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Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
Published on: June 20, 2019
Acetyl-CoA decarbonylase/synthase complex from Archaeoglobus fulgidus
Y R Dai1, D W Reed, J H Millstein
1Department of Biochemistry and Molecular Biology, Uniformed Services University of Health Sciences, Bethesda, MD 20814-4799, USA.
The acetyl-CoA decarbonylase/synthase (ACDS) enzyme complex, typically found in methanogens, was discovered in Archaeoglobus fulgidus. This finding reveals a functional ACDS in a non-methanogenic organism, expanding our understanding of acetyl-CoA metabolism.
Area of Science:
- Biochemistry
- Microbiology
- Enzymology
Background:
- The acetyl-CoA decarbonylase/synthase (ACDS) multienzyme complex is crucial for acetyl-CoA metabolism in methanogenic archaea.
- Its presence and function in non-methanogenic organisms remain largely unexplored.
Purpose of the Study:
- To investigate the existence and function of the ACDS enzyme complex in the archaeon Archaeoglobus fulgidus.
- To characterize the subunits and catalytic activity of the ACDS complex from A. fulgidus.
Main Methods:
- Biochemical assays were performed to detect and analyze the ACDS activity in cell extracts of Archaeoglobus fulgidus.
- The enzyme complex was purified and its subunit composition was determined using SDS-PAGE.
Main Results:
- A functional ACDS multienzyme complex was identified in Archaeoglobus fulgidus, a non-methanogen.
- The A. fulgidus ACDS complex consists of five subunits with molecular masses of 89, 72, 50, 49.5, and 18.5 kDa.
- The complex catalyzed the synthesis of acetyl-CoA from CO2, reduced ferredoxin, N5-methyl-tetrahydrosarcinapterin, and CoA.
Conclusions:
- The study demonstrates the presence of a functional acetyl-CoA decarbonylase/synthase complex in a non-methanogenic archaeon.
- This finding broadens the known phylogenetic distribution of the ACDS complex and its role in archaeal metabolism.
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