In Situ Microscopy for In-line Monitoring of the Enzymatic Hydrolysis of Cellulose

Opitz, Britta; Prediger, Andreas; Lüder, Christian; Eckstein, Marrit; Hilterhaus, Lutz; Lindner, Patrick; Beutel, Sascha; Scheper, Thomas; Liese, Andreas

doi:10.1021/ac4008495

Cited by 12 publications

(4 citation statements)

References 20 publications

(26 reference statements)

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“…Just like in other fields of biotechnology (and chemical technology), observing the reaction progress through direct in situ measurements of substrate, product or both inside the reactor offers the general advantage of improved process monitoring and control [141][142][143][144][145][146][147][148]. In-operando applications require the use of transparent windows, optical fibers or immersed probes.…”

Section: In-operando Reaction Monitoring and Enzyme Activity Determinmentioning

confidence: 99%

Advanced characterization of immobilized enzymes as heterogeneous biocatalysts

2016

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Section: In-operando Reaction Monitoring and Enzyme Activity Determinmentioning

confidence: 99%

Advanced characterization of immobilized enzymes as heterogeneous biocatalysts

2016

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“…Biomass variability is another issue that pre-processing technologies can also address (Williams et al, 2016). However, such preprocessing technologies cannot be screened for performance in downstream operations with available analytical methods that are currently applicable for low concentration slurries alone (Hu et al, 2009;Goacher et al, 2012;Opitz et al, 2013;Malinowska et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

In Situ Rheological Method to Evaluate Feedstock Physical Properties Throughout Enzymatic Deconstruction

et al. 2018

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Feedstock physical properties determine not only downstream flow behavior, but also downstream process yields. Enzymatic treatment of pretreated feedstocks is greatly dependent on upstream feedstock physical properties and choice of pre-processing Technologies. Currently available enzyme assays have been developed to study biomass slurries at low concentrations of ≤1% w/w. At commercially relevant biomass concentrations of ≥15% w/w, pretreated feedstocks have sludge-like properties, where low free water restricts movement of unattached enzymes. This work is an account of the various steps taken to develop a method that helps identify the time needed for solid-like biomass slurries transition into liquid-like states during enzymatic hydrolysis. A pre-processing technology that enables feedstocks in achieving this transition sooner will greatly benefit enzyme kinetics and thereby overall process economics. Through this in situ rheological properties determining method, we compared a model feedstock, Avicel ® PH101 cellulose, with acid pretreated corn stover. Novozymes Cellic ® CTec2 (80 mg protein/g glucan) can reduce 25% (w/w) Avicel from solid-like to liquid-like state in 5.5 h, as the phase angles rise beyond 45 • at this time. The same slurry needed 5.3 h to achieve liquid-like state with Megazyme endoglucanase (40 mg protein/g glucan). After 10.8 h, CTec2 slurry reached a phase angle of 89 • or complete liquid-like state but Megazyme slurry peaked only to 64.7 • , possibly due to inhibition by cello-oligomers. Acid pretreated corn stover at 30% (w/w) with a CTec2 protein loading of 80 mg/g glucan exhibited a solid-like to liquid-like transition at 37.8 h, which reflects the combined inhibition of low water activity and presence of lignin. The acid pretreated slurry also never achieved complete liquid-like state due to the presence of biomass residue. This method is applicable in several scenarios comparing varying combinations of pre-processing technologies, feedstock types, pretreatment chemistries, and enzymes. Using this method, we can generate a process chain with optimal flow behavior at commercially-relevant conditions.

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“…Atomic force microscopy for topography has become widely used because it is a high resolution technique, but it only gives access to the sample surface [10]. Regarding dynamical information monitored during substrate catalysis, cellulose particle size changes can be followed in situ during enzyme hydrolysis [11], but the resolution is quite low and the technical setup is not straightforward. Overall, essential spatial and dynamical information can only be acquired separately.…”

Section: Introductionmentioning

confidence: 99%

Fluorescent Probes for Exploring Plant Cell Wall Deconstruction: A Review

Paës

2014

Molecules

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Plant biomass is a potential resource of chemicals, new materials and biofuels that could reduce our dependency on fossil carbon, thus decreasing the greenhouse effect. However, due to its chemical and structural complexity, plant biomass is recalcitrant to green biological transformation by enzymes, preventing the establishment of integrated bio-refineries. In order to gain more knowledge in the architecture of plant cell wall to facilitate their deconstruction, many fluorescent probes bearing various fluorophores have been devised and used successfully to reveal the changes in structural motifs during plant biomass deconstruction, and the molecular interactions between enzymes and plant cell wall polymers. Fluorescent probes are thus relevant tools to explore plant cell wall deconstruction.

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In Situ Microscopy for In-line Monitoring of the Enzymatic Hydrolysis of Cellulose

Cited by 12 publications

References 20 publications

Advanced characterization of immobilized enzymes as heterogeneous biocatalysts

Advanced characterization of immobilized enzymes as heterogeneous biocatalysts

In Situ Rheological Method to Evaluate Feedstock Physical Properties Throughout Enzymatic Deconstruction

Fluorescent Probes for Exploring Plant Cell Wall Deconstruction: A Review

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