Enzyme Recovery Research Articles

Engineering proteinaceous colloidosomes as enzyme carriers for efficient and recyclable Pickering interfacial biocatalysis.

Despite Pickering interfacial biocatalysis being a popular topic in biphasic biocatalysis, the development of water-in-oil (w/o) emulsion systems stabilized by single particles remains a challenge. For the first time, hydrophobized proteinaceous colloidosomes with magnetic-responsiveness are developed to function as both an enzyme carrier and emulsifier, achieving a breakthrough in protein-based w/o Pickering bioconversion. Enzyme-loaded protein colloidosomes are synthesized by a facile and mild method via emulsion templating. This system exhibits superior catalytic activity to other systems at the oil–water interface. Besides, feasible enzyme recovery and reusability ensure that this novel system can be employed as an efficient and eco-friendly recyclable platform.

Metal ions coordinated immobilization of phenylalanine dehydrogenase by GO‐PEI with high activity recovery and enhanced stability

AbstractBACKGROUNDDehydrogenases dependent on NAD(P)H are widely applied for biosynthesis of chiral chemicals. However, they are unstable and easy denaturated compared with hydrolases. A new strategy for dehydrogenases immobilization with the advantages of easy operation and high activity recovery should be developed. Biocompatible hybrid materials (GO‐PEI‐M) have been obtained by metal ions coordinated polyethylenimine (PEI) functionalized graphene oxide (GO).RESULTSThe immobilized enzyme recovery rates of GO‐PEI‐Mg (magnesium ion coordinated GO‐PEI) and GO‐PEI‐Mn (manganese ion coordinated GO‐PEI). In addition, the stability of GO‐PEI‐Mn‐PheDH (Phenylalanine Dehydrogenase) and GO‐PEI‐Mg‐PheDH was greatly improved, and it maintained 65% and 79% activity, respectively, after seven times reuse. In addition, the stability of GO‐PEI‐Mn‐PheDH (Phenylalanine Dehydrogenase) and GO‐PEI‐Mg‐PheDH was greatly improved. After seven times reuse, GO‐PEI‐Mn‐PheDH and GO‐PEI‐Mn‐PheDH and maintained 65% and 79% activity, respectively. An assembly mechanism was proposed from the perspectives of multi‐level interactions, including hydrogen bonding, electrostatic forces, and coordinated bonds.CONCLUSIONGO‐PEI‐Metal ion hybrid materials offer a promising carrier for enzyme immobilization with the advantages of biocompatibility and relatively strong interaction. © 2020 Society of Chemical Industry

The effects of two corn-expressed phytases on broiler growth performance and tibia mineralization

Summary This study examined the effects of 2 corn-expressed phytases, differing in enzyme concentration within the grain (3,000 or 9,677 Fytase unit [FTU]/g), on broiler performance and tibia mineralization when used as a component in pelleted feed. Dietary treatments included a positive control (PC), with industry-recommended levels of calcium and phosphorous, and a negative control (NC), lower in calcium and nonphytate phosphorous. Six diets containing corn-expressed phytase at high or low grain concentrations, formulated to 3,000, 6,000, or 9,000 FTU/kg, were created using the NC. Diets were conditioned at 70°C for 15 s and pelleted using a 4.76 × 38.1 mm pellet die. Descriptive enzyme recovery after pelleting (58–360%) and mixer CV based on enzyme activity (8–24%) demonstrated variability with the phytase assay and did not show trends across treatment main effects. Diets were fed to Hubbard × Ross 708 (n = 1,632) male broilers for 39 d. On day 21, five birds/pen were euthanized, and tibiae were excised to determine tibia mineralization. Treatments were arranged in a 2 (corn-phytase product) × 3 (phytase inclusion) factorial in a randomized complete block design. Broilers fed the NC had increased day 0–39 feed conversion ratio (FCR) compared with all other treatments (P

Diallyl Sulfide-Mediated Modulation of the Fatty Acid Synthase (FASN) Leads to Cancer Cell Death in BaP-Induced Lung Carcinogenesis in Swiss Mice.

PurposeDiallyl sulfide (DAS), one of the organo-sulfur secondary metabolites in garlic, has been shown to inhibit the proliferation of cancer cells. The present study aimed to evaluate the mechanism of DAS in the prevention of benzo[a]pyrene (BaP)-induced lung cancer in a murine model.Materials and MethodsThe mice were exposed to 50 mg/kg of BaP twice a week for 4 weeks in order to induce lung carcinoma. Pretreatment of mice with DAS (100 mg/kg) was started 2 weeks before BaP exposure and further continued for 21 weeks. The effect of DAS and BaP was evaluated by studying various parameters in the serum and tissues of the treated or untreated BaP-exposed mice.ResultsThe histopathological findings demonstrated that DAS prevented the progression of malignant lung cancer and metastasis in the liver. A significant drop was observed in BaP-induced tumor marker enzymes (ADA, AHH, γ-GT, LDH) in the serum of the mice treated with DAS. Moreover, DAS treatment resulted in the recovery of antioxidant enzymes, SOD and CAT, in BaP-exposed mice. The induction of apoptosis and the destruction of cellular ROS were detected in cancer cells from the mice pre-treated with DAS. The immunohistochemical analysis revealed the up-regulation of fatty acid synthase (FASN) in the lungs and liver tissues of BaP-exposed mice and the treatment with DAS inhibited FASN expression.ConclusionThe findings of the present study indicated that DAS-induced apoptosis is strongly associated with the downregulation of FASN in tumor tissues. To the best of our knowledge, this is the first study that describes the role of FASN in BaP-induced lung carcinogenesis.

Hydroxyapatite-CoFe2O4 Magnetic Nanoparticle Composites for Industrial Enzyme Immobilization, Use, and Recovery

Hydroxyapatite (HA) is an inorganic material with high ability to interact with proteins and has been recently explored as a support for enzyme immobilization. However, there are still some drawbacks concerning the recovery of these biocatalysts, which could be overcome using magnetic supports. Cobalt ferrite (CoFe2O4) is a purely magnetic material, which offers excellent chemical stability, ease of synthesis, and mechanical hardness, being a promising candidate to form composites with HA and produce magnetic HA nanoparticles. Therefore, investigation of synthesis procedures and applications of HA/CoFe2O4 composites to facilitate enzyme recovery by means of a magnetic field could find interest in a broad spectrum of biotechnological processes. Here, the co-precipitation method was used to synthesize HA/CoFe2O4 composites with different HA/CoFe2O4 mass ratios, for application as supports for enzyme immobilization. The enzymes β-glucosidase, phytase, and xylanase were selected for proof of concept due to their wide range of industrial applications. The composite with the highest cobalt ferrite content (2:1 mass ratio) was highly effective for immobilization of the three different enzymes, with immobilization yields (IYs) of 70–100% and recovered activities of 78–100%. The biocatalysts could be easily recovered from the reaction media by both centrifugation and application of an external magnetic field, demonstrating their potential for use in industrial processes. The materials exhibited good reusability, especially in the case of the β-glucosidase biocatalyst, which could be reused 10 times, maintaining around 70% of its initial activity.

Momordica charantia Extract Protects against Diabetes-Related Spermatogenic Dysfunction in Male Rats: Molecular and Biochemical Study

More than 90% of diabetic patients suffer from sexual dysfunction, including diminished sperm count, sperm motility, and sperm viability, and low testosterone levels. The effects of Momordica charantia (MC) were studied by estimating the blood levels of insulin, glucose, glycosylated hemoglobin (HbA1c), testosterone (TST), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) in diabetic rats treated with 250 and 500 mg/kg b.w. of the total extract. Testicular antioxidants, epididymal sperm characteristics, testicular histopathology, and lesion scoring were also investigated. Testicular mRNA expression of apoptosis-related markers such as antiapoptotic B-cell lymphoma-2 (Bcl-2) and proapoptotic Bcl-2-associated X protein (Bax) were evaluated by real-time PCR. Furthermore, caspase-3 protein expression was evaluated by immunohistochemistry. MC administration resulted in a significant reduction in blood glucose and HbA1c and marked elevation of serum levels of insulin, TST, and gonadotropins in diabetic rats. It induced a significant recovery of testicular antioxidant enzymes, improved histopathological changes of the testes, and decreased spermatogenic and Sertoli cell apoptosis. MC effectively inhibited testicular apoptosis, as evidenced by upregulation of Bcl-2 and downregulation of Bax and caspase-3. Moreover, reduction in apoptotic potential in MC-treated groups was confirmed by reduction in the Bax/Bcl-2 mRNA expression ratio.

Recycling of Chrysoporthe cubensis enzymes to overcome the adsorption in lignin and to improve sugarcane bagasse saccharification

Biomass enzymatic hydrolysis efficiency shows some limitations, such as the non-specific adsorption of enzymes in lignin. We evaluated the adsorption of proteins and enzymes from Chrysoporthe cubensis into alkaline and acid pretreated sugarcane bagasse, containing 8.1 and 33.8% of lignin, respectively. The data from Langmuir isotherm showed that at 4 °C, proteins were more adsorbed by acid-treated bagasse, 10.5 mg g−1, while higher adsorption affinity, 2.0 mL mg−1, was determined from alkali-treated bagasse. Similarly, under hydrolytic conditions (50 °C), higher amounts of proteins were adsorbed into acid-treated bagasse, confirming the predominant effect of lignin on protein adsorption. Enzymatic recycling rounds during alkali-treated bagasse saccharification showed that the addition of fresh substrate and enzymes promoted the higher release of glucose and xylose, 40.3 and 38.3 g L−1, respectively. However, the addition of only fresh substrates showed to be efficient for C. cubensis enzymes recovery and substrate hydrolysis, which reduces enzyme consumption and costs.

Aqueous Two-Phase Systems for Cleanup and Recovery of Enzymes from Plants and Plant-Derived Extracts.

The increasing interest of the biopharmaceutical industry to exploit plants as a commercially viable production system is demanding the development of new strategies to maximize product recovery. Aqueous two-phase systems (ATPSs) are a primary recovery technique that has shown great potential for the efficient extraction and purification of biological products, from organelles to proteins and low-molecular-weight compounds. The evaluation of different system parameters upon the partitioning behavior can provide the conditions that favor the concentration of contaminants and the desired target protein in opposite phases. The protocols described here provide the basic strategy to explore the use of ATPSs for the isolation and partial purification of native and recombinant proteins from plants and plant-derived extracts.

Zbasic: A Purification Tag for Selective Ion-Exchange Recovery.

A positively charged protein domain, denoted Zbasic, can be used as a general purification tag for purification of recombinantly produced target proteins by cation-exchange chromatography. The Zbasic domain is constructed from the Protein A-derived Z-domain, and engineered to be highly charged, which allows selective capture on a cation exchanger at physiological pH values. Moreover, Zbasic is selective also under denaturing conditions and can be used for purification of proteins solubilized from inclusion bodies. Zbasic can then be used as a flexible linker to the cation-exchanger resin, and thereby allows solid-phase refolding of the target protein.Herein, protocols for purification of soluble Zbasic-tagged fusion proteins , as well as for integrated purification and solid-phase refolding of insoluble fusion proteins , are described. In addition, a procedure for enzymatic tag removal and recovery of native target protein is outlined.

Tubercular splenic abscess and recurrent acute on chronic tubercular pancreatitis: a rare coincidence in an immune competent child

Tubercular splenic abscess in association with chronic tubercular pancreatitis is scarcely reported in an immune competent child. 12 years old boy had presented with recurrent attacks of upper abdominal pain radiating towards left shoulder. He had multiple hospitalizations with similar complaints. The child had associated anorexia and significant weight loss during the period of his illness. Clinical examination revealed tenderness and muscle guard over the epigastric region. The pancreatic enzymes were markedly raised. Abdominal CT scan had suggested chronic pancreatitis with multiple splenic abscesses. The cartridge- based nucleic acid amplification test (CBNAAT) of the pus aspirated from the abscess confirmed the presence of pan sensitive mycobacterium tuberculosis. Image guided fine needle aspiration biopsy revealed the presence of caseating granulomatous lesion. Treatment with anti-tubercular drugs led to complete clinical recovery and normalization of pancreatic enzymes at follow up. An immunocompetent child had chronic tubercular pancreatitis associated with tubercular splenic abscess.

Current Developments in Lignocellulosic Biomass Conversion into Biofuels Using Nanobiotechology Approach

The conversion of lignocellulosic biomass (LB) to sugar is an intricate process which is the costliest part of the biomass conversion process. Even though acid/enzyme catalysts are usually being used for LB hydrolysis, enzyme immobilization has been recognized as a potential strategy nowadays. The use of nanobiocatalysts increases hydrolytic efficiency and enzyme stability. Furthermore, biocatalyst/enzyme immobilization on magnetic nanoparticles enables easy recovery and reuse of enzymes. Hence, the exploitation of nanobiocatalysts for LB to biofuel conversion will aid in developing a lucrative and sustainable approach. With this perspective, the effects of nanobiocatalysts on LB to biofuel production were reviewed here. Several traits, such as switching the chemical processes using nanomaterials, enzyme immobilization on nanoparticles for higher reaction rates, recycling ability and toxicity effects on microbial cells, were highlighted in this review. Current developments and viability of nanobiocatalysts as a promising option for enhanced LB conversion into the biofuel process were also emphasized. Mostly, this would help in emerging eco-friendly, proficient, and cost-effective biofuel technology.

The effect of the level of mixer-added water and mash conditioning temperature on parameters monitored during pelleting and phytase and xylanase thermostability

Exogenous enzymes have been used in broiler and swine rations to improve nutrient utilization and reduce feed costs, both of which improved the efficiencies of live production. Feed manufacturers have also utilized practices, such as warm mash conditioning temperatures (CT) and the addition of moisture at the mixer or mash conditioning chamber, to maintain adequate physical feed quality. While the impact of warm conditioning practices on enzyme thermostability had been well described, the impact of mixer-added water (MAW) on enzyme thermostability was scarcely investigated in previous literature. The present experiment intended to investigate the interaction of three mash CT and three levels of MAW on the thermostability of a phytase and xylanase mixer-added enzyme within a swine gestation diet in a 3 × 3 factorial randomized complete block design. The mash CT included 80, 86, and 92 °C and the levels of MAW included 0, 10, and 20 g/kg. Parameters monitored during the pelleting process included pellet durability index (PDI), pellet mill energy consumption (PMEC), and the change in temperature between hot pellets and conditioned mash (Δ T). Conditioning mash at 92 °C resulted in a 22.5 percentage-point reduction in phytase recovery in conditioned mash and a 70.1 percentage-point reduction in phytase recovery in pellets when compared to diets pelleted at 80 °C (P < 0.01). The xylanase, however, was not affected by mash CT. Phytase and xylanase recovery in pellets was not affected by the level of MAW. As PDI and PMEC increased, phytase recovery in pellets relative to unconditioned mash decreased (P < 0.01). As Δ T increased, phytase recovery in pellets relative to unconditioned mash increased (P < 0.01). Xylanase denaturation was poorly described by the parameters monitored during pelleting, likely because it was minimally denatured during pelleting. A robust multiple regression model was generated to predict phytase activity in pellets relative to unconditioned mash (R2 = 0.95; P < 0.01) when the controlled factors and monitored parameters were combined in a predictive model. In conclusion, the primary site of enzyme denaturation in a pilot scale pellet mill was the pellet mill die and MAW did not affect enzyme recovery.

Determination of acute median lethal concentration and sublethal effects on AChE activity of Gymnotus carapo (Teleostei: Gymnotidae) exposed to trichlorfon

Trichlorfon (TRF) is a pesticide widely used in aquaculture to control fish ectoparasites. This pesticide is an inhibitor of acetylcholinesterase, an essential enzyme for termination of nerve impulses. High rates of TRF use generate risks to the environment and human health. In the environment, pesticides can affect the local fauna and generate an ecological breakdown. There are several studies performed with fish production; however, gaps are created for native fish with other commercial values. The tuvira (Gymnotus carapo) is a fish native to Brazilian fauna and has great commercial importance in sport fishing. The present study aimed to determine the lethal concentration of trichlorfon (Masoten) in Gymnotus carapo and its sublethal effects on the enzyme AChE. In this study, the acute toxicity (the concentrations to kill 50% of the fish LC50) of TRF in tuviras (Gymnotus carapo) and acetylcholinesterase inhibition in liver and muscle tissue of tuviras submitted to sublethal concentrations were evaluated. For the acute assay, concentrations of 0.0, 5.0, 7.5, 15, 22.5, 30, 37.5 and 45 mg L-1 were used for a period of 96 h. After the acute exposure period, a LC50 of 6.38 mg L-1 was determined. In the sublethal assay, concentrations of 0.0, 0.238, 0.438 and 0.638 mg L-1 were used, based on 10% of the LC50, over a period of 14 days. Two collections were performed: one at seven days and the other at the end (day 14). Inhibition of acetylcholinesterase in the liver was only shown (p &lt; 0.05) for the treatment with 0.638 mg L-1 after 14 days of exposure. At seven days, muscle activity showed a significant difference only for the treatments 0.438 and 0.638 mg L-1, compared with the treatment 0.238 mg L-1 and control. At 14 days of exposure, only the treatment 0.638 mg L-1 showed significant differences in relation to the other groups, thus showing that enzyme recovery had occurred. The value found in the acute test allowed the conclusion that TRF presents moderately toxic characteristics to Gymnotus carapo. The toxicity parameter values calculated in the present study assisted in estimation of maximum allowable limits in bodies of water when combined with test data from other non-target organisms.

Purification and thrombolytic effects in vivo of recombination Nattokinase on carrageenan-induced tail thrombosis in a rat model

Nattokinase is a serine protease with fibrinolytic activity and has been proven clinical efficacy and safety for human use by the oral route. In this study, conducted purified recombinant nattokinase from Bacillus subtillis&#x0D; DB104 strain and thrombolytic effects of nattokinase have been observed in vivo. The result of the enzyme purification by membrane filtration method for enzyme recovery efficiency is 82.45%. Then the enzyme was purified to Q-Sepharose ion exchange chromatography with a purification factor of 1.82 and at a yield of 66.91%. Enzyme activity was 5,457 FU/mg protein. With the carrageenan-induced tail thrombosis in a rat model, results were significantly reduced by 67.3 to 83.6% at oral doses and injected nattokinase compared with the control group at 48 h. At 72 hours the rate increased to 82-89% in the oral dose and up to 91% in the injected dose nattokinase and comparable to 90% for heparin-positive hepatitis.

Magnetically recyclable catalytic nanoparticles grafted with Bacillus subtilis β-glucosidase for efficient cellobiose hydrolysis

This study reports covalent immobilization of β-glucosidase (BGL) from Bacillus subtilis PS on magnetically recyclable iron nanoparticles for enhancing robustness, facile recovery and reuse of enzyme. Immobilized BGL iron nanoparticles (BGL-INPs) were characterized by various biophysical techniques viz. TEM, DLS, FTIR and CD spectroscopy. The efficiency and yield of immobilization were 89.78 and 84.80%, respectively. After immobilization, optimum pH remained 6.0 whereas optimum temperature upraised to 70 °C whereas apparent Km and Vmax shifted from 0.819 mM to 0.941 mM and 54.46 to 57.67 μmole/min/mg, respectively. Immobilization conferred lower activation energy and improved pH and thermal stabilities. The BGL-INPs retained 85% activity up to 10th cycle of reuse and hydrolyzed more than 90% of cellobiose to glucose within 30 min. Conclusively, improved pH, thermal stability and excellent reusability over free enzyme make BGL-INPs a promising candidate for sustainable bioethanol production and other industrial applications.

Efficient production of fructo-oligosaccharides from sucrose and molasses by a novel Aureobasidium pullulan strain

The use of whole-cell biocatalyst for production of fructo-oligosaccharide (FOS) eliminates the need for costly enzyme recovery and purification. In this study, a novel Aureobasidium pullulans strain (FRR 5284) was identified from seven A. pulllulans strains as an efficient whole-cell biocatalyst for FOS production. The strain had a specific intracellular transfructosylating activity of 4.44 ± 0.18 U/mg dry cells, one of the highest transfructosylating activities thus far reported for whole-cell biocatalyst. Under optimal conditions (pH 5.5 and 55 °C), only 5 g/L of dry cells and 3 h reaction time were required to achieve a FOS yield of 61 % from 50 % (w/v) sucrose. Incubation of sugarcane molasses with an invertase-free Saccharomyces cerevisiae prior to addition of A. pullulans whole-cell biocatalyst eliminated glucose inhibition and increased FOS yield from 44 % to 56 % in 1 h. This study has demonstrated that the novel A. pullulans FRR 5284 was an efficient source of whole-cell biocatalyst for FOS production and a promising strategy for transformation of sucrose and sugarcane molasses into a higher-value prebiotic.

Study of Microbial Transglutaminase Partitioning in Thermo-pH-Responsive Aqueous Two-Phase Systems.

The recyclable aqueous two-phase systems (ATPS) responding to environmental stimuli have been widely studied in the purification of biologics. In this study, a thermo-responsive polymer PNE was copolymerized after optimization of monomer ratio. In addition, its lower critical solution temperature (LCST, 31°C) and first recovery (99.43%) were determined. Then, PNE was used to form two recyclable ATPS with another thermo-responsive polymer PNDBN and a pH-responsive polymer PADB4.91, which were polymers already prepared in the lab. Meanwhile, the partition behavior of microbial transglutaminase (MTG) was explored using these two ATPS. The result showed that the PNE/PADB4.91 ATPS was superior to PNDBN/PNE ATPS owing to its faster phase formation and better partition performance. In order to optimize the partition behavior, several parameters were investigated based on PNE/PADB4.91 ATPS. In the ATPS constructed with 2.5% (w/v) PNE and 3.5% (w/v) PADB4.91, the maximal partition coefficient (1/KE) and enzymatic recovery (ERB) of MTG were 12.9 and 95.21% in the presence of 10mM KCl when the temperature, pH, and the addition amount of MTG were 25°C, 7.0, and 2mg/mL, respectively.

Utilization of biosolids for glucose oxidase production: A potential bio-fenton reagent for advanced oxidation process for removal of pharmaceutically active compounds

The current work focuses on the production of glucose oxidase (GOD) in sterilized biosolid (BS) slurries containing BS and municipal wastewater effluent. Various parameters were optimized for maximizing the GOD production and the effects of biostimulation on GOD production was investigated by adding synthetic media components. The studies on inoculum characteristics at an inoculum age of 72 h and inoculum size of 20% (w/v) produced high GOD activities of around 6012 U/L in 25% (dw/v) BS media. Further, the effect of ultrasonication time was determined to release BS-bound GOD in order to maximize enzymes recovery. Using 1000 U/L of the BS-based GOD for 0.55 M glucose oxidation produced the maximum H2O2 concentration of 216 ppm. The produced H2O2 was utilized for bio-Fenton based advanced oxidation process for the partial removal of 15 pharmaceutically active compounds.

Analysis of clinical characteristics of tigecycline-induced acute pancreatitis.

The purpose of this study is to explore the clinical characteristics of tigecycline-induced acute pancreatitis. We searched the PubMed/Medline, Web of Knowledge, OVID, Elsevier, Springer Link, Embase, Cochrane Library, China National Knowledge Infrastructure (CNKI), Wanfang Data, and Chinese VIP databases from 2005 and identified 19 studies of tigecycline-induced acute pancreatitis involving a total of 22 patients for inclusion in a retrospective analysis. The median (range)age of 22 patients with acute pancreatitis was 58years (range 9-83). Overall, the median (range) time of symptom onset was 6.5days (range 2-28), or 6days (range 2-14) and 6days (range 3-28) in patients with or without a loading dose of tigecycline, respectively. Symptoms included nausea, vomiting and abdominal distension (73%) and abdominal pain (73%); 90% (18/20) of patients developed mild acute pancreatitis (MAP), and 10% (2/20) developed severe acute pancreatitis (SAP). Computed tomography (CT) scans showed oedematous infiltrate in 56% (10/18) of cases and acute pancreatitis in 28% (5/18) of cases. The median (range) level of lipase and amylase was 936U/L (range 382-4089) and 588U/L (range 312-1166), respectively. The median (range) time to recovery of symptoms was 4days (range 1-10), and the time for recovery of pancreatic enzymes to the normal range was 5days (range 1-30) after the withdrawal of tigecycline in all patients. Clinicians should be particularly mindful of clinical signs and symptoms, the level of serum pancreatic enzymes and abdominal CT images in order to monitor the development of pancreatitis when using tigecycline.

Maximizing the direct recovery and stabilization of cellulolytic enzymes from Trichoderma harzanium BPGF1 fermented broth using carboxymethyl inulin nanoparticles

The carboxymethylated inulin (CMI) nanoparticles prepared by the salt out method was demonstrated to harvest cellulolytic enzymes (Ez) directly from the clarified fermented broth of Trichoderma harzanium BPGF1. The formation of CMI nanoparticles and entrapment of Ez in CMI was confirmed by scanning electron microscopy and Fourier transform infrared spectroscopy, respectively. A factorial design was developed to maximize enzymes recovery directly from the fermented broth. A maximum of 71.68 ± 8.61% cellulolytic enzymes was recovered using 20 mg/L inulin, 2 M sodium chloroacetate at 80 °C for 2 h. The resultant CMIEz nanohybrid displayed excellent activity in broad pH and temperature. Moreover, CMIEz was reusable for >30 cycles without losing efficiency. The real-time application of CMIEz was demonstrated by hydrolyzing acid pretreated corncob. High-pressure liquid chromatography revealed that the hydrolyzed corncob contained cellobiose, glucose, galactose, xylose, mannose, and arabinose. The results highlight that carbohydrate nanoparticles was useful in engulfing enzymes directly from the fermentation broth.