Substrate complexation and aggregation influence the cyclodextrin glycosyltransferase (CGTase) catalyzed synthesis of alkyl glycosides

Zehentgruber, Daniela; Lundemo, Pontus; Svensson, David; Adlercreutz, Patrick

doi:10.1016/j.jbiotec.2011.06.011

Cited by 9 publications

(5 citation statements)

References 13 publications

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“…Previous studies have indicated that CD and AGly can form complexes and that complexation influences the kinetics of CGTase-catalyzed reactions involving AGly and CD substrates. 30 To elucidate these mechanisms in detail, the formation of inclusion complexes between αCD and C8G1 was studied separately under the conditions used in the enzymatic synthesis reactions, by isothermal titration calorimetry (ITC). The results showed that the 1:1 complex (αCD:C8G1) was the dominating species.…”

Section: Disproportionation: Aggregation Solubility and Complexation ...mentioning

confidence: 99%

“…Promising results were obtained by Zehentgruber at al. 30 in elongating the maltose headgroup of dodecyl-β-Dmaltoside (C12G2) by applying an excess amount of αCD. This system combines the complexation process with CD as solubilization agent and being an actual substrate.…”

Section: Introductionmentioning

confidence: 99%

“…So far, mainly alkyl maltosides have been used as starting material because they possess better solubility and reactivity than AGlu’s. Promising results were obtained by Zehentgruber at al . in elongating the maltose headgroup of dodecyl-β- d -maltoside (C12G2) by applying an excess amount of αCD.…”

Section: Introductionmentioning

confidence: 99%

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Hydrophobic Complexation Promotes Enzymatic Surfactant Synthesis from Alkyl Glucoside/Cyclodextrin Mixtures

Börner¹,

Roger²,

Adlercreutz

2014

ACS Catal.

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The unique ability of cyclodextrin glycosyltransferase to form and utilize the cyclic maltooligosaccharide cyclodextrin (CD) makes this enzyme an attractive catalyst for the synthesis of alkyl glycosides. Here, we characterize the sugar headgroup elongation of alkyl glucosides (acceptor) via two transglycosylation reactions from either a linear (maltohexose) or a cyclic (CD) glycosyl donor. Inclusion complex formation overcomes both poor substrate solubility and aggregation. We have used pure alkyl glucosides and αCD as model compounds. The complex between CD and alkyl glucoside was efficiently used as a substrate. Kinetic and thermodynamic measurements allow the prediction of the optimal synthesis conditions. This optimum corresponds to the transition between a donor-limiting and an acceptor-limiting regime. The resulting rational design should lead to the practical development of a cost-efficient industrial synthesis. Our findings with respect to the importance of complexation should also readily apply to other enzymatic systems.

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Section: Disproportionation: Aggregation Solubility and Complexation ...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Hydrophobic Complexation Promotes Enzymatic Surfactant Synthesis from Alkyl Glucoside/Cyclodextrin Mixtures

Börner¹,

Roger²,

Adlercreutz

2014

ACS Catal.

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“…Concentration of a-CGTase is a costly and time-consuming process. The cost and time requirements are problematic for any catalysis process that requires production of high-quality a-CD [33]. Therefore, enhancing the yields of a-CGTase is of interest.…”

Section: Introductionmentioning

confidence: 99%

Enhanced production of α-cyclodextrin glycosyltransferase in Escherichia coli by systematic codon usage optimization

Liu

et al. 2012

Journal of Industrial Microbiology and Biotechnology

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Enhancing the production of α-cyclodextrin glycosyltransferase (α-CGTase) is a key aim in α-CGTase industries. Here, the mature α-cgt gene from Paenibacillus macerans JFB05-01 was redesigned with systematic codon optimization to preferentially match codon frequencies of Escherichia coli without altering the amino acid sequence. Following synthesis, codon-optimized α-cgt (coα-cgt) and wild-type α-cgt (wtα-cgt) genes were cloned into pET-20b(+) and expressed in E. coli BL21(DE3). The total protein yield of the synthetic gene was greater than wtα-cgt expression (1,710 mg L⁻¹) by 2,520 mg L⁻¹, with the extracellular enzyme activity being improved to 55.3 U mL⁻¹ in flask fermentation. ΔG values at -3 to +50 of the pelB site of both genes were -19.10 kcal mol⁻¹. Functionally, coα-CGTase was equally as effective as wtα-CGTase in forming α-cyclodextrin (α-CD). These findings suggest that preferred codon usage is advantageous for translational efficiency to increase protein expression. Finally, batch fermentation was applied, and the extracellular coα-CGTase enzyme activity was 326 % that of wtα-CGTase. The results suggest that codon optimization is a reasonable strategy to improve the yield of α-CGTase for industrial application.

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“…Also, CGTases can transfer the glycosyl residues to a water molecule (hydrolysis) or a second sugar molecule (disproportionation). These enzymes have proved to be efficient in the synthesis of new glycosyl derivatives of AGs, having more than two glucose units (Svensson et al, 2009a(Svensson et al, , 2009bMathew & Adlercreutz, 2012;Zehentgruber et al, 2011).…”

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confidence: 99%

Synthesis of novel oligomeric anionic alkyl glycosides using laccase/TEMPO oxidation and cyclodextrin glucanotransferase (CGTase)‐catalyzed transglycosylation

Ngo

Linares-Pastén

Grey

et al. 2021

Biotech & Bioengineering

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Modification of alkyl glycosides, to alter their properties and widen the scope of potential applications, is of considerable interest. Here, we report the synthesis of new anionic alkyl glycosides with long carbohydrate chains, using two different approaches: laccase/2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) oxidation of a long-carbohydrate-chain alkyl glycoside and cyclodextrin glucanotransferase (CGTase)-catalyzed elongation of anionic alkyl glycosides. The laccase/TEMPO oxidation of dodecyl β-D-maltooctaoside proceeded efficiently with the formation of aldehyde and acid products. However, depolymerization occurred to a large extent, limiting the product yield and purity. On the other hand, CGTase-catalyzed coupling/disproportionation reactions with α-cyclodextrin and dodecyl β-D-maltoside diuronic acid (DDM-2COOH) or octyl β-D-glucuronic acid (OG-COOH) as substrates gave high conversions, especially when the CGTase Toruzyme was used. It was found that pH had a strong influence on both the enzyme activity and the acceptor specificity. With non-ionic substrates (dodecyl β-D-maltoside and octyl β-Dglucoside), Toruzyme exhibited high catalytic activity at pH 5-6, but for the acidic substrates (DDM-2COOH and OG-COOH) the activity was highest at pH 4. This is most likely due to the enzyme favoring the protonated forms of DDM-2COOH and OG-COOH, which exist at lower pH (pKa about 3).

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Substrate complexation and aggregation influence the cyclodextrin glycosyltransferase (CGTase) catalyzed synthesis of alkyl glycosides

Cited by 9 publications

References 13 publications

Hydrophobic Complexation Promotes Enzymatic Surfactant Synthesis from Alkyl Glucoside/Cyclodextrin Mixtures

Hydrophobic Complexation Promotes Enzymatic Surfactant Synthesis from Alkyl Glucoside/Cyclodextrin Mixtures

Enhanced production of α-cyclodextrin glycosyltransferase in Escherichia coli by systematic codon usage optimization

Synthesis of novel oligomeric anionic alkyl glycosides using laccase/TEMPO oxidation and cyclodextrin glucanotransferase (CGTase)‐catalyzed transglycosylation

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