Inorganic pyrophosphatase in uncultivable hemotrophic mycoplasmas: identification and properties of the enzyme from Mycoplasma suisReportar como inadecuado




Inorganic pyrophosphatase in uncultivable hemotrophic mycoplasmas: identification and properties of the enzyme from Mycoplasma suis - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

BMC Microbiology

, 10:194

First Online: 20 July 2010Received: 27 October 2009Accepted: 20 July 2010

Abstract

BackgroundMycoplasma suis belongs to a group of highly specialized hemotrophic bacteria that attach to the surface of host erythrocytes. Hemotrophic mycoplasmas are uncultivable and the genomes are not sequenced so far. Therefore, there is a need for the clarification of essential metabolic pathways which could be crucial barriers for the establishment of an in vitro cultivation system for these veterinary significant bacteria.

Inorganic pyrophosphatases PPase are important enzymes that catalyze the hydrolysis of inorganic pyrophosphate PPi to inorganic phosphate Pi. PPases are essential and ubiquitous metal-dependent enzymes providing a thermodynamic pull for many biosynthetic reactions. Here, we describe the identification, recombinant production and characterization of the soluble sPPase of Mycoplasma suis.

ResultsScreening of genomic M. suis libraries was used to identify a gene encoding the M. suis inorganic pyrophosphatase sPPase. The M. suis sPPase consists of 164 amino acids with a molecular mass of 20 kDa. The highest identity of 63.7% was found to the M. penetrans sPPase. The typical 13 active site residues as well as the cation binding signature could be also identified in the M. suis sPPase. The activity of the M. suis enzyme was strongly dependent on Mg and significantly lower in the presence of Mn and Zn. Addition of Ca and EDTA inhibited the M. suis sPPase activity. These characteristics confirmed the affiliation of the M. suis PPase to family I soluble PPases. The highest activity was determined at pH 9.0. In M. suis the sPPase builds tetramers of 80 kDa which were detected by convalescent sera from experimentally M. suis infected pigs.

ConclusionThe identification and characterization of the sPPase of M. suis is an additional step towards the clarification of the metabolism of hemotrophic mycoplasmas and, thus, important for the establishment of an in vitro cultivation system. As an antigenic and conserved protein the M. suis sPPase could in future be further analyzed as a diagnostic antigen.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2180-10-194 contains supplementary material, which is available to authorized users.

Download fulltext PDF



Autor: Katharina Hoelzle - Simone Peter - Michele Sidler - Manuela M Kramer - Max M Wittenbrink - Kathrin M Felder - Ludwig E 

Fuente: https://link.springer.com/article/10.1186/1471-2180-10-194







Documentos relacionados