Improved xylanase Characteristics upon enzyme entrapment Isolated from Thermomyces lanuginosus C9

Xylanases from microbial sources assume basic jobs in an assortment of industrial applications as a biocatalyst, and its applications generally require immobilization on supports to upgrade their stability. Enzyme immobilization is a thrilling decision to show signs of improved strength of enzymatic procedures. In this work, two sorts of polymeric backings (agar-agar and calcium alginate) are utilized to immobilize β-1,4-xylanase from Thermomyces lanuginosus C9 by entrapment, and afterward, biochemical properties of the entangled enzymes were performed. To create immobilized catalyst beads centralization of 4% agar while mix of sodium alginate 5% and calcium chloride 0.4 M was seen as ideal. Ideal reaction time for agar and calcium alginate immobilized protein increments from 10 to 25 and 30 min, separately. The incubation temperature expanded from 70°C to 75°C for agar however stayed unaltered for calcium alginate. The pH profile of free and immobilized xylanase was generally equal in both cases. Be that as it may, both the strategies changed the active boundaries of immobilized β-1,4-xylanase rather than free protein. High sub-atomic load of xylan limits dispersion which brings down the Vmax estimation of immobilized protein while Km value expanded. In contrast with agar-agar, protein immobilized inside calcium alginate display wide thermal stability and kept up 86.6% of its underlying activity at 80°C up to 150 min. Be that as it may, biotechnological portrayal demonstrated that the catalyst reusability was the most surprising discovery, predominantly of agar-agar immobilized xylanase, which held 31% activity after 7 cycles. These outcomes prove the biotechnical and monetary advantages of immobilization which help in an assortment of industrial applications.

Thermo-responsive super porous p(NIPAM) cryogels affords enhanced thermal stability and activity for ɑ-Glucosidase enzyme by entrapping in situ

The concept of using a thermo-responsive p(NIPAM) polymer matrix for enzyme immobilization with lower critical solution temperature (LCST) value is rationalized by availability of the compartmental milieu to enzymes to operate within super porous 3-D matrix with special environmental conditions. Therefore, the enzyme immobilization within a support material will be carried out under the storage conditions of enzymes, generally ~-20 oC to afford unnecessarily loss of enzyme functionality in comparison to the other enzyme entrapment methods. Thus, here ɑ-Glucosidase as a model enzyme was entrapped within thermo-responsive super porous p(NIPAM) cryogels (ɑ-Glu@p(NIPAM) during the synthesis that uses cryogenic condition, ~-20 oC. The LSCT value for the prepared p(NIPAM) based cryogels were determined as 34.6±1.2 oC. The immobilization yield, immobilization efficiency, and activity recovery% values were calculated as 89.4±3.1, 66.2±3.3, and 74.0±3.3%, respectively at pH 6.8 and 37 oC for ɑ-Glu@p(NIPAM) cryogel system. Interestingly, the optimum working conditions were achieved as 25 oC and pH 6.8 with higher activity, 98.4±0.2% for the prepared ɑ-Glu@p(NIPAM) cryogel system. The operational and storage stability studies revealed that the prepared ɑ-Glu@p(NIPAM) cryogel system possessed much better operational and storage stability than free ɑ-Glu enzyme e.g., more than 50% activity after 10th usage and 10-day room temperature storage time. Moreover, the kinetic parameters such as Km and Vmax of free-Glu enzyme and ɑ-Glu@p(NIPAM) cryogel system were calculated by non-linear Michaelis-Menten equation.

Enzyme Entrapment, Biocatalyst Immobilization without Covalent Attachment

In entrapment an active species, which is often a catalyst, is trapped within a material by a solid or gel forming event; thus, it becomes dispersed within the solid or...

Comparative Study on Enzyme Immobilization Using Natural Hydrogel Matrices—Experimental Studies Supported by Molecular Models Analysis

Currently, great attention is focused on conducting manufacture processes using clean and eco-friendly technologies. This research trend also relates to the production of immobilized biocatalysts of industrial importance using matrices and methods that fulfill specified operational and environmental requirements. For that reason, hydrogels of natural origin and the entrapment method become increasingly popular in terms of enzyme immobilization. The presented work is the comparative research on invertase immobilization using two natural hydrogel matrices—alginate and gelatin. During the study, we provided the molecular insight into the structural characteristics of both materials regarding their applicability as effective enzyme carriers. In order to confirm our predictions of using these hydrogels for invertase immobilization, we performed the typical experimental studies. In this case, the appropriate conditions of enzyme entrapment were selected for both types of carrier. Next, the characterization of received invertase preparations was made. As a final experimental result, the gelatin-based hydrogel was selected as an effective carrier for invertase immobilization. Hereby, using mild conditions and a pro-ecological, biodegradable matrix, it was possible to obtain very stable and reactive biocatalyst. The choice of gelatin-immobilized invertase preparation was compatible with our predictions based on the molecular models of hydrogel matrices and enzyme used.

Sucrose-mediated heat-stiffening microemulsion-based gel for enzyme entrapment and catalysis

Formation of a thermo-stiffening microemulsion-based-gel showing the nanoconfinement effect of carbohydrates as an efficient batch bioreactor for entrapped enzymes has been reported.

More than a Confinement: “Soft” and “Hard” Enzyme Entrapment Modulates Biological Catalyst Function

Catalysis makes chemical and biochemical reactions kinetically accessible. From a technological point of view, organic, inorganic, and biochemical catalysis is relevant for several applications, from industrial synthesis to biomedical, material, and food sciences. A heterogeneous catalyst, i.e., a catalyst confined in a different phase with respect to the reagents’ phase, requires either its physical confinement in an immobilization matrix or its physical adsorption on a surface. In this review, we will focus on the immobilization of biological catalysts, i.e., enzymes, by comparing hard and soft immobilization matrices and their effect on the modulation of the catalysts’ function. Indeed, unlike smaller molecules, the catalytic activity of protein catalysts depends on their structure, conformation, local environment, and dynamics, properties that can be strongly affected by the immobilization matrices, which, therefore, not only provide physical confinement, but also modulate catalysis.

Enzyme Entrapment in Amphiphilic Myristyl-Phenylalanine Hydrogels

Supramolecular amino acid and peptide hydrogels are functional materials with a wide range of applications, however, their ability to serve as matrices for enzyme entrapment have been rarely explored. Two amino acid conjugates were synthesized and explored for hydrogel formation. These hydrogels were characterized in terms of strength and morphology, and their ability to entrap enzymes while keeping them active and reusable was explored. It was found that the hydrogels were able to successfully entrap two common and significant enzymes—horseradish peroxidase and -amylase—thus keeping them active and stable, along with inducing recycling capabilities, which has potential to further advance the industrial biotransformation field.