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Lyophilization, also known as freeze-drying, is a widely used method for stabilization, improvement of long-term storage stability, and simplification of the handling of drugs and/or carrier systems. Lyophilization is time- and energy-consuming; hence, optimized processes are r ...

Cellulose is an important biopolymer primarily stored as plant cell wall material. Plant-synthesized cellulose forms elementary fibrils that are micrometers in length and 3–5 nm in dimensions. Cellulose is a dynamic structure, and its size and property vary in different cellulose-co ...

Common analytical challenges impact current work to estimate the cost of converting plant biomass to fermentable sugars. The most noteworthy are measuring cellulase and hemicellulase activities, cellulase and hemicellulase protein, biomass compositions (before and after ...

Although it has a deceptively simple primary structure, the collective organization of bulk cellulose, particularly as it exists in cellulose fibers in the cell walls of living plants and other organisms, is quite diverse and complex. While some experimental techniques, such as vibrati ...

Biomass exhibits structural and chemical complexity over multiple size scales, presenting many challenges to the effective characterization of these materials. The macroscopic nature of plants requires that some form of size reduction, such as dissection and microtomy, be perfo ...

Coherent Raman scattering (CRS) microscopy is a label-free method for chemical imaging, as it offers chemical specificity with orders of magnitude better sensitivity than the state-of-the-art confocal Raman scattering microscopy. Currently CRS technique includes coherent a ...

Flow cytometry (FCM) techniques have been developed for sorting mesophilic organisms, but the difficulty increases if the target microbes are thermophilic anaerobes. We demonstrate a reliable, high-throughput method of screening thermophilic anaerobic organisms using FCM ...

To efficiently deconstruct recalcitrant plant biomass to fermentable sugars in industrial processes, biocatalysts of higher performance and lower cost are required. The genetic diversity found in the metagenomes of natural microbial biomass decay communities may harbor such ...

Single-molecule fluorescence detection is an invaluable technique for the study of molecular behavior in biological systems, both in vitro and in vivo. In this chapter, we focus on detailed protocols that utilize Total Internal Reflection Fluorescence Microscopy (TIRF-M) to visual ...

Affinity electrophoresis is a simple and rapid tool for the analysis of protein-binding affinities to soluble polysaccharides. This approach is particularly suitable for the characterization of the carbohydrate-active enzymes that contain a carbohydrate-binding module a ...

Cadherin (CA) and cadherin-like (CADG) doublet domains from the complex polysaccharide-degrading marine bacterium, Saccharophagus degradans 2–40, demonstrated reversible calcium-dependent binding to different complex polysaccharides, which serve as growth subs ...

Experimental identification of carbohydrate-binding modules (CBM) and determination of ligand specificity of each CBM are complementary and compulsory steps for their characterization. Some CBMs are very specific for their primary substrate (e.g., cellulose), whereas othe ...

The substrate accessibility to enzyme has been considered as one of the most important factors for biomass conversion. To avoid the irreversible collapse of pore structure during the drying of sample, the measurement needs to be performed in a wet state. In this report, a thermoporometry method ...

Plant cell walls are dynamic structures that show changes in composition and configuration depending on the developmental stage, biotic, and abiotic factors. Therefore, it is necessary to have tools for visualizing the components of the cell wall in situ at any stage. Here, we describe how spec ...

The native complexity of plant cell walls makes research on them challenging. Hence, it is advantageous to have a diversity of tools that can be used to analyze and characterize plant cell walls. In this chapter, we describe one of two immunological approaches that can be employed for screening of pla ...

Plant cell walls are composed of three basic structural biomolecules: cellulose, hemicellulose, and lignin with cellulose being the most abundant biopolymer on earth. Cellulose is composed of cellodextrins, which are linear polymers of glucose, and considered to be microcrystall ...

Cell wall recalcitrance is the largest contributor to the high expense of lignocellulose conversion to biofuels (Himmel ME et al., Science 315:804–807, 2007). In response to this problem, researchers at the BioEnergy Science Center (BESC) are working to determine the contributing facto ...

Although a poor indicator of how a cellulase preparation will perform on biomass, the filter paper unit (FPU) still finds wide use in the literature as an apparent measure of performance efficacy. In actuality, the assessment of commercial enzyme preparation performance in terms of biomass c ...

There are two types of processive cellulases, exocellulases and processive endoglucanases. There are also two classes of exocellulases, ones that attack the reducing ends of cellulose chains and ones that attack the nonreducing ends. There are a number of ways of assaying processivity but ...

Heteroxylans are polysaccharides with a backbone composed of 1,4-linked β-d-xylosyl residues. In hardwoods some of these xylosyl residues are substituted at O-2 with 4-O-methyl α-d-glucuronic and occasionally with α-d-glucuronic acid. In grasses, the xylan backbone is predominan ...