Production and purification of alternansucrase, a glucosyltransferase from Leuconostoc mesenteroides NRRL B-1355, for the synthesis of oligoalternans

López-Munguı́a, Agustı́n; Pelenc, V.; Remaud, M.; Biton, Jacques; Michel, J.-M.; Lang, Christine; Paul, F.; Monsan, Pierre

doi:10.1016/0141-0229(93)90120-q

Cited by 49 publications

(40 citation statements)

References 17 publications

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“…3C) and the N-terminal amino acid sequencing of the two forms showed that one form corresponds to the maturated ASR C-APY-del devoid of the 39-residue signal sequence and the second one lacks the next 102 residues, hence starting at amino acid 142. Both bands, cut from electrophoresis gels, catalyzed the formation of oligoalternans by acceptor reaction [3]. The specific activity of this purified preparation was of 160 U mg À1 .…”

Section: Contribution Of the Apy Motif To Asr Specificitymentioning

confidence: 98%

Construction of a fully active truncated alternansucrase partially deleted of its carboxy‐terminal domain

Joucla¹,

Pizzut

Monsan

et al. 2006

FEBS Letters

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Recombinant expression of the large alternansucrase (2057 amino acids) was hindered in E. coli due to poor enzyme solubility and protein degradation. The effects of deletions of the alternansucrase C-terminal CW-like and APY repeated motifs on enzyme solubility and specificity were investigated. A truncated variant deleted of the APY repeats but harboring four C-terminal CW-like repeats displayed a high specific activity and the same specificity of product synthesis as the native enzyme. It is more soluble and suffers less degradation than full length alternansucrase. Hence this truncated variant is a promising tool for the further structural and kinetic study of this interesting enzyme.

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Section: Contribution Of the Apy Motif To Asr Specificitymentioning

confidence: 98%

Construction of a fully active truncated alternansucrase partially deleted of its carboxy‐terminal domain

Joucla¹,

Pizzut

Monsan

et al. 2006

FEBS Letters

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“…Alternan represented about 30 to 50% of the water-soluble polysaccharide produced by cultures (13); it was purified from mixtures containing dextran by treatment with dextranase followed by alcohol precipitation (3), but yields were suboptimal because of losses to dextran synthesis. Alternansucrase, the enzyme responsible for alternan synthesis, has been partially purified, and its properties have been described (3,4,13,14,22).…”

mentioning

confidence: 99%

“…Alternan was estimated as the amount of polysaccharide resistant to hydrolysis by dextranase (3,13). Polysaccharide from the supernatant fraction of 100 ml of culture was precipitated by adding ethanol to a final concentration of 50% (vol/vol).…”

mentioning

confidence: 99%

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Glucosyltransferase Mutants of Leuconostoc mesenteroides NRRL B-1355

Smith

Zahnley

Goodman

1994

Appl Environ Microbiol

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“…Thermal stability of GTFs was studied after incubation at 45°C for 20 and 40 min in order to inactive DS. Alternansucrase is resistant to thermal inactivation (Lopez‐Munguia et al. 1993; Smith et al.…”

Section: Methodsmentioning

confidence: 99%

Gluco-oligosaccharides synthesized by glucosyltransferases from constitutive mutants of Leuconostoc mesenteroides strain Lm 28

et al. 2007

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Aims: To find different types of glucosyltransferases (GTFs) produced by Leuconostoc mesenteroides strain Lm 28 and its mutant forms, and to check the effectiveness of gluco‐oligosaccharide synthesis using maltose as the acceptor. Methods and Results: Constitutive mutants were obtained after chemical mutagenesis by ethyl methane sulfonate. Lm M281 produced more active GTFs than that obtained by the parental strain cultivated on sucrose. GTF from Lm M286 produced a resistant glucan, based on endo‐dextranase and amyloglucosidase hydrolysis. The extracellular enzymes from Lm M286 catalyse acceptor reactions and transfer the glucose unit from sucrose to maltose to produce gluco‐oligosaccharides (GOS). By increasing the sucrose/maltose ratio, it was possible to catalyse the synthesis of oligosaccharides of increasing degree of polymerization (DP). Conclusions: Different types of GTFs (dextransucrase, alternansucrase and levansucrase) were produced from new constitutive mutants of Leuc. mesenteroides. GTFs from Lm M286 can catalyse the acceptor reaction in the presence of maltose, leading to the synthesis of branched oligosaccharides. Significance and Impact of the Study: Conditions were optimized to synthesize GOS by using GTFs from Lm M286, with the aim of producing maximum quantities of branched‐chain oligosaccharides with DP 3–5. This would allow the use of the latter as prebiotics.

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Production and purification of alternansucrase, a glucosyltransferase from Leuconostoc mesenteroides NRRL B-1355, for the synthesis of oligoalternans

Cited by 49 publications

References 17 publications

Construction of a fully active truncated alternansucrase partially deleted of its carboxy‐terminal domain

Construction of a fully active truncated alternansucrase partially deleted of its carboxy‐terminal domain

Glucosyltransferase Mutants of Leuconostoc mesenteroides NRRL B-1355

Gluco-oligosaccharides synthesized by glucosyltransferases from constitutive mutants of Leuconostoc mesenteroides strain Lm 28

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