Increasing the thermostability of sucrose phosphorylase by multipoint covalent immobilization

Cerdobbel, An; Desmet, Tom; Winter, Karel De; Maertens, Jo; Soetaert, Wim

doi:10.1016/j.jbiotec.2010.07.029

Cited by 44 publications

(28 citation statements)

References 33 publications

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“…[34] Moreover, the pH range wasf ound to have been broadened (Supporting Information, Figure S4), indicating enhanced operational stability.T he latter was further investigated by determining the stability of the CP CLEAs at 50 8 8C ( Figure 2). Immobilization boosted the half-life at 50 8 8Cf rom 34 ht oa lmost1 1d ays.Asimilar pattern was observed upona ddition of the IL AMMOENG TM 101 or EtOAc, revealing close correlation between thermal and solvent stability.…”

Section: Exploring the Glycosylation Potential Of Cpmentioning

confidence: 99%

Chemoenzymatic Synthesis of β‐D‐Glucosides using Cellobiose Phosphorylase from Clostridium thermocellum

et al. 2015

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Over the past decade,d isaccharide phosphorylases have been successfully applied for the synthesiso fn umerous a-glucosides. In contrast, much less researchh as been done with respect to the production of b-glucosides. Although cellobiose phosphorylase was alreadys uccessfully used for the synthesiso fv arious disaccharides and branched trisaccharides,i ts glycosylation potentialt owards small organic compounds has not been explored to date. Unfortunately,d isaccharide phosphorylases typically have av ery low affinity for non-carbohydrate acceptors,w hich urges the addition of solvents.T he ionic liquid AMMOENG TM 101 ande thyl acetate were identifieda st he most promising solvents,a llowing the synthesis of various b-glucosides.N extt oh exyl, heptyl,o ctyl,n onyl, decyl and undecyl b-d-glucopyranosides, also the formation of vanillyl 4-O-b-d-glucopyranoside,2 -phenylethyl b-d-glucopyranoside, bcitronellyl b-d-glucopyranoside and 1-O-b-d-glucopyranosyl hydroquinone was confirmed by nuclear magnetic resonance spectroscopy and mass spectrometry.M oreover, the stability of cellobiose phosphorylase could be drastically improvedb yc reating cross-linked enzyme aggregates,w hilet he efficiency of the biocatalyst for the synthesis of octyl b-d-glucopyranoside wasd oubled by imprinting with octanol. Theu sefulness of the latter system was illustrated by performing three consecutive batch conversions with octanoli mprinted cross-linked enzyme aggregates, yieldingr oughly 2gof octyl b-d-glucopyranoside.

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Section: Exploring the Glycosylation Potential Of Cpmentioning

confidence: 99%

Chemoenzymatic Synthesis of β‐D‐Glucosides using Cellobiose Phosphorylase from Clostridium thermocellum

et al. 2015

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“…EC‐HFA and IB‐350 also included an ethylenediamine arm that binds the support surface to the epoxide moieties that give multipoint binds with the proteins and avoids hydrophobic protein–particle interactions . Our enzymes are not affected by the arm in the IB series, but when the binding was in EC‐HFA, D,L‐hydantoinase and N ‐carbamoyl‐amino acid racemase were not active, probably due to steric hindrances.…”

Section: Resultsmentioning

confidence: 97%

Immobilization of a multi‐enzyme system for L‐amino acids production

Rodríguez-Alonso

Rodríguez‐Vico

Heras‐Vázquez

et al. 2015

J of Chemical Tech & Biotech

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BACKGROUND Four enzymes were immobilized for the production of optically pure natural and non‐natural L‐amino acids via the ‘Double‐Racemase Hydantoinase Process’. Immobilization constitutes an empirical process, and for each enzyme we tested 11 carriers with different functional groups; epoxide, EC‐EP, EC‐HFA, IB‐150 and IB‐350, carboxylic acid IB‐C435, quaternary ammonium IB‐A161, IB‐A171 and IB‐A369, aromatic group IB‐S861, and hydroxyl group IB‐S60S and IB‐S60P. RESULTS Each protein showed preference for binding on one or several supports: D,L‐hydantoinase/IB‐350, hydantoin racemase/EC‐EP, L‐carbamoylase/EC‐EP and carbamoyl racemase/IB‐A161. The process was optimized for each enzyme by modifying temperature, pH and ionic strength. For the enzymatic cascade, it was demonstrated that it was essential to use supports having homogeneous characteristics. The product of the first reaction is the substrate of the next one, and so on. Free mass diffusion from one enzyme to the other is crucial to avoid retention on the support and to maintain the protein–substrate interaction constant. CONCLUSION It was proved that support size, weight and hydrophobicity must be homogeneous to avoid the formation of separate layers of the matrix selected. For this reason, the support IB‐350 was used for the four enzymes, even though it may not be the most efficient one for some of them. © 2015 Society of Chemical Industry

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“…Similar results have been often found after immobilization of enzymes. [29][30][31][32][33] This result might be due to multi-point ionic interaction between the enzyme and the Sepharose matrix containing cationic Ni 2+ ions. In general, the activity of the immobilized enzyme is more resistant than that of the soluble form against heat and denaturing agents.…”

Section: )mentioning

confidence: 99%

Characterization and immobilization on nickel-chelated Sepharose of a glutamate decarboxylase A from Lactobacillus brevis BH2 and its application for production of GABA

Lee

Jeon

2014

Bioscience, Biotechnology, and Biochemistry

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A gene encoding glutamate decarboxylase A (GadA) from Lactobacillus brevis BH2 was expressed in a His-tagged form in Escherichia coli cells, and recombinant protein exists as a homodimer consisting of identical subunits of 53 kDa. GadA was absolutely dependent on the ammonium sulfate concentration for catalytic activity and secondary structure formation. GadA was immobilized on the metal affinity resin with an immobilization yield of 95.8%. The pH optima of the immobilized enzyme were identical with those of the free enzyme. However, the optimum temperature for immobilized enzyme was 5 °C higher than that for the free enzyme. The immobilized GadA retained its relative activity of 41% after 30 reuses of reaction within 30 days and exhibited a half-life of 19 cycles within 19 days. A packed-bed bioreactor with immobilized GadA showed a maximum yield of 97.8% GABA from 50 mM l-glutamate in a flow-through system under conditions of pH 4.0 and 55 °C.

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Increasing the thermostability of sucrose phosphorylase by multipoint covalent immobilization

Cited by 44 publications

References 33 publications

Chemoenzymatic Synthesis of β‐D‐Glucosides using Cellobiose Phosphorylase from Clostridium thermocellum

Chemoenzymatic Synthesis of β‐D‐Glucosides using Cellobiose Phosphorylase from Clostridium thermocellum

Immobilization of a multi‐enzyme system for L‐amino acids production

Characterization and immobilization on nickel-chelated Sepharose of a glutamate decarboxylase A from Lactobacillus brevis BH2 and its application for production of GABA

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