Structural characterization ofAcetobacter diazotropicus levansucrase by matrix-assisted laser desorption/ionization mass spectrometry: identification of an N-terminal blocking group and a free-thiol cysteine residue

Betancourt, Lázaro; Takao, Toshifumi; Hernández, Lázaro; Padrón, Gabrìel; Shimonishi, Yasutsugu

doi:10.1002/(sici)1096-9888(199903)34:3<169::aid-jms780>3.0.co;2-4

Cited by 11 publications

(9 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…diazotrophicus (PDB:1W18) stabilizes the protein fold by connecting the extended loop between β‐strands IIIB and IIIC with the insertion located between blades III and IV (Fig. A) (Betancourt et al ., ; Martínez‐Fleites et al ., ). An equivalent pair of Cys residues is strictly conserved in all T2‐LSs from different origins (Fig.…”

Section: Resultsmentioning

confidence: 97%

Acetic acid bacteria encode two levansucrase types of different ecological relationship

Jakob

Quintero

Musacchio

et al. 2019

Environmental Microbiology

View full text Add to dashboard Cite

Summary Acetic acid bacteria (AAB) are associated with plants and insects. Determinants for the targeting and occupation of these widely different environments are unknown. However, most of these natural habitats share plant‐derived sucrose, which can be metabolized by some AAB via polyfructose building levansucrases (LS) known to be involved in biofilm formation. Here, we propose two LS types (T) encoded by AAB as determinants for habitat selection, which emerged from vertical (T1) and horizontal (T2) lines of evolution and differ in their genetic organization, structural features and secretion mechanism, as well as their occurrence in proteobacteria. T1‐LS are secreted by plant‐pathogenic α‐ and γ‐proteobacteria, while T2‐LS genes are common in diazotrophic, plant‐growth‐promoting α‐, β‐ and γ‐proteobacteria. This knowledge may be exploited for a better understanding of microbial ecology, plant health and biofilm formation by sucrase‐secreting proteobacteria in eukaryotic hosts.

show abstract

Section: Resultsmentioning

confidence: 97%

Acetic acid bacteria encode two levansucrase types of different ecological relationship

Jakob

Quintero

Musacchio

et al. 2019

Environmental Microbiology

View full text Add to dashboard Cite

show abstract

“…Capital letters correspond to proposed components of the active center. Mature form of the enzyme50, 51 is shown by box. Bracket indicates a disulfide bond.…”

Section: Resultsmentioning

confidence: 99%

?-Fructosidase superfamily: Homology with some ?-L-arabinases and ?-D-xylosidases

Naumoff

2000

Proteins

View full text Add to dashboard Cite

“…The mature protein thus comprises a 553-residue polypeptide (numbering 31-584) with an N-terminal pyroglutamate. LsdA contains three cysteine residues (Cys 157 , Cys 339 and Cys 395 ), with Cys 339 and Cys 395 engaged in a disulphide bridge [31].…”

Section: Structure Of Gd Levansucrase Lsdamentioning

confidence: 99%

Crystal structure of levansucrase from the Gram-negative bacterium Gluconacetobacter diazotrophicus

Martinez-Fleites

Ortiz-Lombardı́a

Pons

et al. 2005

Biochemical Journal

143

125

View full text Add to dashboard Cite

The endophytic Gram-negative bacterium Gluconacetobacter diazotrophicus SRT4 secretes a constitutively expressed levansucrase (LsdA, EC 2.4.1.10), which converts sucrose into fructooligosaccharides and levan. The enzyme is included in GH (glycoside hydrolase) family 68 of the sequence-based classification of glycosidases. The three-dimensional structure of LsdA has been determined by X-ray crystallography at a resolution of 2.5 A (1 A=0.1 nm). The structure was solved by molecular replacement using the homologous Bacillus subtilis (Bs) levansucrase (Protein Data Bank accession code 1OYG) as a search model. LsdA displays a five-bladed beta-propeller architecture, where the catalytic residues that are responsible for sucrose hydrolysis are perfectly superimposable with the equivalent residues of the Bs homologue. The comparison of both structures, the mutagenesis data and the analysis of GH68 family multiple sequences alignment show a strong conservation of the sucrose hydrolytic machinery among levansucrases and also a structural equivalence of the Bs levansucrase Ca2+-binding site to the LsdA Cys339-Cys395 disulphide bridge, suggesting similar fold-stabilizing roles. Despite the strong conservation of the sucrose-recognition site observed in LsdA, Bs levansucrase and GH32 family Thermotoga maritima invertase, structural differences appear around residues involved in the transfructosylation reaction.

show abstract

Structural characterization ofAcetobacter diazotropicus levansucrase by matrix-assisted laser desorption/ionization mass spectrometry: identification of an N-terminal blocking group and a free-thiol cysteine residue

Cited by 11 publications

References 16 publications

Acetic acid bacteria encode two levansucrase types of different ecological relationship

Acetic acid bacteria encode two levansucrase types of different ecological relationship

?-Fructosidase superfamily: Homology with some ?-L-arabinases and ?-D-xylosidases

Crystal structure of levansucrase from the Gram-negative bacterium Gluconacetobacter diazotrophicus

Contact Info

Product

Resources

About