Fine chemical structure analysis of oligosaccharides produced by an ulvan-lyase degradation of the water-soluble cell-wall polysaccharides from Ulva sp. (Ulvales, Chlorophyta)

Lahaye, Marc; Brunel, Magali; Bonnin, Estelle

doi:10.1016/s0008-6215(97)00270-x

Cited by 119 publications

(92 citation statements)

References 18 publications

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“…This targeted screening strategy enabled identification of a new ulvan lyase, LOR_107, with a protein sequence unrelated to the previously characterized N. ulvanivorans ulvan lyase (9). 1 H NMR analysis of the degradation end products, composed essentially of ⌬-R3S and ⌬-R3S-IdoUA-R3S, indicated that LOR_107 ulvan lyase cleaves specifically the ␤(134) glycoside bond between GlcUA and R3S residues according to a ␤-elimination mechanism. This substrate specificity is a unique feature, as the previously identified ulvan lyase in N. ulvanivorans cleaves both GlcUA and IdoUA residues by abstraction of the proton on the C5 position in both configurations, syn or anti, with regard to the hydroxyl group on C4 (9).…”

Section: Discussionmentioning

confidence: 99%

“…The ulvan lyase end products were purified and characterized by 1 H NMR according to Lahaye et al (1). The most abundant end products eluting at 63 min were disaccharides attributed to the 4-deoxy-L-threo-hex-4-enopyranosiduronic acid linked to sulfated rhamnose (⌬-R3S␣/␤, Fig.…”

Section: Table 2 Primers Used In This Studymentioning

confidence: 99%

“…GL2 (5), possess limited ulvanolytic activity. The first ulvan lyase activity was found in a marine bacterium by Lahaye et al (1), who employed the newly discovered enzyme extract to degrade ulvan for structural analysis. More recently, several bacterial strains capable of metabolizing ulvan were isolated from the feces of a sea slug, Aplysia punctate, fed with Ulva.…”

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New Family of Ulvan Lyases Identified in Three Isolates from the Alteromonadales Order

Kopel

Helbert

Belnik

et al. 2016

Journal of Biological Chemistry

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Ulvan is the main polysaccharide component of the Ulvales (green seaweed) cell wall. It is composed of disaccharide building blocks comprising 3-sulfated rhamnose linked to D-glucuronic acid (GlcUA), L-iduronic acid (IdoUA), or D-xylose (Xyl). The degradation of ulvan requires ulvan lyase, which catalyzes the endolytic cleavage of the glycoside bond between 3-sulfated rhamnose and uronic acid according to a ␤-elimination mechanism. The first characterized ulvan lyase was identified in Nonlabens ulvanivorans, an ulvanolytic bacterial isolate. In the current study, we have identified and biochemically characterized novel ulvan lyases from three Alteromonadales isolated bacteria. Two homologous ulvan lyases (long and short) were found in each of the bacterial genomes. The protein sequences have no homology to the previously reported ulvan lyases and therefore are the first representatives of a new family of polysaccharide lyases. The enzymes were heterologously expressed in Escherichia coli to determine their mode of action. The heterologous expressed enzymes were secreted into the milieu subsequent to their signal sequence cleavage. An endolytic mode of action was observed and studied using gel permeation chromatography and 1 H NMR. In contrast to N. ulvanivorans ulvan lyase, cleavage occurred specifically at the GlcUA residues. In light of the genomic context and modular structure of the ulvan lyase families identified to date, we propose that two ulvan degradation pathways evolved independently.

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Section: Discussionmentioning

confidence: 99%

Section: Table 2 Primers Used In This Studymentioning

confidence: 99%

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New Family of Ulvan Lyases Identified in Three Isolates from the Alteromonadales Order

Kopel

Helbert

Belnik

et al. 2016

Journal of Biological Chemistry

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“…In addition, the development of strategies that lead to the saccharification of ulvan, conversion from polymers into fermentable monosaccharides, requires the discovery of new enzymes because ulvan has no structural homolog in marine or land plants. An extracellular ulvan lyase has been isolated from a marine Gram-negative bacterium isolated from decomposing algae (4). Characterization of the bacteria was not reported, but the fractionated enzyme cleaves the ␤(134) linkage between Rha3S and GlcA leading to the production of unsaturated uronic acid (⌬, 4-deoxy-L-threohex-4-enopyranosiduronic acid, Fig.…”

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Ulvan Lyases Isolated from the Flavobacteria Persicivirga ulvanivorans Are the First Members of a New Polysaccharide Lyase Family

Collén¹,

Sassi²,

Rogniaux³

et al. 2011

Journal of Biological Chemistry

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Background: Ulvan, an anionic polysaccharide, is the most abundant component of the green algal cell wall. Results: The isolated ulvan lyases cleave the glycosidic linkage of ulvan between IduA/GlcA and Rha3S. The gene sequence has no homology in databases. Conclusion: Ulvan lyases are the first members of a new polysaccharide lyase family. Significance: Characterization of ulvan lyase is a first step toward understanding the degradation of ulvan.

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“…Oligosaccharides from a degree of polymerization of 2 to 3 or 5 can be obtained by polysaccharide degradation with lyases (3,20,40). To date, lyase activities on various polymers have been described as alginate (16,32,35), gellan (41), hyaluronan (39), ulvan (25), and xanthan (15). The tested lyases were found in many organisms such as marine gastropods (19) or fungi (36,45) as well as in many bacteria (14,15,31) and bacteriophages (1,4).…”

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Purification and Properties of a Glucuronan Lyase from Sinorhizobium meliloti M5N1CS (NCIMB 40472)

Costa

Michaud

Petit

et al. 2001

Appl Environ Microbiol

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A glucuronan lyase extracted from Sinorhizobium meliloti strain M5N1CS was purified to homogeneity by anion-exchange chromatography. The purified enzyme corresponds to a monomer with a molecular mass of 20 kDa and a pI of 4.9. A specific activity was found only for polyglucuronates leading to the production of 4,5-unsaturated oligoglucuronates. The enzyme activity was optimal at pH 6.5 and 50°C. Zn 2؉ , Cu 2؉ , and Hg 2؉(1 mM) inhibited the enzyme activity. No homology of the enzyme N-terminal amino acid sequence was found with any of the previously published protein sequences. This enzyme purified from S. meliloti strain M5N1CS corresponding to a new lyase was classified as an endopolyglucuronate lyase.Bacterial polysaccharides as well as plant polysaccharides can be degraded by specific glycosidases corresponding either to hydrolases or to lyases. Among the polyanionic polymers, some of them (i.e., alginate) are exclusively degraded by lyases (35,40,42). The lyase cleaving mechanism consists of a ␤-elimination, in which a general base-catalyzed abstraction of the proton at C-5 of a uronic acid occurs. A transfer of electrons from the carboxyl group to form a double bond between C-4 and C-5 results in the elimination of the 4-O-glycosidic bond and in the formation of 4-deoxy-L-erythro-hex-4-enopyranosyluronic acid. This reaction leads to the formation of an unsaturated uronate at the newly generated, nonreducing end. Oligosaccharides from a degree of polymerization of 2 to 3 or 5 can be obtained by polysaccharide degradation with lyases (3,20,40). To date, lyase activities on various polymers have been described as alginate (16,32,35), gellan (41), hyaluronan (39), ulvan (25), and xanthan (15). The tested lyases were found in many organisms such as marine gastropods (19) or fungi (36, 45) as well as in many bacteria (14,15,31) and bacteriophages (1, 4). The enzyme localization in the producing organisms may be in the cytosol (28, 36) and the periplasmic space (22), as for alginate lyase from Azotobacter species, or in extracellular fractions, as for a Bacillus circulans lyase (26). Lyases present possible applications in the medical field, such as, for example, the alginate lyase, which promotes diffusion of antibiotics through extracellular polymers produced by the pathogenic Pseudomonas sp. (17). Due to potential applications, studies concerning polysaccharide lyases have been increasing.The Sinorhizobium meliloti mutant strain M5N1CS (NCIMB 40472) (7), which induces the formation of effective nodules on alfalfa roots (12), produces a glucuronan (6, 18) corresponding to high-molecular-weight (HMW) and low-molecular-weight (LMW) (134)-␤-D-polyglucuronic acid partially acetylated at the C-2 and/or C-3 position. The detection of LMW glucuronan containing a 4-5 unsaturated glucuronic residue at the nonreducing end (29, 30) led us to suspect a polymer degradation by a glucuronan lyase.In order to produce oligoglucuronans, a lyase degrading various acetylated and nonacetylated glucuronans was purified and character...

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Fine chemical structure analysis of oligosaccharides produced by an ulvan-lyase degradation of the water-soluble cell-wall polysaccharides from Ulva sp. (Ulvales, Chlorophyta)

Cited by 119 publications

References 18 publications

New Family of Ulvan Lyases Identified in Three Isolates from the Alteromonadales Order

New Family of Ulvan Lyases Identified in Three Isolates from the Alteromonadales Order

Ulvan Lyases Isolated from the Flavobacteria Persicivirga ulvanivorans Are the First Members of a New Polysaccharide Lyase Family

Purification and Properties of a Glucuronan Lyase from Sinorhizobium meliloti M5N1CS (NCIMB 40472)

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