Propionyl coenzyme A (propionyl-CoA) carboxylase in Haloferax mediterranei: Indispensability for propionyl-CoA assimilation and impacts on global metabolism.
Propionyl coenzyme A (propionyl-CoA) is an important intermediate during the biosynthesis and catabolism of intracellular carbon storage of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in haloarchaea. However, the haloarchaeal propionyl-CoA carboxylase (PCC) and its physiological significance remain unclear. In this study, we identified a PCC that catalyzed propionyl-CoA carboxylation with an acetyl-CoA carboxylation side activity in Haloferax mediterranei. Gene knockout/complementation demonstrated that the PCC enzyme consisted of a fusion protein of a biotin carboxylase and a biotin-carboxyl carrier protein (PccA [HFX_2490]), a carboxyltransferase component (PccB [HFX_2478]), and an essential small subunit (PccX [HFX_2479]). Knockout of pccBX led to an inability to utilize propionate and a higher intracellular propionyl-CoA level, indicating that the PCC enzyme is indispensable for propionyl-CoA utilization. Interestingly, H. mediterranei DBX (pccBX-deleted strain) displayed multiple phenotypic changes, including retarded cell growth, decreased glucose consumption, impaired PHBV biosynthesis, and wrinkled cells. A propionyl-CoA concentration equivalent to the concentration that accumulated in DBX cells was demonstrated to inhibit succinyl-CoA synthetase of the tricarboxylic acid cycle in vitro. Genome-wide microarray analysis showed that many genes for glycolysis, pyruvate oxidation, PHBV accumulation, electron transport, and stress responses were affected in DBX. This study not only identified the haloarchaeal PCC for the metabolism of propionyl-CoA, an important intermediate in haloarchaea, but also demonstrated that impaired propionyl-CoA metabolism affected global metabolism in H. mediterranei.
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China University of Chinese Academy of Sciences, Beijing, China.
Jing
J
Hou
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China xiangh@im.ac.cn hanjing@im.ac.cn.
Hua
H
Xiang
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China xiangh@im.ac.cn hanjing@im.ac.cn.
Jing
J
Han
D000214:Acyl Coenzyme A / Q000378:metabolism*; D002244:Carbon / Q000378:metabolism; D020869:Gene Expression Profiling; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D005947:Glucose / Q000378:metabolism; D019623:Haloferax mediterranei / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000235:genetics; D043586:Methylmalonyl-CoA Decarboxylase / Q000235:genetics / Q000378:metabolism*; D011091:Polyesters / Q000378:metabolism*; D021122:Protein Subunits / Q000235:genetics; D000094122:Polyhydroxybutyrates
D000214:Acyl Coenzyme A; D002244:Carbon; D005947:Glucose; D043586:Methylmalonyl-CoA Decarboxylase; D011091:Polyesters; D021122:Protein Subunits; C052620:poly(3-hydroxybutyrate)-co-(3-hydroxyvalerate); D000094122:Polyhydroxybutyrates; C009061:propionyl-coenzyme A