Modeling Progression of Fluorescent Probes in Bioinspired Lignocellulosic Assemblies

Paës, Gabriel; Burr, Sally; Saab, Marie-Belle; Molinari, Michaël; Aguié‐Béghin, Véronique; Chabbert, Brigitte

doi:10.1021/bm400338b

Cited by 13 publications

(23 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Indeed, such probes are considered as having few or no interaction with plant cell wall polymers [35, 36] and their size and low dispersity (Table 2) are in the range of typical enzymes degrading plant cell walls like cellulases and xylanases [38, 54, 55]. Rhodamine B was chosen as the fluorophore appended to the dextran probe since rhodamine B is excited beyond 540 nm, which is quite far from the maximum lignin excitation that is around 350–400 nm [23].…”

Section: Resultsmentioning

confidence: 99%

“…For FRAP experiments, prebleaching was performed with laser at 20% of its power and acquisition of five reference images, followed by bleaching with laser at 100%, with acquisition of 40 images and finally post-bleaching (laser at 20%) with acquisition of images until the bleached ROI intensity was constant. Calculation of the diffusion D and the mobile fraction MF were performed as previously described [35, 36]. …”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Exploring accessibility of pretreated poplar cell walls by measuring dynamics of fluorescent probes

Paës

Habrant

Ossemond

et al. 2017

Biotechnol Biofuels

Self Cite

View full text Add to dashboard Cite

BackgroundThe lignocellulosic cell wall network is resistant to enzymatic degradation due to the complex chemical and structural features. Pretreatments are thus commonly used to overcome natural recalcitrance of lignocellulose. Characterization of their impact on architecture requires combinatory approaches. However, the accessibility of the lignocellulosic cell walls still needs further insights to provide relevant information.ResultsPoplar specimens were pretreated using different conditions. Chemical, spectral, microscopic and immunolabeling analysis revealed that poplar cell walls were more altered by sodium chlorite-acetic acid and hydrothermal pretreatments but weakly modified by soaking in aqueous ammonium. In order to evaluate the accessibility of the pretreated poplar samples, two fluorescent probes (rhodamine B-isothiocyanate–dextrans of 20 and 70 kDa) were selected, and their mobility was measured by using the fluorescence recovery after photobleaching (FRAP) technique in a full factorial experiment. The mobility of the probes was dependent on the pretreatment type, the cell wall localization (secondary cell wall and cell corner middle lamella) and the probe size. Overall, combinatory analysis of pretreated poplar samples showed that even the partial removal of hemicellulose contributed to facilitate the accessibility to the fluorescent probes. On the contrary, nearly complete removal of lignin was detrimental to accessibility due to the possible cellulose–hemicellulose collapse.ConclusionsEvaluation of plant cell wall accessibility through FRAP measurement brings further insights into the impact of physicochemical pretreatments on lignocellulosic samples in combination with chemical and histochemical analysis. This technique thus represents a relevant approach to better understand the effect of pretreatments on lignocellulose architecture, while considering different limitations as non-specific interactions and enzyme efficiency.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Exploring accessibility of pretreated poplar cell walls by measuring dynamics of fluorescent probes

Paës

Habrant

Ossemond

et al. 2017

Biotechnol Biofuels

Self Cite

View full text Add to dashboard Cite

show abstract

“…Mathematically speaking, the response is equal to a sum of parameters and their combination, each having a coefficient related to their importance on the response. So if the parameters computed have numerical levels, the equation can be used to predict the response in conditions not tested experimentally [10]. The values for each parameter must remain between the highest and lowest levels used; otherwise, the prediction is not statistically acceptable.…”

Section: Statistical Analysis To Highlight Meaningful Parametersmentioning

confidence: 99%

“…These bioinspired assemblies are based on a hemicellulose network whose polymers (feruloylated arabinoxylans, FAXs) are cross-linked through ferulic acid moieties to make gels under the action of a peroxidase [8,9]. This network can be complexified by adding cellulose nano-crystals (CNCs) [10,11]. Type and concentration of both polymers can be easily varied.…”

Section: Introductionmentioning

confidence: 99%

Bioinspired Assemblies of Plant Cell Walls for Measuring Protein-Carbohydrate Interactions by FRAP

Paës

2017

Methods in Molecular Biology

Self Cite

View full text Add to dashboard Cite

The interactions of proteins involved in plant cell wall hydrolysis, such as enzymes and CBMs, significantly determine their role and efficiency. In order to go beyond the characterization of interactions with simple ligands, bioinspired assemblies combined with the measurement of diffusion and interaction by FRAP offer a relevant alternative for highlighting the importance of different parameters related to the protein affinity and to the assembly.

show abstract

“…Consequently, understanding and modelling the relationship between accessibility and hydrolysis is critical. 12,13 Regarding enzymes, some of them can be appended to one or several non-enzymatic modules. 14,15 Some of these modules are carbohydrate-binding modules (CBMs), defined as ''a contiguous amino acid sequence within a carbohydrate-active enzyme with a discrete fold having carbohydrate-binding activity''.…”

Section: Introductionmentioning

confidence: 99%

Bioinspired assemblies of plant cell wall polymers unravel the affinity properties of carbohydrate-binding modules

Paës¹,

Schantz²,

Ohlin³

2015

Soft Matter

Self Cite

View full text Add to dashboard Cite

Lignocellulose-acting enzymes play a central role in the biorefinery of plant biomass to make fuels, chemicals and materials. These enzymes are often appended to carbohydrate binding modules (CBMs) that promote substrate targeting. When used in plant materials, which are complex assemblies of polymers, the binding properties of CBMs can be difficult to understand and predict, thus limiting the efficiency of enzymes. In order to gain more information on the binding properties of CBMs, some bioinspired model assemblies that contain some of the polymers and covalent interactions found in the plant cell walls have been designed. The mobility of three engineered CBMs has been investigated by FRAP in these assemblies, while varying the parameters related to the polymer concentration, the physical state of assemblies and the oligomerization state of CBMs. The features controlling the mobility of the CBMs in the assemblies have been quantified and hierarchized. We demonstrate that the parameters can have additional or opposite effects on mobility, depending on the CBM tested. We also find evidence of a relationship between the mobility of CBMs and their binding strength. Overall, bioinspired assemblies are able to reveal the unique features of affinity of CBMs. In particular, the results show that oligomerization of CBMs and the presence of ferulic acid motifs in the assemblies play an important role in the binding affinity of CBMs. Thus we propose that these features should be finely tuned when CBMs are used in plant cell walls to optimise bioprocesses.

show abstract

Modeling Progression of Fluorescent Probes in Bioinspired Lignocellulosic Assemblies

Cited by 13 publications

References 38 publications

Exploring accessibility of pretreated poplar cell walls by measuring dynamics of fluorescent probes

Exploring accessibility of pretreated poplar cell walls by measuring dynamics of fluorescent probes

Bioinspired Assemblies of Plant Cell Walls for Measuring Protein-Carbohydrate Interactions by FRAP

Bioinspired assemblies of plant cell wall polymers unravel the affinity properties of carbohydrate-binding modules

Contact Info

Product

Resources

About