Recombinant Escherichia coli O9:H4 Cardiolipin synthase (cls)
Description
Definition of Recombinant Escherichia coli O9:H4 Cardiolipin Synthase (Cls)
Recombinant Escherichia coli O9:H4 Cardiolipin synthase (Cls) is an enzyme that catalyzes the synthesis of cardiolipin in Escherichia coli . Cardiolipin is a unique dimeric phospholipid primarily located in the inner mitochondrial membrane of eukaryotes and the bacterial plasma membrane of prokaryotes . In E. coli, cardiolipin synthase (Cls) facilitates the transfer of phosphatidyl groups between phosphatidylglycerol molecules, resulting in cardiolipin and glycerol . The enzyme is encoded by the cls gene, situated at minute 28.02 on the E. coli genetic map .
Function and Mechanism
E. coli cardiolipin synthase converts two phosphatidylglycerol molecules into cardiolipin and glycerol . The enzyme uses a mixed micelle assay in which phosphatidyl[2-3H]glycerol acts as the substrate . Cardiolipin synthase activity is stimulated by phosphate and Triton X-100 .
Production and Purification
Recombinant DNA technology allows for the amplification and purification of cardiolipin synthase . For example, the cls gene can be inserted downstream from a T7 RNA promoter in a recombinant plasmid, such as pLR3 . When this plasmid is introduced into E. coli strain BL21(DE3), the resulting strain, BL21(DE3)/pLR3, exhibits significantly elevated cardiolipin synthase activity compared to wild-type cells . The enzyme can be purified to homogeneity through extraction with Triton X-114 and chromatography on DEAE-cellulose .
Biological Significance
Mutations in the cls gene can lead to several phenotypic changes in E. coli, including longer doubling times, decreased viability during the stationary phase, increased resistance to 3,4-dihydroxybutyl-1-phosphonate, and altered sensitivity to novobiocin . Cardiolipin synthase binds to anionic phospholipids, especially cardiolipin, which inhibits its phosphotransfer activity . Cardiolipin inhibition of cardiolipin synthase likely plays an important role in regulating cardiolipin synthesis in E. coli .
Applications
Recombinant E. coli O9:H4 Cardiolipin synthase (Cls) is available for purchase for research purposes . It is produced in an in vitro E. coli expression system with high purity and is commonly used in studies related to lipid metabolism and enzyme regulation .
Product Specs
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Target Background
Function: Catalyzes the reversible transfer of phosphatidyl groups between phosphatidylglycerol molecules, resulting in the formation of cardiolipin (CL, diphosphatidylglycerol) and glycerol.
KEGG: ecx:EcHS_A1358
