Properties of succinyl-coenzyme A:L-malate coenzyme A transferase and its role in the autotrophic 3-hydroxypropionate cycle of Chloroflexus aurantiacus.
The 3-hydroxypropionate cycle has been proposed to operate as the autotrophic CO2 fixation pathway in the phototrophic bacterium Chloroflexus aurantiacus. In this pathway, acetyl coenzyme A (acetyl-CoA) and two bicarbonate molecules are converted to malate. Acetyl-CoA is regenerated from malyl-CoA by L-malyl-CoA lyase. The enzyme forming malyl-CoA, succinyl-CoA:L-malate coenzyme A transferase, was purified. Based on the N-terminal amino acid sequence of its two subunits, the corresponding genes were identified on a gene cluster which also contains the gene for L-malyl-CoA lyase, the subsequent enzyme in the pathway. Both enzymes were severalfold up-regulated under autotrophic conditions, which is in line with their proposed function in CO2 fixation. The two CoA transferase genes were cloned and heterologously expressed in Escherichia coli, and the recombinant enzyme was purified and studied. Succinyl-CoA:L-malate CoA transferase forms a large (alphabeta)n complex consisting of 46- and 44-kDa subunits and catalyzes the reversible reaction succinyl-CoA + L-malate --> succinate + L-malyl-CoA. It is specific for succinyl-CoA as the CoA donor but accepts L-citramalate instead of L-malate as the CoA acceptor; the corresponding d-stereoisomers are not accepted. The enzyme is a member of the class III of the CoA transferase family. The demonstration of the missing CoA transferase closes the last gap in the proposed 3-hydroxypropionate cycle.
Mikrobiologie, Institut Biologie II, Schänzlestr. 1, D-79104 Freiburg, Germany.
Silke
S
Friedmann
Astrid
A
Steindorf
Birgit E
BE
Alber
Georg
G
Fuchs
D000214:Acyl Coenzyme A / Q000378:metabolism*; D002384:Catalysis; D041861:Chloroflexus / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D015256:Coenzyme A-Transferases / Q000378:metabolism*; D019344:Lactic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D021122:Protein Subunits; D014166:Transferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D000214:Acyl Coenzyme A; D021122:Protein Subunits; D019344:Lactic Acid; C012046:succinyl-coenzyme A; C031601:hydracrylic acid; D014166:Transferases; D015256:Coenzyme A-Transferases; C509675:L-malate coenzyme A transferase