Bacillus subtilis CwlQ (previous YjbJ) is a bifunctional enzyme exhibiting muramidase and soluble-lytic transglycosylase activities

Sudiarta, I Putu; Fukushima, Tatsuya; Sekiguchi, Junichi

doi:10.1016/j.bbrc.2010.07.001

Cited by 19 publications

(14 citation statements)

References 19 publications

(1 reference statement)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In summary, we identified the typical anhydromuropeptides resulting from LT activity in addition to several reaction products containing a reducing end (compounds corresponding to peaks A, D, F, and G). Similarly, PG fragments with a terminal reducing end have been previously reported to be reaction products of Bacillus subtilis and E. coli LTs, especially for enzymes exhibiting low activity in vitro ( 46 – 48 ). These products are proposed to be formed when the transient oxocarbenium intermediate entraps a water molecule instead of the C6-hydroxyl group of NAM, giving rise to hydrolytic cleavage of the NAM-NAG glycosidic bond (muramidase activity) rather than to nonhydrolytic cleavage resulting in anhydroNAM formation (LT activity).…”

Section: Resultsmentioning

confidence: 91%

Cwp19 Is a Novel Lytic Transglycosylase Involved in Stationary-Phase Autolysis Resulting in Toxin Release in Clostridium difficile

Wydau-Dematteis

Meouche

Courtin

et al. 2018

mBio

View full text Add to dashboard Cite

Clostridium difficile is the major etiologic agent of antibiotic-associated intestinal disease. Pathogenesis of C. difficile is mainly attributed to the production and secretion of toxins A and B. Unlike most clostridial toxins, toxins A and B have no signal peptide, and they are therefore secreted by unusual mechanisms involving the holin-like TcdE protein and/or autolysis. In this study, we characterized the cell surface protein Cwp19, a newly identified peptidoglycan-degrading enzyme containing a novel catalytic domain. We purified a recombinant His6-tagged Cwp19 protein and showed that it has lytic transglycosylase activity. Moreover, we observed that Cwp19 is involved in cell autolysis and that a C. difficile cwp19 mutant exhibited delayed autolysis in stationary phase compared to the wild type when bacteria were grown in brain heart infusion (BHI) medium. Wild-type cell autolysis is correlated to strong alterations of cell wall thickness and integrity and to release of cytoplasmic material. Furthermore, we demonstrated that toxins were released into the extracellular medium as a result of Cwp19-induced autolysis when cells were grown in BHI medium. In contrast, Cwp19 did not induce autolysis or toxin release when cells were grown in tryptone-yeast extract (TY) medium. These data provide evidence for the first time that TcdE and bacteriolysis are coexisting mechanisms for toxin release, with their relative contributions in vitro depending on growth conditions. Thus, Cwp19 is an important surface protein involved in autolysis of vegetative cells of C. difficile that mediates the release of the toxins from the cell cytosol in response to specific environment conditions.

show abstract

Section: Resultsmentioning

confidence: 91%

Cwp19 Is a Novel Lytic Transglycosylase Involved in Stationary-Phase Autolysis Resulting in Toxin Release in Clostridium difficile

Wydau-Dematteis

Meouche

Courtin

et al. 2018

mBio

View full text Add to dashboard Cite

show abstract

“…those containing 18 O) were observed in the soluble reaction products of SltF (Table 2). This observation is not unexpected given the reported secondary (minor) muramidase-like activity of LTs, such as MltD, Slt70, MltF from E. coli (36), and CwlQ from Bacillus subtilis (37).…”

Section: Resultsmentioning

confidence: 53%

Modulation of the Lytic Activity of the Dedicated Autolysin for Flagellum Formation SltF by Flagellar Rod Proteins FlgB and FlgF

et al. 2016

View full text Add to dashboard Cite

SltF was identified previously as an autolysin required for the assembly of flagella in the alphaproteobacteria, but the nature of its peptidoglycan lytic activity remained unknown. Sequence alignment analyses suggest that it could function as either a muramidase, lytic transglycosylase, or ␤-N-acetylglucosaminidase. Recombinant SltF from Rhodobacter sphaeroides was purified to apparent homogeneity, and it was demonstrated to function as a lytic transglycosylase based on enzymatic assays involving mass spectrometric analyses. Circular dichroism (CD) analysis determined that it is composed of 83.4% ␣-structure and 1.48% ␤-structure and thus is similar to family 1A lytic transglycosylases. However, alignment of apparent SltF homologs identified in the genome database defined a new subfamily of the family 1 lytic transglycosylases. SltF was demonstrated to be endo-acting, cleaving within chains of peptidoglycan, with optimal activity at pH 7.0. Its activity is modulated by two flagellar rod proteins, FlgB and FlgF: FlgB both stabilizes and stimulates SltF activity, while FlgF inhibits it. Invariant Glu57 was confirmed as the sole catalytic acid/base residue of SltF. IMPORTANCEThe bacterial flagellum is comprised of a basal body, hook, and helical filament, which are connected by a rod structure. With a diameter of approximately 4 nm, the rod is larger than the estimated pore size within the peptidoglycan sacculus, and hence its insertion requires the localized and controlled lysis of this essential cell wall component. In many beta-and gammaproteobacteria, this lysis is catalyzed by the ␤-N-acetylglucosaminidase domain of FlgJ. However, FlgJ of the alphaproteobacteria lacks this activity and instead it recruits a separate enzyme, SltF, for this purpose. In this study, we demonstrate that SltF functions as a newly identified class of lytic transglycosylases and that its autolytic activity is uniquely modulated by two rod proteins, FlgB and FlgF.T he photosynthetic bacterium Rhodobacter sphaeroides is motile through the use of a single, subpolar flagellum (1). This locomotive organelle is tightly regulated and comprised of approximately 25 different proteins arranged into three major substructures: a basal body, hook, and thin helical filament. The basal body spans the bacterial cell envelope and contains the motor, a flagellum-specific type III export apparatus and at least four ringlike structures which are all connected by a filamentous rod (2). The rod assembles into a proximal rod that lies between the MS ring and the cell wall, which is composed of nine subunits of FliE (3) and six subunits of FlgB, FlgC, and FlgF (4), as well as a distal rod that is composed of 26 subunits of FlgG (5).During flagellum assembly, extensive modifications need to occur to the peptidoglycan (PG) sacculus to accommodate the insertion of the secretion apparatus, as well as to stabilize the function of this system by acting as an assembly scaffold (6, 7). PG is a heteropolymer of glycan strands and peptide chains forming a rig...

show abstract

“…C, RP-HPLC of the reduced fragments of mutanolysin digests. Deacetylated or non-deacetylated reduced-4S2P material was digested with a muramidase, mutanolysin, followed by reduction with 2 and 3) is unknown, but it may reflect the difference of ␣ and ␤ anomers at the reducing end as described previously (32). Peak 1 in Fig.…”

Section: Confirmation Of Murnac Deacetylation Activity Of Pdac With Mmentioning

confidence: 99%

Identification and Characterization of a Novel Polysaccharide Deacetylase C (PdaC) from Bacillus subtilis

Kobayashi

Sudiarta

Kodama

et al. 2012

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Bacillus subtilis CwlQ (previous YjbJ) is a bifunctional enzyme exhibiting muramidase and soluble-lytic transglycosylase activities

Cited by 19 publications

References 19 publications

Cwp19 Is a Novel Lytic Transglycosylase Involved in Stationary-Phase Autolysis Resulting in Toxin Release in Clostridium difficile

Cwp19 Is a Novel Lytic Transglycosylase Involved in Stationary-Phase Autolysis Resulting in Toxin Release in Clostridium difficile

Modulation of the Lytic Activity of the Dedicated Autolysin for Flagellum Formation SltF by Flagellar Rod Proteins FlgB and FlgF

Identification and Characterization of a Novel Polysaccharide Deacetylase C (PdaC) from Bacillus subtilis

Contact Info

Product

Resources

About