Cloning, sequence analysis and expression in Escherichia coli of a gene encoding an alginate lyase from Pseudomonas sp. OS-ALG-9

Maki, Hideshi; Mori, Atsutoshi; Fujiyama, Kazuhito; Kinoshita, Shinichi; Yoshida, Toshiomi

doi:10.1099/00221287-139-5-987

Cited by 45 publications

(29 citation statements)

References 29 publications

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“…OS-ALG-9 and 46% identity to a poly MG-specific alginate lyase AlyMG (AFC88009.1) from Stenotrophomonas maltophilia (25,26). Both AlyP and AlyMG are endo-type alginate lyases and have been assigned as members of the PL-6 family (25,26). As shown in the multiple sequence alignment, OalS6 had the same catalytic active and substrate-interacting sites conserved in the alginate lyases of the PL family 6 (Fig.…”

Section: Isolation and Identification Of Strain Kz7mentioning

confidence: 90%

“…These findings indicate that OalS6 may structurally different from other known oligoalginate lyases. Notably, although other alginate lyases in the PL-6 family also possess the chondroitinase-like domain, they are endo-type lyases, yielding unsaturated diand tri-saccharides as their main products (25,26). The oligoalginate lyases reported to date were predominantly polyM block preferred enzymes.…”

Section: Discussionmentioning

confidence: 99%

Section: Isolation and Identification Of Strain Kz7mentioning

confidence: 99%

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Cloning and characterization of the first polysaccharide lyase family 6 oligoalginate lyase from marineShewanellasp. Kz7

Li¹,

Wang²,

Han³

et al. 2015

J Biochem

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Section: Isolation and Identification Of Strain Kz7mentioning

confidence: 90%

Section: Discussionmentioning

confidence: 99%

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Cloning and characterization of the first polysaccharide lyase family 6 oligoalginate lyase from marineShewanellasp. Kz7

Li¹,

Wang²,

Han³

et al. 2015

J Biochem

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“…However, the deduced amino acid sequence revealed significant homology to a polysaccharide lyase, alginate lyase from Pseudomonas sp. (24), in the N-terminal region of both proteins. The ChnB protein might share structural similarities and an evolutionary relationship with the alginate lyase.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Expression in Escherichia coli of cslA and cslB , Genes Coding for the Chondroitin Sulfate-Degrading Enzymes Chondroitinase AC and Chondroitinase B, Respectively, from Flavobacterium heparinum

Tkalec¹,

Fink²,

Blain³

et al. 2000

Appl Environ Microbiol

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In medium supplemented with chondroitin sulfate, Flavobacterium heparinum synthesizes and exports two chondroitinases, chondroitinase AC (chondroitin AC lyase; EC 4.2.2.5) and chondroitinase B (chondroitin B lyase; no EC number), into its periplasmic space. Chondroitinase AC preferentially depolymerizes chondroitin sulfates A and C, whereas chondroitinase B degrades only dermatan sulfate (chondroitin sulfate B). The genes coding for both enzymes were isolated from F. heparinum and designated cslA (chondroitinase AC) and cslB (chondroitinase B). They were found to be separated by 5.5 kb on the chromosome of F. heparinum, transcribed in the same orientation, but not linked to any of the heparinase genes. In addition, the synthesis of both enzymes appeared to be coregulated. The cslA and cslB DNA sequences revealed open reading frames of 2,103 and 1,521 bp coding for peptides of 700 and 506 amino acid residues, respectively. Chondroitinase AC has a signal sequence of 22 residues, while chondroitinase B is composed of 25 residues. The mature forms of chondroitinases AC and B are comprised of 678 and 481 amino acid residues and have calculated molecular masses of 77,169 and 53,563 Da, respectively. Truncated cslA and cslB genes have been used to produce active, mature chondroitinases in the cytoplasm of Escherichia coli. Partially purified recombinant chondroitinases AC and B exhibit specific activities similar to those of chondroitinases AC and B from F. heparinum.

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“…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.…”

mentioning

confidence: 99%

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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Cloning, sequence analysis and expression in Escherichia coli of a gene encoding an alginate lyase from Pseudomonas sp. OS-ALG-9

Cited by 45 publications

References 29 publications

Cloning and characterization of the first polysaccharide lyase family 6 oligoalginate lyase from marineShewanellasp. Kz7

Cloning and characterization of the first polysaccharide lyase family 6 oligoalginate lyase from marineShewanellasp. Kz7

Isolation and Expression in Escherichia coli of cslA and cslB , Genes Coding for the Chondroitin Sulfate-Degrading Enzymes Chondroitinase AC and Chondroitinase B, Respectively, from Flavobacterium heparinum

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

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