title: Recombinant broad-range phospholipase C from Listeria monocytogenes exhibits optimal activity at acidic pH.
abstract: The broad-range phospholipase C (PLC) from Listeria monocytogenes has been expressed using an intein expression system and characterized. This zinc metalloenzyme, similar to the homologous enzyme from Bacillus cereus, targets a wide range of lipid substrates. With monomeric substrates, the length of the hydrophobic acyl chain has significant impact on enzyme efficiency by affecting substrate affinity (Km). Based on a homology model of the enzyme to the B. cereus protein, several active site residue mutations were generated. While this PLC shares many of the mechanistic characteristics of the B. cereus PLC, a major difference is that the L. monocytogenes enzyme displays an acidic pH optimum regardless of substrate status (monomer, micelle, or vesicle). This unusual behavior might be advantageous for its role in the pathogenicity of L. monocytogenes.
authors.affiliation: Department of Chemistry, Boston College, Chestnut Hill, MA, United States. Electronic address: huangqa@bc.edu.
authors.forename: Qiongying
authors.initials: Q
authors.lastname: Huang
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, MA, United States.
authors.forename: Anne
authors.initials: A
authors.lastname: Gershenson
authors.affiliation: Department of Chemistry, Boston College, Chestnut Hill, MA, United States.
authors.forename: Mary F
authors.initials: MF
authors.lastname: Roberts
mesh_terms: D000143:Acids / Q000378:metabolism*; D020134:Catalytic Domain; D006863:Hydrogen-Ion Concentration; D008089:Listeria monocytogenes / Q000201:enzymology*; D011994:Recombinant Proteins / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: D000143:Acids; D011994:Recombinant Proteins; D010738:Type C Phospholipases
keywords: Acidic pH optimum; Listeria monocytogenes; Mutagenesis; Phospholipase C; Vesicle binding