The Experts below are selected from a list of 116310 Experts worldwide ranked by ideXlab platform
Shah Ali Ul Qader - One of the best experts on this subject based on the ideXlab platform.
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maltase entrapment approach as an efficient alternative to increase the stability and recycling efficiency of Free Enzyme within agarose matrix
Journal of The Taiwan Institute of Chemical Engineers, 2016Co-Authors: Afsheen Aman, Haneef Ur Rehman, Dilshad Hussain, Midrar Ullah, Asad Karim, Zainab Bibi, Muhammad Nawaz, Shah Ali Ul QaderAbstract:Abstract Maltase catalyses the hydrolysis of maltose and is widely used in the synthesis of various food and pharmaceutical products. In the current study, matrix entrapment technique was applied to immobilize maltase within agarose beads. Maximum immobilization yield (76.11%) was achieved at 3% agarose concentration and 5.0 mm beads size was found to be optimum combination for maximum catalytic activity of entrapped maltase. It was noticed that entrapment increased the optimum reaction temperature and pH of maltase from 45°C to 60°C and 6.5 to 7.0, respectively with reference to its Free Enzyme whereas no effect was observed on reaction time. Entrapped maltase showed increase in K m value from 1.717 to 1.912 mM ml −1 and decrease in V max value from 8411.0 to 6214.0 U ml −1 min −1 as compared to Free Enzyme. Entrapped maltase displayed broad thermal stability up to 80°C whereas; Free Enzyme completely lost it activity when temperature reached up to 60°C. Scanning electron microscopy of agarose beads before and after maltase entrapment revealed significant morphological change on the matrix surface. Considering the economic feasibility, the entrapped maltase indicated imperative recycling efficiency up to ten reaction cycles.
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continuous degradation of maltose improvement in stability and catalytic properties of maltase α glucosidase through immobilization using agar agar gel as a support
Bioprocess and Biosystems Engineering, 2015Co-Authors: Muhammad Nawaz, Afsheen Aman, Shah Ali Ul Qader, Asad Karim, Roberta Marchetti, Antonio MolinaroAbstract:Maltose degrading Enzyme was immobilized within agar-agar support via entrapment method due to its industrial utilization. The maximum immobilization efficiency (82.77 %) was achieved using 4.0 % agar-agar keeping the diameter of bead up to 3.0 mm. The matrix entrapment showed maximum catalytic activity at pH 7.0 and temperature 65 °C. Substrate saturation kinetics showed that the K m of immobilized Enzyme increased from 1.717 to 2.117 mM ml−1 where as Vmax decreased from 8,411 to 7,450 U ml−1 min−1 as compared to Free Enzyme. The immobilization significantly increased the stability of maltase against various temperatures and immobilized maltase retain 100 % of its original activity after 2 h at 50 °C, whereas the Free maltase only showed 60 % residual activity under same condition. The reusability of entrapped maltase showed activity up to 12 cycles and retained 50 % of activity even after 5th cycle. Storage stability of agar entrapped maltase retain 73 % of its initial activity even after 2 months when stored at 30 °C while Free Enzyme showed only 37 % activity at same storage conditions. .
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agar agar entrapment increases the stability of endo β 1 4 xylanase for repeated biodegradation of xylan
International Journal of Biological Macromolecules, 2015Co-Authors: Zainab Bibi, Shah Ali Ul Qader, Faiza Shahid, Afsheen AmanAbstract:Abstract Microbial xylanases, specially endo-β-1,4-xylanase catalyzes the hydrolysis of xylan, is considered one of the most significant hydrolases. It has numerous applications but most extensively is utilized in paper and pulp industry as a bio-bleaching agent. Immobilization technique is comprehensively studied with the expectation of modifying and improving Enzyme stability and characteristics for commercial purposes. Currently, matrix entrapment technique is applied to immobilize endo-β-1,4-xylanase within agar–agar gel beads produced by Geobacillus stearothermophilus KIBGE-IB29. Maximal Enzyme immobilization yield was achieved at 2.5% of agar–agar concentration. Optimized conditions demonstrated an increase in the optimal reaction time from 05 min to 30 min and incubation temperature from 50 °C to 60 °C with reference to Free Enzyme whereas; no effect was observed for optimum pH. Entrapment technique uniquely changed the kinetic parameters of immobilized endo-β-1,4-xylanase ( K m : 0.5074 mg min −1 to 0.5230 mg min −1 and V max : 4773 U min −1 to 968 U min −1 ) as compared to Free Enzyme. However, immobilized Enzyme displayed broad thermal stability and retained 79.0% of its initial activity at 80 °C up to 30 min whereas; Free Enzyme completely lost its activity at this temperature. With respect to economic feasibility, the immobilized Enzyme showed impressive recycling efficiency up to six reaction cycles.
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immobilization of pectin degrading Enzyme from bacillus licheniformis kibge ib 21 using agar agar as a support
Carbohydrate Polymers, 2014Co-Authors: Haneef Ur Rehman, Afsheen Aman, Raheela Rahmat Zohra, Shah Ali Ul QaderAbstract:Abstract Pectinase from Bacillus licheniformis KIBGE IB-21 was immobilized in agar-agar matrix using entrapment technique. Effect of different concentrations of agar-agar on pectinase immobilization was investigated and it was found that maximum immobilization was achieved at 3.0% agar-agar with 80% Enzyme activity. After immobilization, the optimum temperature of Enzyme increased from 45 to 50 °C and reaction time from 5 to 10 minutes as compared to Free Enzyme. Due to the limited diffusion of high molecular weight substrate, K m of immobilized Enzyme slightly increased from 1.017 to 1.055 mg ml −1 , while V max decreased from 23,800 to 19,392 μM min −1 as compared to Free Enzyme. After 120 h entrapped pectinase retained their activity up to 82% and 71% at 30 °C and 40 °C, respectively. The entrapped pectinase showed activity until 10th cycle and maintain 69.21% activity even after third cycle.
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degradation of complex carbohydrate immobilization of pectinase from bacillus licheniformis kibge ib21 using calcium alginate as a support
Food Chemistry, 2013Co-Authors: Haneef Ur Rehman, Afsheen Aman, Shah Ali Ul Qader, Antonio Molinaro, Alba Silipo, Asma AnsariAbstract:Abstract Pectinases are heterogeneous group of Enzymes that catalyse the hydrolysis of pectin substances which is responsible for the turbidity and undesirable cloudiness in fruits juices. In current study, partially purified pectinase from Bacillus licheniformis KIBGE-IB21 was immobilized in calcium alginate beads. The effect of sodium alginate and calcium chloride concentration on immobilization was studied and it was found that the optimal sodium alginate and calcium chloride concentration was 3.0% and 0.2 M, respectively. It was found that immobilization increases the optimal reaction time for pectin degradation from 5 to 10 min and temperature from 45 to 55 °C, whereas, the optimal pH remained same with reference to Free Enzyme. Thermal stability of Enzyme increased after immobilization and immobilized pectinase retained more than 80% of its initial activity after 5 days at 30 °C as compared with Free Enzyme which showed only 30% of residual activity. The immobilized Enzyme also exhibited good operational stability and 65% of its initial activity was observed during third cycle. In term of pectinase immobilization efficiency and stability, this calcium alginate beads approach seemed to permit good results and can be used to make a bioreactor for various applications in food industries.
Afsheen Aman - One of the best experts on this subject based on the ideXlab platform.
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maltase entrapment approach as an efficient alternative to increase the stability and recycling efficiency of Free Enzyme within agarose matrix
Journal of The Taiwan Institute of Chemical Engineers, 2016Co-Authors: Afsheen Aman, Haneef Ur Rehman, Dilshad Hussain, Midrar Ullah, Asad Karim, Zainab Bibi, Muhammad Nawaz, Shah Ali Ul QaderAbstract:Abstract Maltase catalyses the hydrolysis of maltose and is widely used in the synthesis of various food and pharmaceutical products. In the current study, matrix entrapment technique was applied to immobilize maltase within agarose beads. Maximum immobilization yield (76.11%) was achieved at 3% agarose concentration and 5.0 mm beads size was found to be optimum combination for maximum catalytic activity of entrapped maltase. It was noticed that entrapment increased the optimum reaction temperature and pH of maltase from 45°C to 60°C and 6.5 to 7.0, respectively with reference to its Free Enzyme whereas no effect was observed on reaction time. Entrapped maltase showed increase in K m value from 1.717 to 1.912 mM ml −1 and decrease in V max value from 8411.0 to 6214.0 U ml −1 min −1 as compared to Free Enzyme. Entrapped maltase displayed broad thermal stability up to 80°C whereas; Free Enzyme completely lost it activity when temperature reached up to 60°C. Scanning electron microscopy of agarose beads before and after maltase entrapment revealed significant morphological change on the matrix surface. Considering the economic feasibility, the entrapped maltase indicated imperative recycling efficiency up to ten reaction cycles.
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continuous degradation of maltose improvement in stability and catalytic properties of maltase α glucosidase through immobilization using agar agar gel as a support
Bioprocess and Biosystems Engineering, 2015Co-Authors: Muhammad Nawaz, Afsheen Aman, Shah Ali Ul Qader, Asad Karim, Roberta Marchetti, Antonio MolinaroAbstract:Maltose degrading Enzyme was immobilized within agar-agar support via entrapment method due to its industrial utilization. The maximum immobilization efficiency (82.77 %) was achieved using 4.0 % agar-agar keeping the diameter of bead up to 3.0 mm. The matrix entrapment showed maximum catalytic activity at pH 7.0 and temperature 65 °C. Substrate saturation kinetics showed that the K m of immobilized Enzyme increased from 1.717 to 2.117 mM ml−1 where as Vmax decreased from 8,411 to 7,450 U ml−1 min−1 as compared to Free Enzyme. The immobilization significantly increased the stability of maltase against various temperatures and immobilized maltase retain 100 % of its original activity after 2 h at 50 °C, whereas the Free maltase only showed 60 % residual activity under same condition. The reusability of entrapped maltase showed activity up to 12 cycles and retained 50 % of activity even after 5th cycle. Storage stability of agar entrapped maltase retain 73 % of its initial activity even after 2 months when stored at 30 °C while Free Enzyme showed only 37 % activity at same storage conditions. .
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agar agar entrapment increases the stability of endo β 1 4 xylanase for repeated biodegradation of xylan
International Journal of Biological Macromolecules, 2015Co-Authors: Zainab Bibi, Shah Ali Ul Qader, Faiza Shahid, Afsheen AmanAbstract:Abstract Microbial xylanases, specially endo-β-1,4-xylanase catalyzes the hydrolysis of xylan, is considered one of the most significant hydrolases. It has numerous applications but most extensively is utilized in paper and pulp industry as a bio-bleaching agent. Immobilization technique is comprehensively studied with the expectation of modifying and improving Enzyme stability and characteristics for commercial purposes. Currently, matrix entrapment technique is applied to immobilize endo-β-1,4-xylanase within agar–agar gel beads produced by Geobacillus stearothermophilus KIBGE-IB29. Maximal Enzyme immobilization yield was achieved at 2.5% of agar–agar concentration. Optimized conditions demonstrated an increase in the optimal reaction time from 05 min to 30 min and incubation temperature from 50 °C to 60 °C with reference to Free Enzyme whereas; no effect was observed for optimum pH. Entrapment technique uniquely changed the kinetic parameters of immobilized endo-β-1,4-xylanase ( K m : 0.5074 mg min −1 to 0.5230 mg min −1 and V max : 4773 U min −1 to 968 U min −1 ) as compared to Free Enzyme. However, immobilized Enzyme displayed broad thermal stability and retained 79.0% of its initial activity at 80 °C up to 30 min whereas; Free Enzyme completely lost its activity at this temperature. With respect to economic feasibility, the immobilized Enzyme showed impressive recycling efficiency up to six reaction cycles.
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immobilization of pectin degrading Enzyme from bacillus licheniformis kibge ib 21 using agar agar as a support
Carbohydrate Polymers, 2014Co-Authors: Haneef Ur Rehman, Afsheen Aman, Raheela Rahmat Zohra, Shah Ali Ul QaderAbstract:Abstract Pectinase from Bacillus licheniformis KIBGE IB-21 was immobilized in agar-agar matrix using entrapment technique. Effect of different concentrations of agar-agar on pectinase immobilization was investigated and it was found that maximum immobilization was achieved at 3.0% agar-agar with 80% Enzyme activity. After immobilization, the optimum temperature of Enzyme increased from 45 to 50 °C and reaction time from 5 to 10 minutes as compared to Free Enzyme. Due to the limited diffusion of high molecular weight substrate, K m of immobilized Enzyme slightly increased from 1.017 to 1.055 mg ml −1 , while V max decreased from 23,800 to 19,392 μM min −1 as compared to Free Enzyme. After 120 h entrapped pectinase retained their activity up to 82% and 71% at 30 °C and 40 °C, respectively. The entrapped pectinase showed activity until 10th cycle and maintain 69.21% activity even after third cycle.
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degradation of complex carbohydrate immobilization of pectinase from bacillus licheniformis kibge ib21 using calcium alginate as a support
Food Chemistry, 2013Co-Authors: Haneef Ur Rehman, Afsheen Aman, Shah Ali Ul Qader, Antonio Molinaro, Alba Silipo, Asma AnsariAbstract:Abstract Pectinases are heterogeneous group of Enzymes that catalyse the hydrolysis of pectin substances which is responsible for the turbidity and undesirable cloudiness in fruits juices. In current study, partially purified pectinase from Bacillus licheniformis KIBGE-IB21 was immobilized in calcium alginate beads. The effect of sodium alginate and calcium chloride concentration on immobilization was studied and it was found that the optimal sodium alginate and calcium chloride concentration was 3.0% and 0.2 M, respectively. It was found that immobilization increases the optimal reaction time for pectin degradation from 5 to 10 min and temperature from 45 to 55 °C, whereas, the optimal pH remained same with reference to Free Enzyme. Thermal stability of Enzyme increased after immobilization and immobilized pectinase retained more than 80% of its initial activity after 5 days at 30 °C as compared with Free Enzyme which showed only 30% of residual activity. The immobilized Enzyme also exhibited good operational stability and 65% of its initial activity was observed during third cycle. In term of pectinase immobilization efficiency and stability, this calcium alginate beads approach seemed to permit good results and can be used to make a bioreactor for various applications in food industries.
Guifang Jia - One of the best experts on this subject based on the ideXlab platform.
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antibody Free Enzyme assisted chemical approach for detection of n6 methyladenosine
Nature Chemical Biology, 2020Co-Authors: Ye Wang, Shunqing Dong, Guifang Jia, Yu XiaoAbstract:The inert chemical property of RNA modification N6-methyladenosine (m6A) makes it very challenging to detect. Most m6A sequencing methods rely on m6A-antibody immunoprecipitation and cannot distinguish m6A and N6,2′-O-dimethyladenosine modification at the cap +1 position (cap m6Am). Although the two antibody-Free methods (m6A-REF-seq/MAZTER-seq and DART-seq) have been developed recently, they are dependent on m6A sequence or cellular transfection. Here, we present an antibody-Free, FTO-assisted chemical labeling method termed m6A-SEAL for specific m6A detection. We applied m6A-SEAL to profile m6A landscapes in humans and plants, which displayed the known m6A distribution features in transcriptome. By doing a comparison with all available m6A sequencing methods and specific m6A sites validation by SELECT, we demonstrated that m6A-SEAL has good sensitivity, specificity and reliability for transcriptome-wide detection of m6A. Given its tagging ability and FTO’s oxidation property, m6A-SEAL enables many applications such as enrichment, imaging and sequencing to drive future functional studies of m6A and other modifications. An antibody-independent m6A profiling method called m6A-SEAL was developed via turning m6A into stable dm6A after treatment by FTO and DTT. This method exhibits better reliability in detection of transcriptome-wide m6A sites in humans and plants.
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antibody Free Enzyme assisted chemical labeling for detection of transcriptome wide n6 methyladenosine
bioRxiv, 2019Co-Authors: Ye Wang, Shunqing Dong, Qiong Yu, Yu Xiao, Guifang JiaAbstract:Abstract The inert chemical property of RNA modification N6-methyladenosine (m6A) makes it very challenging to detect, and all of the transcriptome-wide m6A detection methods rely on m6A-antibody immunoprecipitation. However, their results are dependent on the quality and specificity of antibodies. Although the endoribonuclease-based single-base m6A sequencing is antibody-Free, it maps only 16~25% sites. Here, we present an antibody-Free, FTO-assisted chemical labeling method termed m6A-SEAL for m6A detection. We applied m6A-SEAL to profile m6A landscapes in human and plant, which had good overlaps with antibody-based results and displayed the known m6A distribution features in transcriptome. Comparison with all available m6A sequencing methods and specific m6A sites validation by SELECT, we demonstrated that m6A-SEAL has good sensitivity, specificity, and reliability for transcriptome-wide detection of m6A. Given its tagging ability and FTO’s oxidation property, m6A-SEAL enables many applications like enrichment, imaging, and sequencing techniques to drive future functional studies of m6A and other modifications.
Haneef Ur Rehman - One of the best experts on this subject based on the ideXlab platform.
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maltase entrapment approach as an efficient alternative to increase the stability and recycling efficiency of Free Enzyme within agarose matrix
Journal of The Taiwan Institute of Chemical Engineers, 2016Co-Authors: Afsheen Aman, Haneef Ur Rehman, Dilshad Hussain, Midrar Ullah, Asad Karim, Zainab Bibi, Muhammad Nawaz, Shah Ali Ul QaderAbstract:Abstract Maltase catalyses the hydrolysis of maltose and is widely used in the synthesis of various food and pharmaceutical products. In the current study, matrix entrapment technique was applied to immobilize maltase within agarose beads. Maximum immobilization yield (76.11%) was achieved at 3% agarose concentration and 5.0 mm beads size was found to be optimum combination for maximum catalytic activity of entrapped maltase. It was noticed that entrapment increased the optimum reaction temperature and pH of maltase from 45°C to 60°C and 6.5 to 7.0, respectively with reference to its Free Enzyme whereas no effect was observed on reaction time. Entrapped maltase showed increase in K m value from 1.717 to 1.912 mM ml −1 and decrease in V max value from 8411.0 to 6214.0 U ml −1 min −1 as compared to Free Enzyme. Entrapped maltase displayed broad thermal stability up to 80°C whereas; Free Enzyme completely lost it activity when temperature reached up to 60°C. Scanning electron microscopy of agarose beads before and after maltase entrapment revealed significant morphological change on the matrix surface. Considering the economic feasibility, the entrapped maltase indicated imperative recycling efficiency up to ten reaction cycles.
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immobilization of pectin degrading Enzyme from bacillus licheniformis kibge ib 21 using agar agar as a support
Carbohydrate Polymers, 2014Co-Authors: Haneef Ur Rehman, Afsheen Aman, Raheela Rahmat Zohra, Shah Ali Ul QaderAbstract:Abstract Pectinase from Bacillus licheniformis KIBGE IB-21 was immobilized in agar-agar matrix using entrapment technique. Effect of different concentrations of agar-agar on pectinase immobilization was investigated and it was found that maximum immobilization was achieved at 3.0% agar-agar with 80% Enzyme activity. After immobilization, the optimum temperature of Enzyme increased from 45 to 50 °C and reaction time from 5 to 10 minutes as compared to Free Enzyme. Due to the limited diffusion of high molecular weight substrate, K m of immobilized Enzyme slightly increased from 1.017 to 1.055 mg ml −1 , while V max decreased from 23,800 to 19,392 μM min −1 as compared to Free Enzyme. After 120 h entrapped pectinase retained their activity up to 82% and 71% at 30 °C and 40 °C, respectively. The entrapped pectinase showed activity until 10th cycle and maintain 69.21% activity even after third cycle.
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degradation of complex carbohydrate immobilization of pectinase from bacillus licheniformis kibge ib21 using calcium alginate as a support
Food Chemistry, 2013Co-Authors: Haneef Ur Rehman, Afsheen Aman, Shah Ali Ul Qader, Antonio Molinaro, Alba Silipo, Asma AnsariAbstract:Abstract Pectinases are heterogeneous group of Enzymes that catalyse the hydrolysis of pectin substances which is responsible for the turbidity and undesirable cloudiness in fruits juices. In current study, partially purified pectinase from Bacillus licheniformis KIBGE-IB21 was immobilized in calcium alginate beads. The effect of sodium alginate and calcium chloride concentration on immobilization was studied and it was found that the optimal sodium alginate and calcium chloride concentration was 3.0% and 0.2 M, respectively. It was found that immobilization increases the optimal reaction time for pectin degradation from 5 to 10 min and temperature from 45 to 55 °C, whereas, the optimal pH remained same with reference to Free Enzyme. Thermal stability of Enzyme increased after immobilization and immobilized pectinase retained more than 80% of its initial activity after 5 days at 30 °C as compared with Free Enzyme which showed only 30% of residual activity. The immobilized Enzyme also exhibited good operational stability and 65% of its initial activity was observed during third cycle. In term of pectinase immobilization efficiency and stability, this calcium alginate beads approach seemed to permit good results and can be used to make a bioreactor for various applications in food industries.
Munish Puri - One of the best experts on this subject based on the ideXlab platform.
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Suitability of Recombinant Lipase Immobilised on Functionalised Magnetic Nanoparticles for Fish Oil Hydrolysis
MDPI AG, 2019Co-Authors: Madan L Verma, Nalam M Rao, Takuya Tsuzuki, Colin J Barrow, Munish PuriAbstract:Recombinant Bacillus subtilis lipase was immobilised on magnetic nanoparticles by a facile covalent method and applied to fish oil hydrolysis. High loading of Enzyme to the functionalised nanoparticle was achieved with a protein binding efficiency of 95%. Structural changes of the confined Enzyme on the surface of the nanoparticles was investigated using transmission electron microscopy and spectroscopic techniques (attenuated total reflectance-Fourier transform infrared and circular dichroism). The biocatalytic potential of immobilised lipase was compared with that of Free Enzyme and biochemically characterised with respect to different parameters such as pH, temperature, substrate concentrations and substrate specificity. The thermal stability of functionalised nanoparticle bound Enzyme was doubled that of Free Enzyme. Immobilised lipase retained more than 50% of its initial biocatalytic activity after recyclability for twenty cycles. The ability to the immobilised thermostable lipase to concentrate omega-3 fatty acids from fish oil was investigated. Using synthetic substrate, the immobilised Enzyme showed 1.5 times higher selectivity for docosahexaenoic acid (DHA), and retained the same degree of selectivity for eicosapentaenoic acid (EPA), when compared to the Free Enzyme
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Suitability of magnetic nanoparticle immobilised cellulases in enhancing enzymatic saccharification of pretreated hemp biomass
Biotechnology for Biofuels, 2014Co-Authors: Reinu E Abraham, Madan L Verma, Colin J Barrow, Munish PuriAbstract:Background Previous research focused on pretreatment of biomass, production of fermentable sugars and their consumption to produce ethanol. The main goal of the work was to economise the production process cost of fermentable sugars. Therefore, the objective of the present work was to investigate Enzyme hydrolysis of microcrystalline cellulose and hemp hurds (natural cellulosic substrate) using Free and immobilised Enzymes. Cellulase from Trichoderma reesei was immobilised on an activated magnetic support by covalent binding and its activity was compared with that of the Free Enzyme to hydrolyse microcrystalline cellulose and hemp hurds on the basis of thermostability and reusability. Results Up to 94% protein binding was achieved during immobilisation of cellulase on nanoparticles. Successful binding was confirmed using Fourier transform infrared spectroscopy (FTIR). The Free and immobilised Enzymes exhibited identical pH optima (pH 4.0) and differing temperature optima at 50°C and 60°C, respectively. The K_ M values obtained for the Free and immobilised Enzymes were 0.87 mg/mL and 2.6 mg/mL respectively. The immobilised Enzyme retained 50% Enzyme activity up to five cycles, with thermostability at 80°C superior to that of the Free Enzyme. Optimum hydrolysis of carboxymethyl cellulose (CMC) with Free and immobilised Enzymes was 88% and 81%, respectively. With pretreated hemp hurd biomass (HHB), the Free and immobilised Enzymes resulted in maximum hydrolysis in 48 h of 89% and 93%, respectively. Conclusion The current work demonstrated the advantages delivered by immobilised Enzymes by minimising the consumption of cellulase during substrate hydrolysis and making the production process of fermentable sugars economical and feasible. The activity of cellulase improved as a result of the immobilisation, which provided a better stability at higher temperatures. The immobilised Enzyme provided an advantage over the Free Enzyme through the reusability and longer storage stability properties that were gained as a result of the immobilisation.
