Factors affecting α,‐1,2 glucooligosaccharide synthesis by <i>Leuconostoc mesenteroides</i> NRRL B‐1299 dextransucrase

Dols‐Lafargue, Marguerite; Willemot, René‐Marc; Monsan, Pierre; Remaud‐Siméon, Magali

doi:10.1002/bit.1141

Cited by 21 publications

(11 citation statements)

References 23 publications

(27 reference statements)

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“…Koepsell et al (89) observed that in the presence of sucrose and saccharide acceptor substrates such as maltose, isomaltose, and O-␣-methylglucoside, GS enzymes shift from glucan synthesis towards the synthesis of oligosaccharides (the acceptor reaction). Most acceptor reaction studies have been performed using saccharides (5,42,43,53,94,99) or saccharide derivatives as substrates (31,36,150). However, aromatic compounds (e.g., catechine) and salicyl alcohol have also been shown to act as acceptor substrates (114,221).…”

Section: Reactions Catalyzed and Glucan Product Synthesismentioning

confidence: 99%

Structure-Function Relationships of Glucansucrase and Fructansucrase Enzymes from Lactic Acid Bacteria

Hijum

Kralj

Ozimek

et al. 2006

Microbiol Mol Biol Rev

393

315

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SUMMARY Lactic acid bacteria (LAB) employ sucrase-type enzymes to convert sucrose into homopolysaccharides consisting of either glucosyl units (glucans) or fructosyl units (fructans). The enzymes involved are labeled glucansucrases (GS) and fructansucrases (FS), respectively. The available molecular, biochemical, and structural information on sucrase genes and enzymes from various LAB and their fructan and α-glucan products is reviewed. The GS and FS enzymes are both glycoside hydrolase enzymes that act on the same substrate (sucrose) and catalyze (retaining) transglycosylation reactions that result in polysaccharide formation, but they possess completely different protein structures. GS enzymes (family GH70) are large multidomain proteins that occur exclusively in LAB. Their catalytic domain displays clear secondary-structure similarity with α-amylase enzymes (family GH13), with a predicted permuted (β/α)8 barrel structure for which detailed structural and mechanistic information is available. Emphasis now is on identification of residues and regions important for GS enzyme activity and product specificity (synthesis of α-glucans differing in glycosidic linkage type, degree and type of branching, glucan molecular mass, and solubility). FS enzymes (family GH68) occur in both gram-negative and gram-positive bacteria and synthesize β-fructan polymers with either β-(2→6) (inulin) or β-(2→1) (levan) glycosidic bonds. Recently, the first high-resolution three-dimensional structures have become available for FS (levansucrase) proteins, revealing a rare five-bladed β-propeller structure with a deep, negatively charged central pocket. Although these structures have provided detailed mechanistic insights, the structural features in FS enzymes dictating the synthesis of either β-(2→6) or β-(2→1) linkages, degree and type of branching, and fructan molecular mass remain to be identified.

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Section: Reactions Catalyzed and Glucan Product Synthesismentioning

confidence: 99%

Structure-Function Relationships of Glucansucrase and Fructansucrase Enzymes from Lactic Acid Bacteria

Hijum

Kralj

Ozimek

et al. 2006

Microbiol Mol Biol Rev

393

315

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“…As indicated by the effectiveness factor ( maltose < 1), maltose consumption was slower with immobilized IGT; dextran yield was higher, even though the final residual maltose concentration [M] f was higher with the immobilized IGT (Table IV). With the free enzyme, dextran synthesis was observed only when maltose was exhausted (Dols-Lafargue et al, 2001). …”

Section: Performance Of the Immobilized Dextransucrase During Batch Ementioning

confidence: 95%

“…These properties, which result from the presence of ␣-1,2 linkages, render these GOS very attractive for applications in nutrition, cosmetics, or pharmaceuticals (Monsan and Paul, 1995). In a companion article, we describe the conditions with free NRRL B-1299 dextransucrase that led to a total GOS yield of 88% with more than 65% of the GOS bearing an ␣-1,2 linkage (Dols-Lafargue et al, 2001). But, to be economically feasible on the industrial scale, continuous ␣,1-2 GOS synthesis needs an immobilized catalyst.…”

Section: Introductionmentioning

confidence: 98%

Reactor optimization for α‐1,2 glucooligosaccharide synthesis by immobilized dextransucrase

Dols‐Lafargue

Willemot

Monsan

et al. 2001

Biotech & Bioengineering

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The immobilization of dextransucrase in Ca-alginate beads relies on the close association between dextran polymer and dextransucrase. However, high amounts of dextran in the enzyme preparation drastically limit the specific activity of the immobilized enzyme (4 U/mL of alginate beads). Moreover, even in the absence of diffusion limitation at the batch conditions used, the enzyme behavior is modified by entrapment so that the dextran yield increases and the alpha-1,2 glucooligosaccharides (GOS) are produced with a lower yield (46.6% instead of 56.7%) and have a lower mean degree of polymerization than with the free dextransucrase. When the immobilized catalyst is used in a continuous reaction, the reactor flow rate necessary to obtain high conversion of the substrates is very low, leading to external diffusion resistance. As a result, dextran synthesis is even higher than in the batch reaction, and its accumulation within the alginate beads limits the operational stability of the catalyst and decreases glucooligosaccharide yield and productivity. This effect can be limited by using reactor columns with length to diameter ratio > or =20, and by optimizing the substrate concentrations in the feed solution: the best productivity obtained was 3.74 g. U(-1). h(-1), with an alpha-1,2 GOS yield of 36%.

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“…In that case, successive glucopyranosyl transfers to the acceptor molecule lead to the formation of oligosaccharide series. Thus, using glucansucrases of distinctive specificities in the presence of different acceptors gives access to a large variety of oligosaccharide structures, as e.g., (i) leucrose (5-a-D-glucopyranosyl-D-fructopyranose, isomer of sucrose) [15], (ii) dextrans of controlled molecular weight [16], (iii) a-glucosylated cellobiose [17], and (iv) gluco-oligosaccharides with a1?2 glucosidic bonds produced on the industrial scale (50 t/year) [18,19].…”

Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis analysis of glucooligosaccharide regioisomers

Joucla¹,

Brando

Remaud‐Siméon³

et al. 2004

Electrophoresis

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Complex gluco-oligosaccharide mixtures of two regioisomer series were successfully separated by CE. The gluco-oligosaccharide series were synthesized, employing a dextransucrase from Leuconostoc mesenteroides NRRL B-512F, by successive glucopyranosyl transfers from sucrose to the acceptor glucose or maltose. The glucosyl transfer to both acceptors, occurring through the formation of alpha1-->6 linkages, differed for the two series only in the glucosidic bond to the reducing end namely alpha1-->6 or alpha1-->4 bond for glucose or maltose acceptor, respectively. Thus, the combination of the two series results in mixed pairs of gluco-oligosaccharide regioisomers with different degrees of polymerization (DP). These regioisomer series were first derivatized by reductive amination with 9-aminopyrene-1,4,6-trisulfonate (APTS). Under acidic conditions using triethyl ammonium acetate as electrolyte, the APTS-gluco-oligosaccharides of each series were separated enabling unambiguous size determination by coupling CE to electrospray-mass spectrometry. However, neither these acidic conditions nor alkaline buffer systems could be adapted for the separation of the gluco-oligosaccharide regioisomers arising from the two combined series. By contrast, increased resolution was observed in an alkaline borate buffer, using differential complexation of the regioisomers with the borate anions. Such conditions were also successfully applied to the separation of glucodisaccharide regioisomers composed of alpha1-->2, alpha1-->3, alpha1-->4, and alpha1-->6 linkages commonly synthesized by glucansucrase enzymes.

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Factors affecting α,‐1,2 glucooligosaccharide synthesis by Leuconostoc mesenteroides NRRL B‐1299 dextransucrase

Cited by 21 publications

References 23 publications

Structure-Function Relationships of Glucansucrase and Fructansucrase Enzymes from Lactic Acid Bacteria

Structure-Function Relationships of Glucansucrase and Fructansucrase Enzymes from Lactic Acid Bacteria

Reactor optimization for α‐1,2 glucooligosaccharide synthesis by immobilized dextransucrase

Capillary electrophoresis analysis of glucooligosaccharide regioisomers

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