Improved stability and reusability of endoglucanase from Clostridium thermocellum by a biosilica-based auto-encapsulation method

Ryu, Young Ha; Yeo, Ki Baek; Ki, Mi-Ran; Kim, Yong Jun; Pack, Seung Pil

doi:10.1016/j.bej.2015.09.006

Cited by 23 publications

(10 citation statements)

References 24 publications

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“…Pellets were rinsed twice with PBS to remove unimmobilized B-E. The immobilization efficiency is calculated according to Pack et al, 48 using the following equation:…”

Section: Methodsmentioning

confidence: 99%

“…Increased thermal stabilities were also observed where enzymes were immobilized through biomimetic silicification. 48,64 These might be attributed to the enzymes encapsulation into biosilica matrix, which protects them from thermal denaturation. 48,64 In fact, the thermal stability of B-E@silica was still far from the demands of industrial application.…”

Section: Methodsmentioning

confidence: 99%

“…48,64 These might be attributed to the enzymes encapsulation into biosilica matrix, which protects them from thermal denaturation. 48,64 In fact, the thermal stability of B-E@silica was still far from the demands of industrial application. We previously reported that SpyTag/SpyCatcher-mediated cyclization of lichenase could significantly improve its thermal stability, even in boiling water.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Investigation of the ELP-Mediated Silicification-Based Protein Self-Immobilization Using an Acidic Target Enzyme

Lin

Qiu

Cai

et al. 2020

Ind. Eng. Chem. Res.

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Recently, we proposed a novel and effective strategy for enzyme immobilization, namely, as elastin-like polypeptide (ELP)-mediated silicification-based protein self-immobilization (ESPSI). Here, an enzyme with an acidic isoelectric point (pI) was chosen to further demonstrate the feasibility and generality of this method; pI is a critical factor in biosilicification. The lichenase (BglS) from Bacillus subtilis 168, with a theoretical pI of 5.77, was genetically fused to the cationic ELP. The recombinant chimera protein BglS-ELP (B-E) was purified through ELP-based nonchromatographic inverse transition cycling (ITC) method. B-E then was self-encapsulated within silica nanoparticle (NP) via ELP-mediated biomimetic silicification, producing the nanobiocatalyst B-E@silica. Excellent encapsulation efficiency (>85%) could be achieved within a short immobilization time (10 min). In addition, the encapsulation efficiency could be promoted through temperature-induced phase transition. Meanwhile, this method gave negligible protein leakage (<0.5%). Thus, the B-E@silica showed good reusability, retaining ∼80% of the initial activity after 10 reaction cycles. These results indicated that ESPSI was suitable for the protein with an acidic pI. In addition, phase transition was a unique route to improve immobilization efficiency.

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“…Pellets were rinsed twice with PBS to remove unimmobilized B-E. The immobilization efficiency is calculated according to Pack et al, 48 using the following equation:…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Investigation of the ELP-Mediated Silicification-Based Protein Self-Immobilization Using an Acidic Target Enzyme

Lin

Qiu

Cai

et al. 2020

Ind. Eng. Chem. Res.

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“…In addition, additives like polyallylamine (PAH or PAA), poly(amidoamine) dendrimers (PAMAM), and poly-L-lysine (PLL) can successfully immobilize enzymes, such as D-amino acid oxidase, glucose oxidase, horseradish peroxidase, and adenosine deaminase [49]. Despite the promising performances, there are few articles about the use of BIS as support for cellulase immobilisation in the literature [55].…”

Section: Introductionmentioning

confidence: 99%

Facile Cellulase Immobilisation on Bioinspired Silica

et al. 2022

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Cellulases are enzymes with great potential for converting biomass to biofuels for sustainable energy. However, their commercial use is limited by their costs and low reusability. Therefore, the scientific and industrial sectors are focusing on finding better strategies to reuse enzymes and improve their performance. In this work, cellulase from Aspergillus niger was immobilised through in situ entrapment and adsorption on bio-inspired silica (BIS) supports. To the best of our knowledge, this green effect strategy has never been applied for cellulase into BIS. In situ entrapment was performed during support synthesis, applying a one-pot approach at mild conditions (room temperature, pH 7, and water solvent), while adsorption was performed after support formation. The loading efficiency was investigated on different immobilisation systems by Bradford assay and FTIR. Bovine serum albumin (BSA) was chosen as a control to optimize cellulase loading. The residual activity of cellulase was analysed by the dinitro salicylic acid (DNS) method. Activity of 90% was observed for the entrapped enzyme, while activity of ~55% was observed for the adsorbed enzyme. Moreover, the supported enzyme systems were recycled five times to evaluate their reuse potential. The thermal and pH stability tests suggested that both entrapment and adsorption strategies can increase enzyme activity. The results highlight that the entrapment in BIS is a potentially useful strategy to easily immobilise enzymes, while preserving their stability and recycle potential.

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“…Many researchers have investigated the encapsulation of enzymes in silica in order to gain the benefits of immobilization, namely, high reusability and higher chemical and thermal stability [1][2][3][4][5][6][7][8][9][10][11]. Sol-gel-based technologies are especially promising, with their ease of synthesis and high loading capacity [9,12].…”

Section: Introductionmentioning

confidence: 99%

Enzyme Loading in Internally-Coated Capillary Tubes Via Kinetic Doping

Jensen

Yip

2020

Coatings

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Development of capillary tubes internally doped with enzymes is of great interest for microfluidic reactions, and kinetic doping could provide a facile, inexpensive method for their manufacture. Kinetic doping has previously been demonstrated to have a high loading capacity with thin films coated on flat-surface coverslips. Dip coating of these surfaces was developed with the eventual intention to coat different shapes and sizes of substrates. In this study, we expanded the use of kinetic doping to internally-coated capillary tubes. Parameters for internally doping capillary tubes were developed with rhodamine 6G, which produced internally-coated thin films with a 90 nm thickness. Horseradish peroxidase (HRP) was then loaded into the capillary tubes, with a 47,000× increase in concentration over the loading solution. After excluding surface-adsorbed protein, the increase in HRP concentration in the thin films over the loading solution was determined to be 9850×. The activity of the loaded HRP was determined to be 0.019 ± 0.003 U/mg and shown to have a stronger resistance to denaturation by methanol.

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Improved stability and reusability of endoglucanase from Clostridium thermocellum by a biosilica-based auto-encapsulation method

Cited by 23 publications

References 24 publications

Investigation of the ELP-Mediated Silicification-Based Protein Self-Immobilization Using an Acidic Target Enzyme

Investigation of the ELP-Mediated Silicification-Based Protein Self-Immobilization Using an Acidic Target Enzyme

Facile Cellulase Immobilisation on Bioinspired Silica

Enzyme Loading in Internally-Coated Capillary Tubes Via Kinetic Doping

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