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•Direct conversion of cellulose to gluconic acid was achieved on immobilized enzyme.•Tri-enzyme biocatalyst of cellulase, glucose oxidase and catalase was prepared.•Enzymes were ...covalently immobilized on reversibly soluble polymers carrier.•The gluconic acid was obtained from cellulose with conversion rate of 61.41%.
The difficulties in the process of cellulose cascade conversion based on immobilization technology lies in the recycling enzymes from rich solid-containing straw hydrolysate and the incompatibility of conventional immobilization with this process. In this study, three types of enzyme (cellulase, glucose oxidase and catalase) were successfully immobilized on a reversible soluble Eudragit L-100. Through the determination of the preparation conditions, enzymatic properties and catalytic conditions, the co-immobilized enzyme was applied to the catalytic reaction of one-pot conversion of corn straw to gluconic acid. The yield of gluconic acid achieved 0.28 mg/mg, conversion rate of cellulose in corn straw to gluconic acid reached 61.41%. The recovery of co-immobilized enzyme from solid substrate was achieved by using reversible and soluble characteristics of the carrier. After 6 times of recycling, the activity of co-immobilized enzyme was maintained at 52.38%, confirming the feasibility of multi-enzyme immobilization strategy using reversible soluble carrier in cascade reactions.
The synergistic combination of current biotechnological and nanotechnological research has turned to multienzyme co-immobilization as a promising concept to design biocatalysis engineering. It has ...also intensified the development and deployment of multipurpose biocatalysts, for instance, multienzyme co-immobilized constructs, via biocatalysis/protein engineering to scale-up and fulfil the ever-increasing industrial demands. Considering the characteristic features of both the loaded multienzymes and nanostructure carriers, i.e., selectivity, specificity, stability, resistivity, induce activity, reaction efficacy, multi-usability, high catalytic turnover, optimal yield, ease in recovery, and cost-effectiveness, multienzyme-based green biocatalysts have become a powerful norm in biocatalysis/protein engineering sectors. In this context, the current state-of-the-art in enzyme engineering with a synergistic combination of nanotechnology, at large, and nanomaterials, in particular, are significantly contributing and providing robust tools to engineer and/or tailor enzymes to fulfil the growing catalytic and contemporary industrial needs. Considering the above critics and unique structural, physicochemical, and functional attributes, herein, we spotlight important aspects spanning across prospective nano-carriers for multienzyme co-immobilization. Further, this work comprehensively discuss the current advances in deploying multienzyme-based cascade reactions in numerous sectors, including environmental remediation and protection, drug delivery systems (DDS), biofuel cells development and energy production, bio-electroanalytical devices (biosensors), therapeutical, nutraceutical, cosmeceutical, and pharmaceutical oriented applications. In conclusion, the continuous developments in nano-assembling the multienzyme loaded co-immobilized nanostructure carriers would be a unique way that could act as a core of modern biotechnological research.
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•Immobilized multienzymatic systems represent a versatile platform for biocatalysis.•Prospective nano-carriers as host for immobilized multienzymatic systems are reviewed.•Biotechnological applications of immobilized multienzymatic systems are outlined.•Improvement of stability and reusability of the co-immobilized enzymes is emphasized.•Future trends for widespread applications of multienzymatic systems are highlighted.
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•The trends in enzymes-conjugated materials as robust catalytic cues are given.•Smart chemistry interplays a critical role between enzyme and multifunctional matrices.•Industrial ...sector requires momentous efforts to revolutionize modern biocatalysis.•The deployment of a double-enzymatic system, regardless of support materials, in applied settings, is remarkable.
Enzymes have received extensive research due to their high selectivity, efficiency, and environmental protection. However, the lack of stability and reusability drastically hinders the advancement of various enzyme-mediated applications. To assure functionality and the potential for enzyme recycling after use, the immobilization of enzymes has been significantly demanded in a broad range of industrial applications. Nano biocatalyst is a recently developed catalyst that synergistically combines biotechnology and modern nanotechnology. It provides attractive possibilities for enhancing enzyme activity, stability, capability, and engineering performances in biotechnological processes. Due to their large interfacial area, greater activity, and long-term sustainability, nanostructured materials have been shown to be effective substrates for the immobilization of enzymes. In order to increase enzymatic activity as a biocatalyst in various bio-medical fields, food and chemical industries, as well as in bio-sensing and degradation of environmental pollutants, different carbon-based nanostructures coated with various polymeric materials have been studied as carriers for enzyme immobilization. Co-immobilization, which relates to the concurrent immobilization of two or more enzymes, has also been highlighted. The significance of this study can aid in enhancing the efficacy, viability, and economics of numerous industrial processes that rely on enzymatic reactions. This article highlights the various approaches and purposes established in this domain and the quick progress made to design isolation technologies that are more effective, durable, and long-lasting for a wide range of applications & enzymes. Diffusion limitations, leaching, cost, loss of activity and scalability are some problems related to this technique. This review provides a comprehensive point of view on enzyme immobilization that compresses a critical analysis of immobilization techniques and carriers, biocatalytic matrices, the influence of vital carrier characteristics on biocatalyst performance, trends toward nanotechnology, and in-depth case studies that are reflective of biocatalysis trying to promote sustainable development. Finally, the challenges in using nanostructures to immobilize enzymes and their potential in the future have been discussed.
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•Chitin/chitosan composite microspheres (CCMs) were designed with controllable DDA values.•The one-step purified and immobilization of protein was achieved using CCMs.•The CCMs ...realize the efficiently co-immobilization of enzyme and PLP.•The polyethyleneimine modified CCMs exhibited strong affinity of PLP.•The designed biocatalysts provided a platform technology for continuous catalysis without addition of PLP.
The concomitant recycling of enzymes and co-factors is a big challenge in biocatalysis. In this study, co-immobilization of the enzyme and the co-factor pyridoxal 5′-phosphate was achieved using chitin-based microspheres as the support. Chitin/chitosan composite microspheres were prepared and then modified with polyethyleneimine. Three pyridoxal 5′-phosphate-independent enzymes were used to demonstrate that the self-sufficient biocatalysts based on chitin/chitosan composite microspheres and polyethyleneimine modified microspheres displayed good generalizability for high recycling efficiency of both enzyme and pyridoxal 5′-phosphate. Furthermore, the self-sufficient biocatalysts were applied in the continuous flow production of cadaverine and maintained good continuous catalysis without the exogenous addition of pyridoxal 5′-phosphate, suggesting that this co-immobilization platform shows promise for application in industrial biocatalysis.
ZIF-67 derived catalysts for Fischer-Tropsch synthesis have attracted much attention in recent years, while there is still a potential to improve their activity and selectivity. In this work, we ...prepared Si/Al co-immobilized Co3O4@SixAly catalysts by in-situ doping tetraethylorthosilicate and aluminum nitrate during the synthesis of ZIF-67. The effects of different Si/Al ratios on the metal-support interaction, acidity and FTS performance were explored. Results indicated that the Co3O4@Si0Al4 catalyst exhibited the best FTS performance with the CO conversion as high as 79.9% and CTY (cobalt time yield) value of 19.5 × 10−5 molCO·gCo−1·s−1, which was ascribed to the moderate metal-support interaction and the most active Co sites. Meanwhile, the Co3O4@Si3Al1 and Co3O4@Si2Al2 catalysts exhibited higher iso-paraffin and olefin selectivity due to more acidic sites.
The Co3O4@Si0Al4 catalyst exhibited the highest FTS activity due to the moderate metal-support interaction (MSI), while the Co3O4@Si3Al1 and Co3O4@Si2Al2 catalysts showed certain hydrocracking and isomerization ability due to the formation of acidic sites. Display omitted
•Co3O4@SixAly catalysts were prepared by in-situ doping method based on ZIF-67.•The Co3O4@Si0Al4 exhibited the highest Fischer-Tropsch synthesis (FTS) activity.•The high activity was due to moderate metal-support interaction and most active sites.•Acidic sites promoted the hydrocracking and isomerization of long chain hydrocarbons.
Drinking water safety cannot be overemphasized. Filamentous fungi have many excellent features for metal removal. Both graphene oxide (GO) and activated carbon (AC) are conventional metal adsorbents, ...but they are not suitable for large-scale use due to high cost. In this study, a low dosage of conidia (2.0 × 104 conidia/mL) of metal-resistant/adapted filamentous fungus Penicillium janthinillum strain GXCR were co-immobilized with a low dosage of 0.5 mg/L GO or 0.5 mg/L AC by embedding in 2% polyvinyl alcohol (PVA)-3% sodium alginate (SA), generating six types of microbead adsorbents (MBAs) to remove metals from a low concentration of either single metal (100 mg/L) or mixed metals (100 mg/L each) of Pb (II), Fe (III) and Cu (II) in drinking water. Fungus GXCR-containing MBAs had higher specific surface areas (SSAs), better mesoporous structures, and a higher removal rate (85–98.99%) of single or mixed metals. Singl-metal adsorptions of MBAs were almost unaffected by temperature changes. MBAs showed a stable removal rate of 87–94% during four cycles of adsorption-desorption of single metal. Single-metal adsorptions were well described by multiple models of Freundlich isotherm with constant values of 0.21–0.432, Langmuir isotherm with constant values of 0.037–0.17, Pseudo-fist-order, Pseudo-second-order, and intra-particle diffusion (IPD). In conclusion, co-immobilization between GXCR, GO and AC can make metal removal more efficient. Adsorption capacity is increased with SSAs but not in the same proportion. Single-metal adsorptions involve multiple mechanisms of monolayer and multilayer adsorptions, external mass transfer, and IPD. IPD is important but not the only one rate-controlling step for single-metal adsorptions.
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•Adsorbents from co-immobilization of fungus, graphene oxide, and activated carbon.•Adsorbents have metal removal rates of 85–98.99% from stimulated drinking water.•Single metal adsorption kinetics follow multiple models and complex mechanisms.•Intra-particle diffusion is not the only one rate-controlling step for adsorption.•Adsorption capacity is increased with surface-areas but not in the same magnitude.
In order to increase the stability of a bi-enzymes system, β-fructofuranosidase and glucose oxidase were co-immobilized by sol-gel encapsulation and its effect on the production of lactosucrose was ...investigated. The as-prepared immobilized bi-enzymes were characterized by Scanning electron microscopy, Fourier transform infrared spectroscopy, Raman spectroscopy and thermogravimetric analysis. The immobilized bi-enzymes retained 85.39% of their initial activities, and showed markedly better pH tolerance than the free enzymes at pH < 6.5. Especially, immobilization significantly (p < 0.01) enhanced the thermal stability of bi-enzymes at high temperatures (≥40 °C), where almost no activity was observed for free enzymes. The immobilized bi-enzymes showed great operational stability, maintaining 82.0% of their initial activities after eight consecutive reuses. Furthermore, the yield of lactosucrose (40.2%) was significantly (p < 0.05) improved by the employ of the immobilized bi-enzymes. These results indicated that the co-immobilization of bi-enzymes was helpful to improve the production of lactosucrose with low costs.
•β-fructofuranosidase and glucose oxidase were successfully co-immobilized.•Immobilized bi-enzymes with good stability was obtained.•By-product glucose was oxidized to gluconic acid by glucose oxidase.•High lactosucrose content was obtained when synthesized by immobilized bi-enzymes.
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•First attempt to realize high-titer production of inositol by co-immobilized multi-enzymes.•An inositol titer of up to 210 g/L was produced.•Prolonged enzyme lifetime decreased ...biomanufacturing cost.•Scaleable multienzyme immobilization for potential industrial application was proposed.
Myo-inositol is widely used in food, feed, pharmaceutical and cosmetic industries, but its great potentials are restricted from its limited supplies and high prices. Inositol is the first biobased product produced by the in vitro biomanufacturing platform containing multi-enzymes in one pot on an industrial scale. However, the in vitro multi-enzyme-based biomanufacturing face the challenge of enzyme deactivation and high cost of the enzymes used. To further prolong the enzyme lifetime and decrease its biomanufacturing cost, here we developed a co-immobilized four-enzyme cocktail using biomimetic mineralized microcapsules composed of polyethyleneimine and silicate for large-scale production. The lumped enzyme half-life of the microcapsules was 55.5 h at 70 °C, which was 5.9 folds of that of the four-enzyme cocktail. An inositol titer of up to 210 g/L was produced, and the cost of enzymes used was decreased by 3.4 times. This is the first attempt to realize high-titer production of inositol by co-immobilized multi-enzymes, laying a foundation of in vitro biomanufacturing based on in vitro synthetic enzyme biosystems.
•Co-immobilized amylase, pectinase and cellulase can be used for juice clarification.•The enzymes covalently attached to the silica gel showed the highest efficiency.•Turbidity change with ...temperature had inverse trend depending on the enzyme amount.
In this study, the use of co-immobilized amylase, pectinase and cellulase enzymes for clarification of apple, grape and pear juices were investigated. Four different co-immobilization method was applied and the highest activity was observed for co-immobilized enzymes onto silica gel. The effect of the enzyme units used in the co-immobilization, reaction temperature and time on turbidity and reducing sugar concentration were optimized by Response Surface Methodology (RSM) via Design Expert Software 8.0.7.1. applying 3-factor Box-Behnken design. Co-immobilized samples were reacted with the substrate solution in batch type and feedback packed bed column reactors and according to turbidity and reducing sugar concentration values, batch reactor efficiency was found to be higher. Finally, freshly squeezed apple, pear and grape juices were interacted with co-immobilized enzymes in the batch reactor system, and turbidity and reducing sugar concentrations were monitored over time. Changes were also observed in antioxidant activities depending on the enzyme treatment.
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