenase or NADP-linked malic enzyme when the zwf gene was knocked out. As expected, when the pgi gene was knocked out, intracellular concentrations of PPP metabolites were high and glycolytic and concentrations of TCA cycle pathway metabolites were low. In the zwf gene knockout, concentrations of PPP metabolites were low and concentrations of intracellular glycolytic and TCA cycle metabolites were high.

Key words

pulse addition dynamic response metabolic engineering gene knockout pgi mutant zwf mutant metabolite concentration 

Abbreviations

CER

CO2 evolution rate

DCW

dry cell weight

ED

Entner-Doudoroff

EMP

Embden-Mayerhof-Paranas

MEZ

malic enzyme

OD

optical density

OUR

oxygen uptake rate

PGI

phosphoglucose isomerase

PPP

pentose phosphate pathway

TCA

tricarboxylic acid

Yx/s

cell mass yield

Nomenclature: Enzymes

6PGDH

6-phosphogluconate dehydrogenase

Eda

Entner-Douderoff aldolase

Edd

Entner-Douderoff dehydralase

Eno

enolase

Fba

fructose-1,6-bisphosphate aldolase

G6PDH

glucose-6-phosphate dehydrogenase

GAPDH

glyceraldehyde-3-phosphate dehydrogenase

GDH

glutamate dehydrogenase

Hxk

hexokinase

ICDH

isocitrate dehydrogenase

LDH

lactate dehydrogenase

MDH

malate dehydrogenase

Mk

myokinase

Pck

PEP carboxykinase

Pgi

phosphoglucose isomerase

Ppc

phosphoenolpyruvate carboxylase

Pta

phosphotransacetylase

PTS

phosphotransferase system

Pyk

pyruvate kinase

Rpe

ribose-phosphate epimerase

Rpi

ribose-phosphate isomerase

Tkt

transketolase

Tpi

triosephosphate isomerase

Tal

transaldolase

Metabolites

2PG

2-phosphoglycerate

6PG

6-phosphogluconate

AcCoA

acetyl-coenzyme A

ADP

adenosine diphosphate

AKG

α-ketoglutarate

AMP

adenosine monophosphate

ATP

adenosine triphosphate

DHAP

dihydroxyacetone phosphate

E4P

erythrose-4-phosphate

F6P

fructose-6-phosphate

FBP

fructose-1,6-bisphosphate

G6P

glucose-6-phosphate

GAP

glyceraldehyde-3-phosphate

ICT

isocitrate

NAD

diphosphopyridindinucleotide, oxidized

NADH

diphosphopyridindinucleotide, reduced

NADP

diphosphopyridindinucleotide-phosphate, oxidized

NADPH

diphosphopyridindinucleotide-phosphate, reduced

OAA

oxaloacetate

PEP

phosphoenolpyruvate

PYR

pyruvate

R5P

ribose-5-phosphate

RU5P

ribulose-5-phosphate

SUC

succinate

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Copyright information

© © Versita Warsaw and Springer-Verlag Wien 2011

Authors and Affiliations

  • Md. Aminul Hoque
    • 1
    • 2
    • 3
  • Atefeh Taherian Fard
    • 1
  • Mosfequr Rahman
    • 3
  • Omar Alattas
    • 4
  • Kohei Akazawa
    • 2
  • Amir Feisal Merican
    • 1
  1. 1.Centre of Research in Computational Sciences and Informatics in Biology, Biodiversity, Environment, Agriculture and Healthcare (CRYSTAL), Institute of Biological Sciences, Faculty of ScienceUniversity of MalayaKuala LumpurMalaysia
  2. 2.Division of Information Science and Biostatistics, Department of Medical InformaticsNiigata University Medical and Dental HospitalNiigataJapan
  3. 3.Faculty of ScienceRajshahi UniversityRajshahiBangladesh
  4. 4.Centre of Excellence in Biotechnology ResearchKing Saud UniversityRiyadhKingdom of Saudi Arabia

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