Cellulases Dig Deep

Bubner, Patricia; Dohr, Judith; Plank, Harald; Mayrhofer, Claudia; Nidetzky, Bernd

doi:10.1074/jbc.m111.257717

Cited by 55 publications

(31 citation statements)

References 33 publications

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“…The complete cellulase system of Trichoderma reesei mutant SVG 17 was produced and stored as previously described (7). Pure preparations of β-glucosidase (BGL) from Aspergillus niger and CBH I and EG, both from T. longibrachiatum , were purchased from Megazyme (Dublin, Ireland).…”

Section: Methodsmentioning

confidence: 99%

“…The cellulosic substrate employed in this study was prepared and characterized as previously described (7). The protocol involves dissolution of the microcrystalline cellulose Avicel (Sigma-Aldrich) in ionic liquid.…”

Section: Methodsmentioning

confidence: 99%

“…The resulting transparent gel was subjected to a fractional dehydration with ethanol (30% to absolute). This procedure removed the residual ionic liquid, leaving a pure and polymorphic cellulosic substrate (7). …”

Section: Methodsmentioning

confidence: 99%

“…The actual source of this limitation, however, be it the enzymes, the substrate, or both, is a fundamentally unsolved puzzle (14). Identifying the limiting factors by achieving a deeper mechanistic understanding of enzymatic cellulose degradation is pivotal for advancing biomass-to-fuel process development (5–7). …”

Section: Introductionmentioning

confidence: 99%

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Dissecting and Reconstructing Synergism

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Bubner

Eibinger

et al. 2012

Journal of Biological Chemistry

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Background: Synergistic interplay of cellulases is key for efficiency of cellulose hydrolysis.Results: In situ observation of individual and synergistic action of endo- and exo-cellulases on a polymorphic cellulose substrate reveals specificity of individual enzyme components for crystalline or amorphous regions.Conclusion: Cellulase synergism is governed by mesoscopic morphological characteristics of the cellulose substrate.Significance: Advanced knowledge basis for rational optimization of cellulose saccharification.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Dissecting and Reconstructing Synergism

Ganner

Bubner

Eibinger

et al. 2012

Journal of Biological Chemistry

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“…[65] Time-lapse atomic force microscopy (AFM) imaging has also been used to correlate substrate digestability with topological features. [66] Optimization of cellulosomes requires the establishment of reference assays to quantify the efficiency of the different enzymes and their mutants. [63] Several factors complicate the achievement of this goal: Figure 6.…”

Section: Bacterial Breakdown Of Crystalline Cellulosementioning

confidence: 99%

Nanoscale Engineering of Designer Cellulosomes

et al. 2016

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Biocatalysts showcase the upper limit obtainable for high-speed molecular processing and transformation. Efforts to engineer functionality in synthetic nanostructured materials are guided by the increasing knowledge of evolving architectures, which enable controlled molecular motion and precise molecular recognition. The cellulosome is a biological nanomachine, which, as a fundamental component of the plant-digestion machinery from bacterial cells, has a key potential role in the successful development of environmentally-friendly processes to produce biofuels and fine chemicals from the breakdown of biomass waste. Here, the progress toward so-called "designer cellulosomes", which provide an elegant alternative to enzyme cocktails for lignocellulose breakdown, is reviewed. Particular attention is paid to rational design via computational modeling coupled with nanoscale characterization and engineering tools. Remaining challenges and potential routes to industrial application are put forward.

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Visualizing cellulase activity

Bubner

Plank

Nidetzky

2013

Biotech & Bioengineering

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Commercial exploitation of lignocellulose for biotechnological production of fuels and commodity chemicals requires efficient-usually enzymatic-saccharification of the highly recalcitrant insoluble substrate. A key characteristic of cellulose conversion is that the actual hydrolysis of the polysaccharide chains is intrinsically entangled with physical disruption of substrate morphology and structure. This "substrate deconstruction" by cellulase activity is a slow, yet markedly dynamic process that occurs at different length scales from and above the nanometer range. Little is currently known about the role of progressive substrate deconstruction on hydrolysis efficiency. Application of advanced visualization techniques to the characterization of enzymatic degradation of different celluloses has provided important new insights, at the requisite nano-scale resolution and down to the level of single enzyme molecules, into cellulase activity on the cellulose surface. Using true in situ imaging, dynamic features of enzyme action and substrate deconstruction were portrayed at different morphological levels of the cellulose, thus providing new suggestions and interpretations of rate-determining factors. Here, we review the milestones achieved through visualization, the methods which significantly promoted the field, compare suitable (model) substrates, and identify limiting factors, challenges and future tasks.

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Cellulases Dig Deep

Cited by 55 publications

References 33 publications

Dissecting and Reconstructing Synergism

Dissecting and Reconstructing Synergism

Nanoscale Engineering of Designer Cellulosomes

Visualizing cellulase activity

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