SBS-713

Archaea, the third domain of life

Metabolism and Metabolic enzymes in

Archaea
Glycolysis and Gluconeogenesis

Dr. Mehwish Aslam

Last lecture
• EM Pathway
6PGD (EDD), gluconate-6-phosphate dehydratase
ENO, enolase;
G6PDH (Zwf), glucose-6-phosphate dehydrogenase;
GAD, gluconate dehydratase; Nonphosphorylative ED
GADH, glyceraldehyde dehydrogenase; (npED)
GAOR, glyceraldehyde:ferredoxin oxidoreductase;
GAPDH, glyceraldehyde-3-phosphate dehydrogenase;
GAPN, nonphosphorylating GAPDH;
GAPOR, GAP:Fd oxidoreductase;
GDH, glucose dehydrogenase;
GK, glycerate kinase;
GLac, gluconolactonase;
GLK, glucokinase;
KDGK, 2-keto-3-deoxygluconate kinase;
KDPGA (EDA), 2-keto-3-deoxy-6-phosphogluconate aldolase; Semiphosphorylative ED
KD(P)GA, 2-keto-3-deoxy-(6-phospho)gluconate aldolase; (spED)
PGK, phosphoglycerate kinase;
PGL, 6-phosphoglucono-1,4-lactonase;
PGAM, phosphoglycerate mutase
PK, pyruvate kinase;
PEPS, PEP synthetase;
PPDK, pyruvate:phosphate dikinase;
PTS, PEP-dependent phosphotransferase system;
KDG, 2-keto-3-deoxygluconate;
KDPG, 2-keto-3-deoxy-6-phosphogluconate
Glucose Phosphorylation

• Glucose à glucose 6-phosphate

• In Bacteria
• Glucose transport occurs via the PEP Phosphotransferase
• catalyzed by glucose and ATP-specific kinases
• In Eukarya
• catalyzed by ATP-specific hexokinases
• Both the eukaryotic hexokinases as well as the bacterial glucokinases
constitute distinct families within the actin-like ATPase domain superfamily.

• In Archaea, two different mechanisms of glucose phosphorylation have been

described.
ADP- and ATP-dependent glucokinase.
Euryarchaeota

• Pyrococcus furiosus, Thermococcus litoralis, and Archaeoglobus fulgidus,

Thermococcus chitonophagus, glucose phosphorylation is carried out by ADP-
dependent glucokinases (ADP- GLKs).
• These enzymes show high specificity for glucose and belong to the ribokinase
superfamily.
• Glucokinase (GK) enzyme of Pyrococcus furiosus and Thermococcus chitonophagus is a
Bifunctional enzyme that possesses additional glucosamine kinase (Gln kinase)
activity.
• Archaeal ADP-GLKs prefer ADP as the phosphoryl donor and additionally also utilize
CDP but not GDP, IDP, or UDP.
• The Bifunctional ADP-GLK/PFK from Methanocaldococcus jannaschii also utilizes GDP
(40%) in addition to ADP (100%) and, less efficiently, CDP (14%)
ADP- and ATP-dependent glucokinase
Crenarchaeota

• The second mechanism of glucose phosphorylation in Archaea is catalyzed by ATP-

dependent kinases with broad sugar substrate specificity.
• In Thermoproteus tenax, this process is ATP dependent (ADP forming), involving an
ATP-dependent hexokinase with a broad hexose substrate spectrum
• These ATP-dependent archaeal hexokinases belong to the ROK (repressor protein,
open reading frame, sugar kinase) family.
• Eukaryotic hexokinases à Monomeric, and sometimes dimeric (subunit of ~50-100
kDa)
• Bacterial ATP-dependent glucokinases à dimers composed of ~30-kDa subunits,
• Archaeal ADP-GLK have been biochemically characterized as monomeric enzymes
(subunit of ~50 kDa)
ADP-forming acetyl-CoA synthetase (ADP-forming) (ACD) acetyl-CoA + ADP + Pi = acetate + ATP + CoA