Role of C-Terminal Domains in Surface Attachment of the Fructosyltransferase of Streptococcus salivariusATCC 25975

Rathsam, Catherine; Jacques, Nicholas A.

doi:10.1128/.180.23.6400-6403.1998

Cited by 5 publications

(5 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The purified Ftf gave an apparent M r of 125 400 on SDS\PAGE, 180 600 by gel filtration and 102 000 on native-PAGE (results not shown). N-terminal sequencing revealed that the Ftf expressed in E. coli, like that expressed by S. sali arius, had been cleaved at TQVKA DQVTET to remove its signal sequence [32]. The predicted M r of the Ftf devoid of its signal sequence is 98 450, which was close to the result obtained from the native-PAGE analysis.…”

Section: Purification and Characterization Of The Ftfsupporting

confidence: 69%

Purification and enzymic properties of the fructosyltransferase of Streptococcus salivarius ATCC 25975

Song

Jacques

1999

Biochem. J.

Self Cite

View full text Add to dashboard Cite

The recombinant fructosyltransferase (Ftf) of Streptococcus salivarius was expressed in Escherichia coli and purified to electrophoretic homogeneity after a combination of adsorption, ion-exchange and gel-filtration chromatography. The N-terminal signal sequence of the Ftf was removed by E. coli at the same site as in its natural host. The purified Ftf exhibited maximum activity at pH 6.0 and 37 degrees C, was activated by Ca2+, but inhibited by the metal ions Cu2+, Zn2+, Hg2+ and Fe3+. The enzyme catalysed the transfer of the fructosyl moiety of sucrose to a number of acceptors, including water, glucose and sucrose via a Ping Pong mechanism involving a fructosyl-enzyme intermediate. While this mechanism of catalysis is utilized by the levansucrases of Bacillus subtilis and Acetobacter diazotrophicus and the values of the kinetic constants for the three enzymes are similar, sucrose was a far more efficient fructosyl-acceptor for the Ftf of S. salivarius than for the two other enzymes.

show abstract

Section: Purification and Characterization Of The Ftfsupporting

confidence: 69%

Purification and enzymic properties of the fructosyltransferase of Streptococcus salivarius ATCC 25975

Song

Jacques

1999

Biochem. J.

Self Cite

View full text Add to dashboard Cite

show abstract

“…The cell associated FTF from Streptococcus salivarius which is devoid of motif LPXTG is released from the cells on exposure to sucrose. Through deletions within the C terminus of this enzyme, Rathsam and Jacques [ 18 ], implicated both the hydrophobic and the PGST-rich wall-associated domains in stabilizing the enzyme on the cell surface. In IslA, neither the LPXTG motif, nor the PGST motif is present.…”

Section: Resultsmentioning

confidence: 99%

Functional role of the additional domains in inulosucrase (IslA) from Leuconostoc citreum CW28

et al. 2008

View full text Add to dashboard Cite

BackgroundInulosucrase (IslA) from Leuconostoc citreum CW28 belongs to a new subfamily of multidomain fructosyltransferases (FTFs), containing additional domains from glucosyltransferases. It is not known what the function of the additional domains in this subfamily is.ResultsThrough construction of truncated versions we demonstrate that the acquired regions are involved in anchoring IslA to the cell wall; they also confer stability to the enzyme, generating a larger structure that affects its kinetic properties and reaction specificity, particularly the hydrolysis and transglycosylase ratio. The accessibility of larger molecules such as EDTA to the catalytic domain (where a Ca2+ binding site is located) is also affected as demonstrated by the requirement of 100 times higher EDTA concentrations to inactivate IslA with respect to the smallest truncated form.ConclusionThe C-terminal domain may have been acquired to anchor inulosucrase to the cell surface. Furthermore, the acquired domains in IslA interact with the catalytic core resulting in a new conformation that renders the enzyme more stable and switch the specificity from a hydrolytic to a transglycosylase mechanism. Based on these results, chimeric constructions may become a strategy to stabilize and modulate biocatalysts based on FTF activity.

show abstract

“…Contrary to Mur2, Mur1 does not possess amino acid repeats but rather contains a high “Proline plus Glycine plus Serine plus Threonine” content (33%), and it is has been suggested that, in certain Gram‐positive bacteria surface proteins, these domains rich in these specific residues are involved in cell wall binding. The proline–glycine residues are thought to be implicated in the interaction with the peptidoglycan–carbohydrate–teichoid acid matrix, whereas the serine–threonine residues are supposed to further stabilize this interaction [18].…”

Section: Discussionmentioning

confidence: 99%

The Bacillus thuringiensis phage GIL01 encodes two enzymes with peptidoglycan hydrolase activity

Verheust

Fornelos

Mahillon

2004

FEMS Microbiology Letters

View full text Add to dashboard Cite

Bacteriophage GIL01, infecting Bacillus thuringiensis serovar israelensis, possesses a linear dsDNA genome of 14,931 bp with proteins attached to its 5′ extremities and terminal inverted repeats at both ends. Viral particles are sensitive to organic solvents, suggesting that a lipid membrane is present in the capsid. All these characteristics are reminiscent of those found in members of the Tectiviridae family. Sequence analysis of GIL01 revealed the presence of two open reading frames (ORF25 and ORF30) encoding potential lytic enzymes, which were cloned and overexpressed in Escherichia coli. The muralytic activity of these two proteins, designated Mur1 and Mur2 respectively, was confirmed in situ using renaturing sodium dodecyl sulfate (SDS)–polyacrylamide gels containing bacterial cell wall preparations. While Mur2 degrading activity is limited to B. thuringiensis israelensis, Mur1 has a broader cleavage spectrum.

show abstract

Role of C-Terminal Domains in Surface Attachment of the Fructosyltransferase of Streptococcus salivariusATCC 25975

Cited by 5 publications

References 17 publications

Purification and enzymic properties of the fructosyltransferase of Streptococcus salivarius ATCC 25975

Purification and enzymic properties of the fructosyltransferase of Streptococcus salivarius ATCC 25975

Functional role of the additional domains in inulosucrase (IslA) from Leuconostoc citreum CW28

The Bacillus thuringiensis phage GIL01 encodes two enzymes with peptidoglycan hydrolase activity

Contact Info

Product

Resources

About