Bioseparation of recombinant cellulose-binding module-proteins by affinity adsorption on an ultra-high-capacity cellulosic adsorbent

Hong, Jiong; Ye, Xinhao; Zhang, Y.‐H. Percival

doi:10.1016/j.aca.2008.05.041

Cited by 94 publications

(87 citation statements)

References 40 publications

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“…Furthermore, the binding capacity of enzymes to RAC was 10 times higher than that to Avicel, which was similar to that reported by Hong et al [34]. The E max of wild-type and mutant AcCel12B to Avicel and RAC were not different from those of other cellulases [34,35]. Srisodsuk et al previously noted that the linker deletions of CBH I from Trichoderma reesei did not affect the enzyme affinity on crystalline cellulose at an initial enzyme concentration of 0.2 µM (5-9% saturation), but decreased the binding capacity as the enzyme concentration approached 10 µM [6].…”

Section: Adsorption and Desorption Of Wild-type And Mutant Accel12bsupporting

confidence: 90%

“…The E max (maximum adsorption capacities) of wild-type and mutant AcCel12B were not significantly different. Furthermore, the binding capacity of enzymes to RAC was 10 times higher than that to Avicel, which was similar to that reported by Hong et al [34]. The E max of wild-type and mutant AcCel12B to Avicel and RAC were not different from those of other cellulases [34,35].…”

Section: Adsorption and Desorption Of Wild-type And Mutant Accel12bsupporting

confidence: 89%

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The PT/S-Box of Modular Cellulase AcCel12B Plays a Key Role in the Hydrolysis of Insoluble Cellulose

Wang

et al. 2018

Catalysts

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Abstract:Cellulases play key roles in the degradation of lignocellulosic materials. The function and mechanism of the catalytic domain (CD) and carbohydrate-binding module (CBM) of cellulases were earlier revealed by analysis and characterization of protein structure. However, understanding of the catalytic mechanism of the entire enzyme, and the analysis of the catalytic model, were inadequate. Therefore, the linker chain between CD and CBM has been extensively studied to bridge this gap. Cellulase AcCel12B and three mutants with different linker lengths (with no or 1-3 PT/S-box units) were successfully constructed and purified. Results showed that the activity of cellulases on Avicel and regenerated amorphous cellulose (RAC) increased with the number of PT/S-box units. Furthermore, the desorption of AcCel12B and its mutants from RAC and Avicel were significantly different. The energy of desorption of wild-type and mutant AcCel12B from cellulose decreased with the number of PT/S-box units. Thus, AcCel12B containing more PT/S-box units was more easily desorbed and had more opportunity to hydrolyze cellulose than other samples. The number of PT/S-box units in endocellulase affected the desorption of the enzyme, which is possibly responsible for the differences in the activity of wild-type and mutant AcCel12B on Avicel and RAC.

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Section: Adsorption and Desorption Of Wild-type And Mutant Accel12bsupporting

confidence: 90%

Section: Adsorption and Desorption Of Wild-type And Mutant Accel12bsupporting

confidence: 89%

The PT/S-Box of Modular Cellulase AcCel12B Plays a Key Role in the Hydrolysis of Insoluble Cellulose

Wang

et al. 2018

Catalysts

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“…The tub-ground materials were approximately 9 months old (harvested in fall 2005). The recombinant thioredoxin-green fluorescent protein-cellulose binding module (TGC) fusion protein was produced in Escherichia coli BL21 (pNT02) (Hong et al, 2007) and purified by affinity adsorption on RAC followed by modest desorption using ethyl glycol (EG) (Hong et al, 2008b). The EG was removed by using membrane dialysis in a 50 mM sodium citrate buffer (pH 6.0).…”

Section: Chemicals and Microorganismmentioning

confidence: 99%

“…This new approach more accurately assesses substrate characteristic related to enzymatic cellulose hydrolysis than traditional methods such as nitrogen adsorption-based Brunauer-EmmettTeller (BET), size exclusion, and small angle X-ray scattering (Hong et al, 2007;Zhang and Lynd, 2004). Regenerated amorphous cellulose (RAC) that is prepared from microcrystalline cellulose (Avicel) has $20-fold higher CAC (Hong et al, 2007(Hong et al, , 2008b and exhibits much faster enzymatic hydrolysis rates than microcrystalline cellulose (Zhang et al, 2006a), which is in agreement with the model prediction that increasing CAC is more important for increasing hydrolysis rates than decreasing DP (Zhang and Lynd, 2006). The CAC value of Avicel (m 2 per gram of Avicel) based on the TGC adsorption was only one-tenth of that based on the BET method (Marshall and Sixsmith, 1974), implying that about 90% gross surface area measure based on nitrogen adsorption cannot be accessible to largesize cellulase protein molecules, at least initially.…”

Section: Introductionmentioning

confidence: 99%

Comparative study of corn stover pretreated by dilute acid and cellulose solvent‐based lignocellulose fractionation: Enzymatic hydrolysis, supramolecular structure, and substrate accessibility

Zhu

Sathitsuksanoh

Vinzant

et al. 2009

Biotech & Bioengineering

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200

164

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Liberation of fermentable sugars from recalcitrant biomass is among the most costly steps for emerging cellulosic ethanol production. Here we compared two pretreatment methods (dilute acid, DA, and cellulose solvent and organic solvent lignocellulose fractionation, COSLIF) for corn stover. At a high cellulase loading [15 filter paper units (FPUs) or 12.3 mg cellulase per gram of glucan], glucan digestibilities of the corn stover pretreated by DA and COSLIF were 84% at hour 72 and 97% at hour 24, respectively. At a low cellulase loading (5 FPUs per gram of glucan), digestibility remained as high as 93% at hour 24 for the COSLIF-pretreated corn stover but reached only $60% for the DA-pretreated biomass. Quantitative determinations of total substrate accessibility to cellulase (TSAC), cellulose accessibility to cellulase (CAC), and non-cellulose accessibility to cellulase (NCAC) based on adsorption of a nonhydrolytic recombinant protein TGC were measured for the first time. The COSLIF-pretreated corn stover had a CAC of 11.57 m 2 /g, nearly twice that of the DA-pretreated biomass (5.89 m 2 /g). These results, along with scanning electron microscopy images showing dramatic structural differences between the DA-and COSLIF-pretreated samples, suggest that COSLIF treatment disrupts microfibrillar structures within biomass while DA treatment mainly removes hemicellulose. Under the tested conditions COSLIF treatment breaks down lignocellulose structure more extensively than DA treatment, producing a more enzymatically reactive material with a higher CAC accompanied by faster hydrolysis rates and higher enzymatic digestibility.

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“…In reality, several low-cost scalable protein purification approaches are available, for example, simple centrifugation for secretory enzymes, adsorption/desorption on low-cost cellulosic materials [74,75] (Figure 5), heat precipitation for thermostable enzymes [65,76] (Figure 6), and ammonia precipitation [77] (Figure 6). Therefore, the purification costs for bulk enzymes would become relatively minor.…”

Section: Sypab Challenges and Opportunitiesmentioning

confidence: 99%

Renewable Hydrogen Carrier — Carbohydrate: Constructing the Carbon-Neutral Carbohydrate Economy

Zhang

Mielenz

2011

Energies

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Abstract:The hydrogen economy presents an appealing energy future but its implementation must solve numerous problems ranging from low-cost sustainable production, high-density storage, costly infrastructure, to eliminating safety concern. The use of renewable carbohydrate as a high-density hydrogen carrier and energy source for hydrogen production is possible due to emerging cell-free synthetic biology technology-cell-free synthetic pathway biotransformation (SyPaB

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Bioseparation of recombinant cellulose-binding module-proteins by affinity adsorption on an ultra-high-capacity cellulosic adsorbent

Cited by 94 publications

References 40 publications

The PT/S-Box of Modular Cellulase AcCel12B Plays a Key Role in the Hydrolysis of Insoluble Cellulose

The PT/S-Box of Modular Cellulase AcCel12B Plays a Key Role in the Hydrolysis of Insoluble Cellulose

Comparative study of corn stover pretreated by dilute acid and cellulose solvent‐based lignocellulose fractionation: Enzymatic hydrolysis, supramolecular structure, and substrate accessibility

Renewable Hydrogen Carrier — Carbohydrate: Constructing the Carbon-Neutral Carbohydrate Economy

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