Recent Advances in Enzyme Immobilization Utilizing Nanotechnology for Biocatalysis

T.sriwong, Kotchakorn; Matsuda, Tomoko

doi:10.1021/acs.oprd.1c00404

Cited by 40 publications

(19 citation statements)

References 156 publications

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“…To be effective and efficient in this process, the enzyme must be able to tolerate high temperatures and the presence of the residual organic solvent from cellulosic pretreatment. , Previous methods to improve cellulase for industrial use have included immobilization to polymer surfaces, , magnetic nanoparticles, − and biohybrid nanoparticles. , These strategies have resulted in increased cycles of reusability, storage stability, and enhancements under nonoptimal enzymatic conditions. However, our studies show that the impact of polymer conjugation under optimal conditions and immobilization has been shown to come at the sacrifice of some enzymatic activity upon immobilization. ,− …”

Section: Introductionmentioning

confidence: 99%

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

et al. 2022

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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,Ndimethyl 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.

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Section: Introductionmentioning

confidence: 99%

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

et al. 2022

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“…Immobilization of enzymes on solid surfaces has numerous advantages in industrial contexts, including reusability and enhanced catalytic stability. ,,,,− Strategies for preparation of immobilized enzymes (covalent tethering through both native and installed functional groups, carrier-free cross-linking, both specific and nonspecific adsorption, etc.) have been reviewed in-depth by several research groups. ,− Several published studies have explored PETase immobilization on metallic supports as a means of increasing stability by genetically fusing the enzyme to a metal-binding polyhistidine tag. , It has been shown that immobilization, even without introduction of spacer molecules to reduce steric hindrance, has the potential to enhance PETase activity and stability in harsh temperature and pH conditions , but the full scope of PETase immobilization has yet to be explored.…”

Section: Introductionmentioning

confidence: 99%

Directed Immobilization of PETase on Mesoporous Silica Enables Sustained Depolymerase Activity in Synthetic Wastewater Conditions

Zurier

Goddard

2022

ACS Appl. Bio Mater.

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Microplastic accumulation in terrestrial and aquatic environments is a growing environmental challenge. Biodegradation has shown promise as an intervention strategy for reducing the spread of microplastics. The wastewater treatment system is a key intervention point in microplastic biodegradation due to its pivotal role in the water cycle at the interface between human activity and the environmental. However, the best characterized microplastic degradation enzyme, PETase, lacks the stability to perform at scale in wastewater treatment. In this work, we show that genetic fusion of PETase to a silica binding peptide enables directed immobilization of the enzyme onto silica nanoparticles. PETase activity in simulated wastewater conditions is quantified by linear regression from time zero to the time of maximum fluorescence of a fluorescent oxidized product of PETase degradation of PET microfibers. Mesoporous silica is shown to be a superior support material to nonporous silica. The resulting biocatalytic nanomaterial has up to 2.5-fold enhanced stability and 6.2-fold increased activity compared to free enzyme in unbuffered, 40 °C simulated influent (ionic strength ∼15 mM). In unbuffered, 40 °C simulated effluent (ionic strength ∼700 μM), reaction velocity and overall catalytic activity were increased by the biocatalytic material 2.1-fold relative to free PETase. All reactions were performed in 0.2 mL volumes, and enzyme concentrations were normalized across both free and immobilized samples to 9 μg/mL. Site-directed mutagenesis is shown to be a complementary technique to directed immobilization, which may aid in optimization of the biomaterial for wastewater applications. PETase stabilization in application-relevant environments as shown here enables progress toward application of PETase for microplastic biodegradation in wastewater treatment.

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“…Nanotechnological interventions in biotechnology have forecasted the potential of “nanobiocatalysis” as a sustainable tool in food waste valorization. − The “waste-to-wealth” concept via an enzyme–nanomaterial system has been synchronized to produce industrially relevant biochemical products, viz. biodiesel, , bioactive ingredients, − and hydrolysates, , from several food residues.…”

Section: Introductionmentioning

confidence: 99%

Magnetic Halloysite Nanotube/α-Amylase Based Nanobiocatalytic Transformation of Food Processing Waste into an Active Fermentation Ingredient

Sillu

Agnihotri

Reddy

2022

ACS Sustainable Chem. Eng.

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The concept of “nanobiocatalysis” creates exciting opportunities for improving enzyme performance via immobilization onto nanomaterials. A nanobiocatalyst consisting of magnetic halloysite nanotube (MHNT)/α-amylase was evaluated to transform food processing waste into an active fermentation medium formulation for low-cost bioprocessing. A high loading of α-amylase (185.5 mg (g of support)−1) was achieved on the surface of MHNTs through polydopamine functionalization. We validated the establishment of an enzyme-support system retaining >89% catalytic activity (27332 IU (g of support)−1) with improved enzyme handling (>99.1% recovery) and reusability (>56% activity, 10 cycles). MHNTs remarkably improved the enzyme kinetics and thermodynamic characteristics along with operational and storage stabilities and mitigated the likely inhibitory effects of cellulose/metal ions as contaminants. In addition to facilitating a continuous production of reducing sugars from the extracted starch over 72 h, the nanobiocatalyst was equally effective in preparing a nutritive food waste hydrolysate as a fermentable medium substitute for batch culturing of E. coli and a single-cell protein (A. niger). The commercial relevance of waste hydrolysate was also investigated to promote calcite precipitation via Bacillus sp. induced biocementation. We evidenced that nanobiocatalyst-assisted “starch depolymerization” released more nutritional components into the hydrolysate suspension, easily accessible to growing microbial cultures.

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Recent Advances in Enzyme Immobilization Utilizing Nanotechnology for Biocatalysis

Cited by 40 publications

References 156 publications

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

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

Directed Immobilization of PETase on Mesoporous Silica Enables Sustained Depolymerase Activity in Synthetic Wastewater Conditions

Magnetic Halloysite Nanotube/α-Amylase Based Nanobiocatalytic Transformation of Food Processing Waste into an Active Fermentation Ingredient

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