Directed immobilization of CGTase: The effect of the enzyme orientation on the enzyme activity and its use in packed-bed reactor for continuous production of cyclodextrins

Schöffer, Jéssie da Natividade; Matte, Carla Roberta; Charqueiro, Douglas Santana; Menezes, Eliana Weber de; Costa, Tânia Maria Haas; Benvenutti, Edílson V.; Rodrigues, Rafael C.; Hertz, Plinho Francisco

doi:10.1016/j.procbio.2017.04.041

Cited by 26 publications

(13 citation statements)

References 63 publications

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“…The immobilization of Leloir GTs enables a simplification of the reactor’s structure and allows for precise control of the enzymatic glycosylation process [255,256,257]. The immobilization of glycosyltransferases has been achieved by attachment onto solid supports [78,128,258,259,260], entrapment inside a porous carrier [252,261], or cross-linking in larger aggregates (CLEA) [262], as is shown in Figure 14. Furthermore, after immobilization the reusability, thermal, pH, and operational stability of the enzymes was often increased [128,262,263].…”

Section: Reactor Engineering For (Non)-leloir Glycosyltransferasesmentioning

confidence: 99%

“…A packed-bed reactor is a commonly used system for continuous production with a heterogeneous biocatalyst, especially because one can immobilize simultaneously different enzymes. Schöffer et al used glutaraldehyde-activated chitosan spheres [259] and amino- or thiol- functionalized silica particles [260] as a support for cyclodextrin glucosyltransferase immobilization. The silica-based biocatalyst was successfully applied in a packed-bed reactor for continuous cyclodextrin production and maintained 100% of its initial activity after 200 h, whereas activity of the chitosan-based catalyst decreased to 78% of its initial value already after 50 h. This was ascribed to the super packing of spheres, resulting in the reduction of bed height by 45%, and thus in a decrease in the residence/reaction time.…”

Section: Reactor Engineering For (Non)-leloir Glycosyltransferasesmentioning

confidence: 99%

See 1 more Smart Citation

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Mestrom

Przypis

Kowalczykiewicz

et al. 2019

IJMS

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Enzymes are nature's catalyst of choice for the highly selective and efficient coupling of carbohydrates. Enzymatic sugar coupling is a competitive technology for industrial glycosylation reactions, since chemical synthetic routes require extensive use of laborious protection group manipulations and often lack regio-and stereoselectivity. The application of Leloir glycosyltransferases has received considerable attention in recent years and offers excellent control over the reactivity and selectivity of glycosylation reactions with unprotected carbohydrates, paving the way for previously inaccessible synthetic routes. The development of nucleotide recycling cascades has allowed for the efficient production and reuse of nucleotide sugar donors in robust one-pot multi-enzyme glycosylation cascades. In this way, large glycans and glycoconjugates with complex stereochemistry can be constructed. With recent advances, LeLoir glycosyltransferases are close to being applied industrially in multi-enzyme, programmable cascade glycosylations.hydroxynitrile lyases catalyzed the enzymatic hydrolysis of the glycoside amygdalin [3]. Moving almost two centuries forward, the largest volumetric biocatalytic industrial process is the application of glucose isomerase for the production of high fructose syrup for food and drink applications, producing fructose from glucose at 10 7 tons per year [4]. The secret of the success of enzymes in the production or treatment of carbohydrates and glycosides is their exquisite stereo-and regioselectivity. The excellent selectivity of enzymes is required due to the diversity of structural features of carbohydrates [5], comprising d-and l-epimers, ring size, anomeric configuration, linkages, branching, and oxidation state(s). Since drug targets often exhibit specificity for all of these structural features, the production process should not contain any side-products to prevent undesired side-effects [6].The challenge in the synthesis of carbohydrates is their wide variety of functionalities and stereochemistry ( Figure 1). (Poly)hydroxyaldehydes containing a terminal aldehyde are referred to as aldoses and (poly)hydroxyketones are defined as ketoses. In aqueous solutions, monosaccharides form equilibrium mixtures of linear open-chain and ring-closed 5-or 6-membered furanoses or pyranoses, respectively. For aldoses, the asymmetric ring forms at C-1. For ketoses, it closes at C-2 as an axial (α) or equatorial (β) hemiacetal or hemiketal, respectively (commonly defined as the anomeric center). A glycosidic linkage is a covalent O-, S-, N-, or C-bond connecting a monosaccharide to another residue resulting in a glycoside, while glucoside is specific for a glucose moiety. The equatorial or axial position of the glycosidic bond is referred to as α-(axial) or β-linkage (equatorial). The number of carbohydrates linked via glycosidic bonds can be subdivided into oligosaccharides with two to ten linked carbohydrates, while polysaccharides (glycans) contain more than ten glycosidic bonds. A glycan either con...

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Section: Reactor Engineering For (Non)-leloir Glycosyltransferasesmentioning

confidence: 99%

Section: Reactor Engineering For (Non)-leloir Glycosyltransferasesmentioning

confidence: 99%

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Mestrom

Przypis

Kowalczykiewicz

et al. 2019

IJMS

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“…As for synthetic catalysts, many advantages arise from a proper enzyme immobilization such as improved operational activity, stability, reusability, selectivity, specificity (critical in resolutions of racemic mixtures), purification of enzymes or resistance to deleterious compounds [ [8] , [9] , [10] , [11] , [12] , [13] , [14] ]. In the absence of a specific design immobilization of proteins often leads to uncontrolled enzymes orientations which can alter the resulting activity probably due to distortion of the enzyme conformation, or a limited access to the active site and difficulty of mass transfer [ [15] , [16] , [17] ]. Orientation depends greatly on the nature of biocatalyst, the properties of the solid support and the method used for immobilization [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

“…Addressing enzyme orientation, understanding enzyme-surface interaction and test it experimentally still remain challenging tasks especially in confined environment [ [21] , [22] , [23] , [24] ]. A control of enzyme orientation is increasingly important for several purposes such as methods development [ 25 , 26 ], biosensors and biochips [ 7 ], catalysis [ 15 , 27 ], electrocatalysis [ 21 , [28] , [29] , [30] , [31] , [32] ]. Amongst these developments examples of oriented enzyme immobilization in a confined environment are available thus stating the great interest of this control [ 12 , 15 , 16 , 19 , 33 ].…”

Section: Introductionmentioning

confidence: 99%

Site directed confinement of laccases in a porous scaffold towards robustness and selectivity

Yang

Backov

Blin

et al. 2021

Biotechnology Reports

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“…Moreover, biofuel production from trans-esterification of natural oils and production of enantiomeric compounds using enzymes are applications with great industrial and pharmaceutical interest [ 6 , 7 , 8 , 9 , 10 , 11 ]. Recent reports in the literature have shown that enzyme immobilization strategies may control the efficiency of the biocatalyst toward improved properties when compared with the behavior of non-immobilized enzyme [ 12 , 13 , 14 , 15 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

Monitoring the Activity of Immobilized Lipase with Quinizarin Diester Fluoro-Chromogenic Probe

et al. 2017

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Quinizarin diester is used as a fluoro-chromogenic substrate of the activity of lipase supported in poly(methylmetacrylate) beads (CALB, Novozym® 435) dispersed in organic solvents. The monoester and diester of quinizarin are both non-fluorescent species contrasting with the enzymatic product quinizarin that shows optical absorption in the visible region and strong fluorescence signal. The enzymatic conversion is accomplished by spectroscopic measurements and it follows a sigmoid curve from which the mean reaction time of the enzymatic process can be determined. This parameter indicates the enzyme activity of the immobilized lipase. Its dependency with the amount of lipase allowed the determination of the ratio of the catalytic rate and the Michaelis constant (kc/Km) and the experimental value found was (1.0 ± 0.1) × 10−2 mg−1/min in the case of quinizarin diacetate.

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Directed immobilization of CGTase: The effect of the enzyme orientation on the enzyme activity and its use in packed-bed reactor for continuous production of cyclodextrins

Cited by 26 publications

References 63 publications

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Site directed confinement of laccases in a porous scaffold towards robustness and selectivity

Monitoring the Activity of Immobilized Lipase with Quinizarin Diester Fluoro-Chromogenic Probe

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