A Partially Pyruvated Carrageenan from Hawaiian <i>Grateloupia filicina </i>(Cryptonemiales, Rhodophyta)

Zablackis, Earl; Pérez, Jose Antonio Pérez

doi:10.1515/botm.1990.33.3.273

Cited by 35 publications

(19 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…B, time course of the hydrolysis of -carrageenan with Z. galactanovorans -carrageenase (0.2 UA/mg of -carrageenan). At intervals, aliquots from the reaction mixture were boiled and analyzed by carbohydrate-PAGE (30). The lane marked T1 was loaded with purified -neocarratetraose (DP2).…”

Section: Resultsmentioning

confidence: 99%

ι-Carrageenases Constitute a Novel Family of Glycoside Hydrolases, Unrelated to That of κ-Carrageenases

Barbeyron¹,

Michel²,

Potin³

et al. 2000

Journal of Biological Chemistry

123

111

View full text Add to dashboard Cite

-Carrageenases are polysaccharide hydrolases that cleave the ␤-1,4 linkages between the D-galactose-4-sulfate and 3,6-anhydro-D-galactose-2-sulfate residues in the red algal galactans known as -carrageenans. We report here on the purification of -carrageenase activity from the marine bacterium Zobellia galactanovorans and on the characterization of -carrageenase structural genes. Genomic libraries from this latter bacterium as well as from Alteromonas fortis were functionally screened for the presence of -carrageenase ؉ clones. The Z. galactanovorans and A. fortis -carrageenase genes encode homologous proteins of 53.4 and 54.8 kDa, respectively. Based on hydrophobic cluster analysis and on the 1 H NMR monitoring of the products of the overexpressed A. fortis -carrageenase, these enzymes appear to form a new family of glycoside hydrolases, unrelated to that of -carrageenases and with an inverting mechanism of hydrolysis. They both feature a 45-amino acid-long N-terminal segment with sequence similarity to the N-terminal region of several other polysaccharidases. In those for which a three-dimensional structure is available, this conspicuous segment, also deemed "glycanase motif" (Chua, J. E. H., Manning, P. A., and Morona, R. (1999) Microbiology (Reading) 145, 1649 -1659), corresponds to a strand-helix-strand "cap" that covers the N-terminal end of a common, right-handed ␤-helical fold.

show abstract

Section: Resultsmentioning

confidence: 99%

ι-Carrageenases Constitute a Novel Family of Glycoside Hydrolases, Unrelated to That of κ-Carrageenases

Barbeyron¹,

Michel²,

Potin³

et al. 2000

Journal of Biological Chemistry

123

111

View full text Add to dashboard Cite

show abstract

“…The λ-carrageenase solution (500 µg/ml) was diluted 50 times in a buffer containing 0.5 % λ-carrageenan in 0.1 M NaNO 3 (pH 7.5). The reaction medium was incubated for 96 h at 30 • C or for 2 weeks at 20 • C. The release of λ-oligosaccharides was visualized using C/PAGE (carbohydrate/PAGE) [30].…”

Section: Expression Of Recombinant λ-Carrageenasementioning

confidence: 99%

“…Since DP2 was eluted with salts and other small molecules, it could not be purified by sizeexclusion chromatography. Therefore fluorescent DP2 was purified by C/PAGE [27% (w/v)] [30]. The fluorescent band corresponding to the labelled DP2 was excised, ground and allowed to diffuse in a small amount of water at 4 • C. After 24 h, the fluorescent DP2 was recovered, freeze-dried and stored at 4 • C.…”

Section: Synthesis and Enzymatic Digestion Of Fluorescent Oligosacchamentioning

confidence: 99%

See 1 more Smart Citation

Degradation of λ-carrageenan by Pseudoalteromonas carrageenovora λ-carrageenase: a new family of glycoside hydrolases unrelated to κ- and ι-carrageenases

Guibet¹,

Colin²,

Barbeyron³

et al. 2007

Biochem. J.

View full text Add to dashboard Cite

Carrageenans are sulfated galactans found in the cell walls of red seaweeds. They are classified according to the number and the position of sulfate ester groups. lambda-Carrageenan is the most sulfated carrageenan and carries at least three sulfates per disaccharide unit. The sole known depolymerizing enzyme of lambda-carrageenan, the lambda-carrageenase from Pseudoalteromonas carrageenovora, has been purified, cloned and sequenced. Sequence analyses have revealed that the lambda-carrageenase, referred to as CglA, is the first member of a new family of GHs (glycoside hydrolases), which is unrelated to families GH16, that contains kappa-carrageenases, and GH82, that contains iota-carrageenases. This large enzyme (105 kDa) features a low-complexity region, suggesting the presence of a linker connecting at least two independent modules. The N-terminal region is predicted to fold as a beta-propeller. The main degradation products have been purified and characterized as neo-lambda-carratetraose [DP (degree of polymerization) 4] and neo-lambda-carrahexaose (DP6), indicating that CglA hydrolyses the beta-(1-->4) linkage of lambda-carrageenan. LC-MALLS (liquid chromatography-multi-angle laser light scattering) and (1)H-NMR monitoring of the enzymatic degradation of lambda-carrageenan indicate that CglA proceeds according to an endolytic mode of action and a mechanism of inversion of the anomeric configuration. Using 2-aminoacridone-labelled neo-lambda-carrabiose oligosaccharides, in the present study we demonstrate that the active site of CglA comprises at least 8 subsites (-4 to +4) and that a DP6 oligosaccharide binds in the subsites -4 to +2 and can be hydrolysed into DP4 and DP2.

show abstract

“…5 l were electrophoresed through a 6% (w/v) stacking and a 28% running, 0.75-mm thick, polyacrylamide gel in 50 mM Tris-HCl, 1 mM EDTA buffer, pH 8.7. Gels were stained with Alcian blue 0.5% (w/v) followed by silver nitrate 0.4% (w/v) (73,74 Prior to an enzymatic reaction, a reference spectrum of the substrate was acquired. The sample was then removed from the probe; the enzyme was added at the proper concentration, and the tube was put back into the probe.…”

mentioning

confidence: 99%

Comparative Characterization of Two Marine Alginate Lyases from Zobellia galactanivorans Reveals Distinct Modes of Action and Exquisite Adaptation to Their Natural Substrate

Thomas

Lundqvist

Jam

et al. 2013

Journal of Biological Chemistry

175

207

View full text Add to dashboard Cite

Background: Alginolytic systems from marine bacteria are crucial for algal biomass conversion, yet their molecular mechanisms remain poorly understood. Results: Structural and biochemical characterization of two paralogous marine alginate lyases highlights details on complementary roles and differences with terrestrial enzymes. Conclusion: Bacterial alginolytic enzymes are specifically adapted to the unique characteristics of the natural substrate. Significance: Marine microbes evolved complex degradation systems targeting habitat-specific polysaccharides.

show abstract

A Partially Pyruvated Carrageenan from Hawaiian Grateloupia filicina (Cryptonemiales, Rhodophyta)

Cited by 35 publications

References 17 publications

ι-Carrageenases Constitute a Novel Family of Glycoside Hydrolases, Unrelated to That of κ-Carrageenases

ι-Carrageenases Constitute a Novel Family of Glycoside Hydrolases, Unrelated to That of κ-Carrageenases

Degradation of λ-carrageenan by Pseudoalteromonas carrageenovora λ-carrageenase: a new family of glycoside hydrolases unrelated to κ- and ι-carrageenases

Comparative Characterization of Two Marine Alginate Lyases from Zobellia galactanivorans Reveals Distinct Modes of Action and Exquisite Adaptation to Their Natural Substrate

Contact Info

Product

Resources

About