The LPG1x family from Leishmania major is constituted of rare eukaryotic galactofuranosyltransferases with unprecedented catalytic properties

Abstract

Galactofuranosyltransferases are poorly described enzymes despite their crucial role in the virulence and the pathogenicity of numerous microorganisms. These enzymes are considered as potential targets for therapeutic action. In addition to the only well-characterised prokaryotic GlfT2 from Mycobacterium tuberculosis, four putative genes in Leishmania major were previously described as potential galactofuranosyltransferases. In this study, we have cloned, over-expressed, purified and fully determined the kinetic parameters of these four eukaryotic enzymes, thus demonstrating their unique potency in catalysing the transfer of the galactofuranosyl moiety into acceptors. Their individual promiscuity revealed to be different, as some of them could efficiently use NDP-pyranoses as donor substrates in addition to the natural UDP-galactofuranose. Such results pave the way for the development of chemoenzymatic synthesis of furanosyl-containing glycoconjugates as well as the design of improved drugs against leishmaniasis.

Galactofuranosidase from JHA 19 Streptomyces sp.: subcloning and biochemical characterization

“…[1] However, under its furanose form, galactofuranose (Galf) is totally absent from mammals, but is present in many pathogenic microorganisms. [2] , [3] Indeed, there are three main and specific enzymes involved in the biosynthesis and the metabolism of the Galfcontaining molecules [4]: UDP-galactopyranose mutase (UGM), [5] galactofuranosyltransferase (GalfT) [6] and galactofuranosidase (Galf-ase). [7] The glycobiology of Galf and enzymes involved in its metabolism have thus become attractive targets for the development of antimicrobial agents.…”

Galactofuranose-Related Enzymes: Challenges and Hopes

“…Until recently, LPG1 from Leishmania major was the first and only described eukaryotic Galf T. In an extensive study from 2018, all the four putative transferases (LPG1, LPG1G, LPG1L and LPG1R) encoded in the L. major genome were cloned, overexpressed and their kinetic parameters determined. It was demonstrated that they are able to use both UDP-Galf and UDP-Galp as donor substrates [42].…”