Stability Of Cellulase Research Articles

In-depth recognition of mixed surfactants maintaining the enzymatic activity of cellulases through stabilization of their spatial structures

Mixed surfactants improve the enzymatic hydrolysis of lignocellulosic substrates by enhancing cellulase stability against heat, pH, shear, and air–liquid interface stress. Under conditions of multiple factorial stresses (50 °C, pH 4.8, 180 rpm, and 15.5 cm2 air–liquid interface), cellulase with ternary surfactants (Tween 60/Triton X-114/CTAB, the molar ratio 14:5.5:1) retained 84 % of its activity after 48 h of incubation, representing 1.15 and 1.29 folds that of the cellulase activity with the single Tween 60 and with no surfactants, respectively. This is attributed to the fact that ternary surfactants possess better rheology modulation and air–liquid interface competitiveness. In addition, the computational approach demonstrated that the ternary surfactants were capable of forming stronger hydrophobic and hydrogen-bond interactions with cellulase enzymes, thus maintaining its secondary structure and preventing the detrimental α-helix to β-sheet transformation known to compromise cellulase activity. This synergy offers valuable insights into surfactant-cellulase interactions and supports efficient enzymatic hydrolysis in biorefineries.

Insights of Pichia kudriavzevii SVMS2019 for cellulase production and fermentation into ethanol

The use of lignocellulosic biomass to produce value-added products, such as fuel ethanol, is a current priority for global biobased economy. The quest to explore novel microbes, such as yeasts, to attain sustainable products for an ever-growing population has become highly in demand. This study might be the first to explore the rumen yeast Pichia kudriavzevii SVMS2019 for cellulase production to efficiently utilize wheat straw as biomass, which can further be converted to bioethanol. The optimization process was carried out on pre-treated wheat straw by employing one factor at a time (OFAT) and response surface methodology (RSM). Wheat straw pre-treated using NaOH alkali pretreatment was characterized with FTIR, XRD, and SEM studies. The findings revealed that the P. kudriavzevii SVMS2019 obtained maximum cellulase activity of 44.064 IU/ml at 37.5 °C, pH 3.5, 120 h, and 1.5% of NaOH pre-treated wheat straw (substrate concentration) through RSM. It exhibited 1.25-fold increased production compared to OFAT. The stability of crude cellulase was observed at 45 °C and pH 6. Further, the hydrolyzed reducing sugars produced under optimal conditions opted for fermentation, with a maximum bioethanol content of 12.13 g/l obtained at 37.5 °C in a 24 h interval.

Coupling chemical modification and immobilization of cellulase improves thermal stability and low-melting mixture solvent tolerance for in situ saccharification of bagasse

In situ saccharification of lignocellulosic biomass in low-melting mixture solvents (LoMMSs) is a promising approach for biorefinery. In this study, chemical modification of cellulase coupled with immobilization was designed to improve thermal stability and LoMMS tolerance of cellulase for in situ saccharification. Cellulase was firstly chemically modified with succinic anhydride and ethylenediamine respectively to form succinylated cellulase (S-Ce) and aminated cellulase (A-Ce), then the modified cellulases were immobilized with amino-functionalized metal-organic frameworks to establish ZIF-8/A immobilized A-Ce and S-Ce. Circular dichroism, fluorescence measurements, X-ray diffraction, fourier transform infrared spectra and scanning electron microscopy were applied to characterize the modified cellulase or immobilization carrier. ZIF-8/A immobilized A-Ce presented desired thermal stability and LoMMS tolerance. Reducing sugar yields via ZIF-8/A immobilized A-Ce during in situ saccharification of bagasse maintained stably till Bet/AA concentration reached 50%, and declined relatively slowly compared with free or other immobilized cellulase when Bet/AA concentration further increased to 90%. In situ saccharification of bagasse via ZIF-8/A immobilized A-Ce in concentrated Bet/AA (50%) can be operated at a wide temperature range (50–60 °C). The corresponding reducing sugar yield with ZIF-8/A immobilized A-Ce was 58% higher than that with free cellulase in 50% Bet/AA at 60 °C.

Coffee Pulp Activated Carbon for Immobilizing Cellulase from Aspergillus niger ICP2: Enhancing Enzyme Stability, Activity, and Its Reusability

Cellulase from Aspergillus niger ICP2 was successfully immobilized on coffee pulp activated carbon using adsorption. Carbon was derived from coffee pulp via controlled carbonization at 200 °C for 2 hours, followed by activation with 3M ZnCl2 for 150 minutes. Reusability analysis exhibited over 50% relative activity retention after four cycles. The immobilized cellulase demonstrated stability at pH 4.0-6.0, retaining over 80% relative activity after 4 hours at 37℃. This approach proves economical for enhancing cellulase stability, activity, and reusability.

Effect of Deep Eutectic Solvents on the Activity and Stability of Cellulases and Pectinases

Effect of Deep Eutectic Solvents on the Activity and Stability of Cellulases and Pectinases

Polydopamine modified cellulose nanocrystals (CNC) for efficient cellulase immobilization towards advanced bamboo fiber flexibility and tissue softness

Herein, a green facile approach to improve the flexibility of unbleached bamboo kraft pulp (UBKP) via an immobilized enzyme technology is proposed. Polydopamine (PDA) acts as versatile modification and coating materials of cellulose nanocrystals (CNC) for assembling versatile bio-carriers (PDA@CNC). Cellulase biomacromolecules are efficiently immobilized on PDA@CNC to form cellulase@PDA@CNC nanocomposites. The relative enzyme activity, temperature/pH tolerance, and storage stability of cellulase were significantly improved after immobilization. The degree of polymerization treated UBKP decreased by 5.42 % (25 U/g pulp) compared to the control sample. The flexibility of treated fibers was 6.61 × 1014/(N·m2), which was 96.93 % higher (25 U/g) compared to the control and 3.88 times higher than that of the blank fibers. Cellulase@PDA@CNC performs excellent accessibility to fiber structure and induces high degree of fibrillation and hydrolysis of UBKP fibers, which contributes high softness of obtained tissue handsheets. The bio-carrier PDA@CNC within paper framework may further enhance tissue tensile strength. This study proposes a practical and environmentally friendly immobilization approach of cellulase@PDA@CNC for improving the hydrolysis efficiency and flexibility of UBKP fibers, which provides the possibility to maintain the strength of tissue paper while improving its softness, thus broadening the high-value application of immobilized enzyme technology in tissue production.

Bionanofabrication of Cupric oxide catalyst from Water hyacinth based carbohydrate and its impact on cellulose deconstructing enzymes production under solid state fermentation

One of the most important properties of cellulolytic enzyme is its ability to convert cellulosic polymer into monomeric fermentable sugars which are carbohydrate by nature can efficiently convert into biofuels. However, higher production costs of these enzymes with moderate activity-based stability are the main obstacles to making cellulase-based applications sustainably viable, and this has necessitated rigorous research for the economical availability of this process. Using water hyacinth (WH) waste leaves as the substrate for cellulase production under solid state fermentation (SSF) while treating the fermentation production medium with CuO (cupric oxide oxide) bionanocatalyst have been examined as ways to make fungal cellulase production economically feasible. Herein, a sustainable green synthesis of CuO bionanocatalyst has been performed by using waste leaves of WH. Through XRD, FT-IR, SEM, and TEM analysis, the prepared CuO bionanocatalyst's physicochemical properties have been evaluated. Furthermore, the effect of CuO bionanocatalyst on the temperature stability of raw cellulases was observed, and its half-life stability was found to be up to 9 h at 65 °C. The results presented in the current investigation may have broad scope for mass trials for various industrial applications, such as cellulosic biomass conversion.

Assessing the compatibility of choline-based deep eutectic solvents for the structural stability and activity of cellulase: Enzyme sustain at high temperature

As a new generation of ‘green solvents’ deep eutectic solvents (DESs) represents a promising alternative to the conventional solvents. Their environmental-benign nature and designer properties promote their utility in biocatalysis. Enzymes are marginally stable when exposed to physical/chemical disturbances. One such enzyme is cellulase which is a propitious catalyst for the depolymerization of cellulose under mild conditions. Therefore, their stability is a prerequisite condition to match demands of biorefineries. To address this issue of low stability, activity and thermal denaturation of cellulase, there is a need to find a sustainable and suitable co-solvent that is biocompatible with enzymes ultimately to facilitate their application in bio-industries. In this regard, we synthesized three choline-based DESs, choline chloride (ChCl)-glycerol, ChCl-ethylene glycol and ChCl-lactic acid and employed them to analyze their suitability for cellulase. The present study systematically evaluates the influence of the mentioned DESs on stability, activity and thermal stability of cellulase with the help of various spectroscopic techniques. The spectroscopic analysis revealed that the structural stability and activity of the enzyme were improved in presence of ChCl-glycerol and ChCl-ethylene glycol. The thermal stability was also very well maintained in both the DESs. Interestingly, the relative activity of cellulase was >80 % even after incubation at 50 °C after 48 h for both the DESs. This activity preservation behaviour was more pronounced for ChCl-ethylene glycol than ChCl-glycerol. Moreover, temperature variations studies also reveal promising results by maintain conformational intactness. On the other side, ChCl-lactic acid showed a deleterious effect on the enzyme both structurally as well as thermally. The dynamic light scattering (DLS) analysis provides more specific information about the negative influence of ChCl-lactic acid towards cellulase native structure. This DES induces unavoidable alterations in the enzyme structure which leads to the unfolding of enzyme, ultimately, destabilizing it. Overall, our results present a physical insight into how the enzyme stability and activity depend on the nature of DES. Also, the findings will help to facilitate the development and application of DESs as biocatalytic process.

A study on critical associations of media components on enhanced cellulase production from wild Trichoderma viride and cellulase immobilization on iron-oxide magnetic nanoparticles

Aim: The current study is a preliminary step towards enhancing the cellulase productivity in wild Trichoderma viride which will enable robust valorization of non-edible lignocellulosic biomass through co-generative enzymatic saccharification, specifically concentrating on influence of individual media components on biomass growth and cellulase productivity. Further, cellulase immobilization on iron-oxide magnetic nanoparticles was also achieved that can increase the shelf life of the enzyme. Methodology: The cellulase production in the wild Trichoderma viride was enhanced using media design and formulation. EDC {1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide} functionalized iron-oxide nanoparticles were chosen to act as carriers for cellulase immobilization. The binding efficiency and relative activity were measured in addition to optimal pH and temperature for cellulase bound iron-oxide nanoparticles. Further, the hydrolysis efficiency of immobilized cellulases was also measured after which it was subjected to consecutive hydrolytic cycles to calculate the recycle rate. Results: A maximum growth rate of 60 PCV (Packed cell volume) and total cellulase activity of 7.4 U ml-1 was obtained on media design and formulation. 82.5% binding efficiency was achieved on EDC functionalized iron-oxide magnetic nanoparticles which showed good stability at 5pH and 500C. There was 44.4% activity loss after 5 consecutive hydrolytic cycles which showed steady decline with increased cycle number and finally at the end of the 10th hydrolytic cycle, 22.2% of total relative activity was retained. Interpretation: Unprecedented total cellulase activity from a wild strain was obtained through media design. The stability of cellulases was further enhanced using iron-oxide magnetic nanoparticle immobilization. Key words: Cellulases, Immobilization, Iron-oxide magnetic nanoparticles, Submerged fermentation, Trichoderma viride

Investigating the Impact of Polymer Length, Attachment Site, and Charge on Enzymatic Activity and Stability of Cellulase.

The thermophilic cellulase Cel5a from Fervidobacterium nodosum (FnCel5a) was conjugated with neutral, cationic, and anionic polymers of increasing molecular weights. The enzymatic activity toward an anionic soluble cellulose derivative, thermal stability, and functional chemical stability of these bioconjugates were investigated. The results suggest that increasing polymer chain length for polymers compatible with the substrate enhances the positive impact of polymer conjugation on enzymatic activity. Activity enhancements of nearly 100% were observed for bioconjugates with N,N-dimethyl acrylamide (DMAm) and N,N-dimethyl acrylamide-2-(N,N-dimethylamino)ethyl methacrylate (DMAm/DMAEMA) due to proposed polymer-substrate compatibility enabled by potential noncovalent interactions. Double conjugation of two functionally distinct polymers to wild-type and mutated FnCel5a using two conjugation methods was achieved. These doubly conjugated bioconjugates exhibited similar thermal stability to the unmodified wild-type enzyme, although enzymatic activity initially gained from conjugation was lost, suggesting that chain length may be a better tool for bioconjugate activity modulation than double conjugation.

Influence of natural deep eutectic solvents on stability and structure of cellulase

To address the issue of low stability and short half-life of cellulase, thermal stability and conformation change of Cellic® CTec2 were studied in various natural deep eutectic solvents (NADESs), with the aim of finding green and suitable solvents to enhance its stability, and thus facilitating utilization of lignocellulosic biomass. Fourteen NADESs (20%, v/v), with betaine or choline chloride (ChCl) as hydrogen bond acceptor (HBA), different polyols, amides or organic acids as hydrogen bond donor (HBD) were examined. When compared with the cellulase incubated in buffer at 50 °C, seven NADESs (20% in buffer, v/v) prolonged half-life time of the enzyme, among which betaine/1,4-butanediol and ChCl/1,4-butanediol (molar ratio 1:2) were the most prominent, with half-life time increased by about 11 and 9 folds, respectively, and >80% residual activity remained after 72 h. The influence of HBDs was more evident than HBAs in the aspect of cellulase stability, particularly HBDs of polyols (esp. diols) were better than amides, whereas HBDs of organic acids seemed to be destructive for cellulase. Associated conformational changes of cellulase during incubation in different solvents were also characterized with fluorescence spectrophotometer and circular dichroism spectrometer. The result demonstrated that cellulase in those NADESs enhancing its stability exhibited a higher fluorescence intensity, as well as minor spectrum intensity decreased over time, when comparing with that in buffer. Further investigation of NADESs concentration suggested that 10–20% (v/v) was optimal for stability enhancement, among which 15% and 20% (v/v) betaine/1,4-butanediol exhibited the best performance, evidenced by relative activity of about 450% after 48 h incubation at 50 °C, showing great promise in enhancing celluase stability.

Sustainable green approach to synthesize Fe3O4/α-Fe2O3 nanocomposite using waste pulp of Syzygium cumini and its application in functional stability of microbial cellulases

Synthesis of nanomaterials following green routes have drawn much attention in recent years due to the low cost, easy and eco-friendly approaches involved therein. Therefore, the current study is focused towards the synthesis of Fe3O4/α-Fe2O3 nanocomposite using waste pulp of Jamun (Syzygium cumini) and iron nitrate as the precursor of iron in an eco-friendly way. The synthesized Fe3O4/α-Fe2O3 nanocomposite has been extensively characterized through numerous techniques to explore the physicochemical properties, including X-ray diffraction, Fourier transform infrared spectroscopy, Raman spectroscopy, Ultraviolet-Vis spectroscopy, field emission scanning electron microscope, high resolution transmission electron microscope and vibrating sample magnetometer. Further, efficiency of the Fe3O4/α-Fe2O3 nanocomposite has been evaluated to improve the incubation temperature, thermal/pH stability of the crude cellulase enzymes obtained from the lab isolate fungal strain Cladosporium cladosporioides NS2 via solid state fermentation. It is found that the presence of 0.5% Fe3O4/α-Fe2O3 nanocomposite showed optimum incubation temperature and thermal stability in the long temperature range of 50–60 °C for 15 h along with improved pH stability in the range of pH 3.5–6.0. The presented study may have potential application in bioconversion of waste biomass at high temperature and broad pH range.

Preparation of Cross-Linked Cellulase Aggregate and Application to Cool Finishing of Cotton Fabric

Cellulase was immobilized by cross-linked enzyme aggregation to improve the stability of cellulase. The prepared cross-linked cellulase aggregates (CLCAs) and ice silicone oil were used for the cool finishing of cotton fabric. The results showed that the CLCAs extended the cellulase stability compared to free cellulase. The surface softness, smoothness, moisture permeability, and air permeability of the cotton fabric increased after CLCAs and ice silicone oil treatment. Shearing rigidity of the treated sample was 0.44 cN/(cm·deg), bending rigidity was 0.0069 cN cm, and the drape coefficient was 29.3%. Coefficient of kinetic friction of the treated sample was 0.186. The capillary effect of the treated fabric was 12.1 cm/(30 min). Air permeability was 354.3 L/(m2·s). Moisture penetrability was 3.912 g/(m2·d). The thermal and water-vapor resistance were 0.0194 m2·°C/W and 4.691 Pa·m2/W, respectively.

Metal-organic frameworks coupling simultaneous saccharication and fermentation for enhanced butyric acid production from rice straw under visible light by Clostridium tyrobutyricum CtΔack::cat1

Here, Metal-Organic Frameworks (MOFs) coupling simultaneous saccharification and fermentation for butyric acid production using rice straw was constructed. Clostridium tyrobutyricum Δack::cat1, with deleted ack gene and overexpressed cat1 gene, was used as the butyric-acid-fermentation strain. MOFs was employed as a photocatalyst to improve butyric acid production, as well as a cytoprotective exoskeleton with immobilized cellulase for the hydrolysis of rice straw. Thus, the survival of MOFs-coated strain, the thermostability and pH stability of cellulase both remarkably increased. As a result, 55% of rice straw was hydrolyzed in 24 h, and the final concentration of butyric acid in visible light was increased by 14.23% and 29.16% compared to uncoated and coated strain without visible light, respectively. Finally, 26.25 g/L of butyric acid with a productivity of 0.41 g/L·h in fed-batch fermentation was obtained. This novel process inspires green approach of abundant low-cost feedstocks utilization for chemical production.

Detergent-compatible fungal cellulases

Detergent enzymes are currently added to all powder and liquid detergents that are manufactured. Cellulases, lipases, amylases, and proteases are used in the detergency to replace toxic phosphates and silicates and to reduce high energy consumption. This makes the use of enzymes in detergent formulation cost effective. Fungi are producers of important extracellular enzymes for industrial use. The fungal and bacterial cellulases maintain the shape and color of the washed garments. There is a high demand for cellulases at the market by detergent industries. With this high demand, genetic engineering has been a solution due to its high production of detergent-compatible cellulases. Fungi are the famous source for detergent-compatible cellulases production, but still, there is a lack of the cost-effective process of alkaline fungal cellulase production. Review papers on detergent-compatible bacterial cellulase and amylase and detergent-compatible fungal and bacterial proteases and lipases are available, but there is no review on detergent fungal cellulases. This review aims to highlight the production, properties, stability, and compatibility of fungal cellulases. It will help other academic and industrial researchers to study, produce, and commercialize the fungal cellulases with good aspects.

Synergistic effect of ionic liquid and surfactant for enzymatic hydrolysis of lignocellulose by Paenibacillus sp. LLZ1 cellulase

Stable cellulases are critical for in situ hydrolysis of ionic liquid-pretreated lignocellulose, and surfactants were reported to improve the stability of cellulase in practical circumstances. In this study, synergistic effect of Tween-80 and [Emim]OAc was first identified to enhance the hydrolysis of lignocellulose by Paenibacillus sp. LLZ1 cellulase, proving the effectiveness of surfactants in in situ hydrolysis. Protein fluorescence and near-UV circular dichroism spectroscopy analyses illustrated that ionic liquids might move local residues and alter global tertiary structure of cellulase, leading to higher instability. The addition of Tween-80 might turn the microenvironment of cellulase residues more hydrophilic and reduce the contact of protein with deactivation factors. The synergistic system was further applied in the in situ hydrolysis of microcrystalline cellulose and bagasse cellulose by Paenibacillus cellulase. The enzymatic hydrolysis rate of microcrystalline cellulose was enhanced by 2.5-fold and the final conversion was improved by 45%. Scanning electron microscopy demonstrated the substrate solubilizing effect during reaction by monitoring the morphological changes of lignocellulosic substrates. This study highlighted the potential of applying synergistic reaction system in in situ enzymatic hydrolysis of lignocellulose, and revealed the inherent mechanism initially.

Effects of Rhamnolipids on Enzymatic Hydrolysis of Bamboo Biomass and Mechanism

The lignin present in lignocellulose seriously affects the efficiency of cellulose enzymatic hydrolysis. In addition, lignin adsorbs high-cost cellulase, causing greater economic losses. Lignin can also disturb the site of action of cellulase and reduce the efficiency of hydrolysis. Therefore, if lignin is removed or surface modified before cellulose enzymatic hydrolysis, the enzymatic hydrolysis efficiency of lignocellulosic biomass will be greatly improved. In this paper, the cellulose enzymatic properties of bamboo biomass being treated with dilute acid and alkaline under the intervention of biosurfactant rhamnolipid were evaluated. The effects of rhamnolipids on the adsorption characterization of cellulose on pretreated bamboo were studied. Besides, the inter-communication between rhamnolipids and cellulose was investigated by fluorescence probe. The results showed that rhamnolipids could have a positive effect on the enzymatic hydrolysis of bamboo biomass by reducing the non-productive adsorption of cellulase on the surface of lignocellulose. The outcome illustrated that cellulase could be combined with rhamnolipids micelles, participating in the formation of rhamnolipids micelles, thereby increasing the internal hydrophobicity of the micelles, but could not change the properties of rhamnolipids micelles higher than one CMC (Critical Micelle Concentration). It can be seen that the interaction between rhamnolipids and cellulase is beneficial to enhance the stability and enzymatic activity of cellulase, thereby improving the enzymatic hydrolysis efficiency of cellulose in biomass. Based on these results, a theoretical knowledge about the mechanism of enhancing the enzymatic hydrolysis efficiency of lignocellulose by biosurfactants rhamnolipids is provided.

Clarification of tangerine juice using cellulases from Pseudoyma sp.

Tangerine juice was treated with crude extract containing cellulase from Pseudozyma sp. obtained by liquid fermentation. The thermal stability of cellulase was investigated by incubating crude extract at different temperatures and times. The pulp, obtained from tangerine, was pasteurized at 85°C for 5min and then used in a clarification process with a Doehlert experimental design. The results showed that the cellulase obtained from Pseudozyma sp. is thermostable at temperatures of 60, 70 and 90°C and retained 98%, 88% and 80% of activity, respectively, after a 1-h incubation time. The optimum conditions for clarification were verified by varying the enzyme extract concentration (%, v v-1) and the time (minutes) in a shaker at 150rpm, at 50°C. The optimum condition for clarification was obtained in the 80th min with a 1.25% enzymatic extract concentration (v v-1), resulting in a reduction of tangerine juice viscosity by 65%. The analysis of physical and chemical parameters of tangerine juice after clarification showed that the enzyme extract improved the process responsible for the clarification of tangerine juice. The results are promising since this is a methodology that can be used in the citrus juice industry.

Involvement of Xyr1 and Are1 for Trichodermapepsin Gene Expression in Response to Cellulose and Galactose in Trichoderma reesei.

The secretome of Trichoderma reesei contains a mixture of cellulases, hemicellulases, amylases, proteases, and lipases that synergistically degrade plant biomass. Trichodermapepsin (TrAsP), the most prominent protease of T. reesei, affects the stability of cellulases. Similar to cellulase production, TrAsP production also depends on carbon and nitrogen sources. Unlike the cellulase mechanism, the regulatory mechanism of TrAsP remains unknown. Therefore, this study aimed to determine the effect of the main cellulase regulator Xyr1 and nitrogen regulator Are1 on trasp regulation. Cellulase inducer Avicel and TrAsP inducer galactose were used as carbon sources. qRT-PCR analysis revealed that Xyr1 and Are1 acted as a repressor and an activator for trasp expression, respectively. Compared to Avicel, relative expression was higher in galactose. The binding motifs of Xyr1 and Are1 were located in upstream of the trasp promoter. From promoter deletant analysis using the β-glucuronidase reporter gene, the area from - 870bp to - 670bp was identified as the only region for positive regulation and there were both binding motifs of Xyr1 and Are1. Reporter assay of mutants confirmed functions of downregulation of Xyr1 and upregulation of Are1. Electrophoretic mobility shift assay demonstrated the binding ability of Xyr1 and Are1 to the particular binding motifs and their functionality was confirmed. Further, this study demonstrated that Cre1, Xpp1, and Pac1 downregulate trasp expression similar to that in cellulase regulation mechanism. These results demonstrate that transcriptional regulators of cellulase control trasp expression and suggest the possibility of the existence of specific protease regulators in T. reesei.

Characterization of magnetic nanoparticle–immobilized cellulases for enzymatic saccharification of rice straw

Cellulases convert lignocellulosic biomass into fermentable sugars which further act as substrate for ethanol production. Our previous research was focused on enzymatic hydrolysis of rice straw by free cellulases for production of fermentable sugars. Immobilization is a powerful tool to increase the stability and reusability of cellulases besides improving the economy of ethanol production process from rice straw. In the present study, cellulase produced from Aspergillus fumigatus was immobilized on magnetic nanoparticles by using glutaraldehyde cross linker with a binding efficiency of 65.55%. The electron microscopy and spectroscopy tools confirmed the enzyme immobilization process on magnetic nanoparticles. The immobilized cellulase exhibited filter paper, carboxymethyl cellulase and cellobiase activities of 11.82, 21.36 and 10.81 IU, respectively. The free and immobilized cellulase exhibited identical pH optima (pH 5.0) while different temperature optima of 50 °C and 60 °C, respectively. The immobilized enzyme retained 56.87% of its maximal activity after 6 h of pre-incubation at 60 °C. Km (Michaelis constant) and Vmax (maximum velocity) of immobilized enzyme were 11.76 mM and 1.17 μmol min−1 ml−1, respectively. The immobilized cellulase hydrolysed pre-treated rice straw with saccharification efficiency of 52.67%. Further, it could be reutilized for up to four saccharification cycles with retention of 50.34% activity. Therefore, the improved properties of magnetic nanoparticle-immobilized cellulase and its reusability benefits offer a promising potential for industrial production of fermentable sugars and ethanol from rice straw.