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Bioorganic
Chemistry and Bio-Crystallography laboratory (B2Cl)
Main line of
research is the study of the relationship between the plant pathogen Erwinia amylovora and
the host Malus. spp.
E. amylovora is the bacterium causing fire
blight, a disease representing a threat to apple and pear farming worldwide
This is the link to the
Province website about fire blight, in Italian colpo di fuoco
The table below reports the
status of some of the targets
Target PCR Expression vector Protein expression Protein purification Crystals Structure solved HrpN √ √ HrpW √ √ √ √ DspB √ √ √ √ √ Lsc √ √ √ √ √ √ Avrpt2 √ √ √ √ √ √ GalE √ √ √
AmsI √ √ √ √ √ √ AmsB √ √ √ AmsD √ √ √ AmsE √ √ √ AmsK √ √ √ DfoA √ √ √ √ √ √ DfoC √ √ √ √ √ √ DfoJ √ √ √ √ √ √ GalU √ √ √ √ √ √ SrlD √ √ √ √ √ √ AmyR √ √ √ √ √ √
ViuB
√
√
√
√
√
FhuD
√
√
√
Projects funded:
·
EraSe = Erwinia amylovora’s Secretome (42800 €, 3 Years funding provided by the Free University of Bolzano from
01/11/2011)
E. amylovora’s secretome is the collection
of the proteins secreted by the pathogen. Many years of work on E. amylovora
demonstrated the importance of its secretome in host
colonization and disease development. Even though E. amylovora ‘s secretome has been shown to be
the key for a successful pathogenesis, yet very little is known about the
structure and mode of action of its component. Aim of this project is the study of proteins of E.amylovora’s secretome
involved in pathogenicity by means of gene cloning, protein expression,
purification and characterization and structure determination by X-ray
crystallography. From this project a collaboration with Dr
Malnoy's research group (FEM-IASMA) has been started.
· ViPEra = Virulence and Pathogenesis of Erwinia amylovora. A Structural genomics approach for the study of Erwinia amylovora virulence and pathogenesis. (179600 €, 3 Years funding provided by the Province of Bolzano from 01/11/2011 and with an additional contribution of a one year postdoctoral fellowship (25389€) from the FUB Foundation to study the enzymes DfoA, DfoC and DfoJ involved in desferrioxamine biosynthesis).
The proposed research deals with the study of proteins that are important in the virulence and pathogenesis of E. amylovora, using a “quasi”structural genomics approach. The “quasi” means that the proteins of interest will also be biochemically characterized in vitro by NMR and other analytical techniques.
The recent determination of the complete sequence of the genome of E. amylovora
enables to use the information contained in the bacterial DNA to select and
study the proteins crucial for disease thriving. Some pathogenesis factors have
been identified including the exopolysaccharide amylovoran and the Hrp proteins (hypersensitive reaction and pathogenicity).
The project’s aim
is to study the proteins involved in the biosynthesis of the exopolysaccharide amylovoran
(about 12 proteins), other enzymes involved in the methabolism
of sugars and the proteins involved in the biosynthesis of the siderophore desferrioxamine.
·
GAMEs:
Galactose and glucuronic Adic
Metabolism in Erwinia spp (31822 €, 2 Years funding provided by the Free University of Bolzano from
01/10/2013) and with an additional contribution of a one year postdoctoral
fellowship (25389 €) from the FUB Foundation to study GalU
and the enzymes involved in sorbitol metabolism.
The project’s aim is
to study the enzymes UDP-galactose 4-epimerase (GalE;
EC 5.1.3.2), that synthetize UDP-galactose from UDP-glucose and UDP-glucose
dehydrogenase (Ugd; EC 1.1.1.22), that converts
UDP-glucose into UDP-glucuronic acid.
GalE and Ugd
are key enzymes essential in the biosynthesis of amylovoran.
In E. amylovora
GalE is constitutively expressed and its importance
was demonstrated when a GalE- mutant was shown to be deficient in amylovoran synthesis and showed loss of virulence on pear
fruits and seedlings
·
Mescal: Molecular Engineering of
the Structural and Catalytic properties of Levansucrase
(72000€, 3 Years funding
provided by the Free University of Bolzano from 01/01/2015)
The
research project is divided in three workpackages:
1)
Structural and functional characterization of the levansucrase
from the non-pathogenic epiphytic Erwinia tasmaniensis, comparison with the levansucrase
from the phytopathogenic Erwinia amylovora and characterization of the
carbohydrates they synthesise.
2)
Molecular engineering of levansucrase. Structural and
functional characterization of the engineered enzyme. Characterization of the
carbohydrates produced by the engineered levansucrases.
3)
Bioinformatics, phylogenetic analysis and mathematical modelling of fructose
polymerization.
UREASE
A long standing project in collaboration with Professor Ciurli is the study of the structure and function relationship of the enzyme urease from Sporosarcina pasteurii (previously known as Bacillus pasteurii).