Characterization of acetyl-CoA/propionyl-CoA carboxylase in Metallosphaera sedula. Carboxylating enzyme in the 3-hydroxypropionate cycle for autotrophic carbon fixation.
-
The purified M. sedula carboxylase contains an 18.6 kDa biotin carrier protein (beta) together with the alpha and gamma subunits, probably as (alpha beta gamma)4.
"These subunits probably form an (alpha beta gamma)4 holoenzyme."
Presence of acetyl coenzyme A (CoA) carboxylase and propionyl-CoA carboxylase in autotrophic Crenarchaeota and indication for operation of a 3-hydroxypropionate cycle in autotrophic carbon fixation.
-
Autotrophic Sulfolobales, including M. sedula, contain large amounts of a small biotinylated protein. Archaea that use other autotrophic pathways lack it.
"Other archaea reported to use one of the other known autotrophic pathways lacked such small biotin-containing proteins."
Characterization of a bifunctional archaeal acyl coenzyme A carboxylase.
-
In the Acidianus brierleyi ortholog, the conserved biotin-binding site is on the 20 kDa BCCP subunit.
"the conserved biotin-binding site was located on the biotin carboxyl carrier protein"
Ancillary contributions of heterologous biotin protein ligase and carbonic anhydrase for CO<sub>2</sub> incorporation into 3-hydroxypropionate by metabolically engineered Pyrococcus furiosus.
-
The M. sedula biotin protein ligase Msed_2010 biotinylates the ACC beta subunit in vitro. Co-expressing it improves ACC activity and 3HP production in engineered P. furiosus.
"M. sedula BPL (Msed_2010) was shown to biotinylate the β-subunit (biotin carboxyl carrier protein) of the ACC in vitro."
-
Sulfolobales BCCPs, including M. sedula ACC-beta, have serine instead of methionine after the biotinylated lysine, which hinders biotinylation by E. coli and P. furiosus ligases.
"While similar biotinylated proteins in both E. coli and P. furiosus have a methionine immediately C-terminal to the biotinylated lysine residue, S. tokodaii, M. sedula and the other Sulfolobales have a serine"
A 3-hydroxypropionate/4-hydroxybutyrate autotrophic carbon dioxide assimilation pathway in Archaea.
Exploiting microbial hyperthermophilicity to produce an industrial chemical, using hydrogen and carbon dioxide.
-
Msed_0148 is E1-beta and is translationally coupled to Msed_0147 (E1-alpha).
"The M. sedula RBS for E1β was retained, as the two genes (E1α and E1β) appear to be translationally coupled."
Conversion of 4-hydroxybutyrate to acetyl coenzyme A and its anapleurosis in the Metallosphaera sedula 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway.
-
Msed_0148 is induced with the other ACC subunit genes under autotrophic growth.
"especially the genes encoding acetyl-CoA/propionyl-CoA carboxylase (ACCαβγ) (Msed_0147, Msed_0148, Msed_1375; 7- to 30-fold)"
UniProt record A4YD23 (A4YD23_METS5)