Fundamental features of microbial cellulose utilization are examined at successively higher levels of aggregation encompassing the structure and composition of cellulosic biomass, taxonomic diversity, cellulase enzyme systems, molecular biology of cellulase enzymes, physiology of cellulolytic microorganisms, ecological aspects of cellulase-degrading communities, and rate-limiting factors in nature. The methodological basis for studying microbial cellulose utilization is considered relative to quantification of cells and enzymes in the presence of solid substrates as well as apparatus and analysis for cellulose-grown continuous cultures. Quantitative description of cellulose hydrolysis is addressed with respect to adsorption of cellulase enzymes, rates of enzymatic hydrolysis, bioenergetics of microbial cellulose utilization, kinetics of microbial cellulose utilization, and contrasting features compared to soluble substrate kinetics. A biological perspective on processing cellulosic biomass is presented, including features of pretreated substrates and alternative process configurations. Organism development is considered for "consolidated bioprocessing" (CBP), in which the production of cellulolytic enzymes, hydrolysis of biomass, and fermentation of resulting sugars to desired products occur in one step. Two organism development strategies for CBP are examined: (i) improve product yield and tolerance in microorganisms able to utilize cellulose, or (ii) express a heterologous system for cellulose hydrolysis and utilization in microorganisms that exhibit high product yield and tolerance. A concluding discussion identifies unresolved issues pertaining to microbial cellulose utilization, suggests approaches by which such issues might be resolved, and contrasts a microbially oriented cellulose hydrolysis paradigm to the more conventional enzymatically oriented paradigm in both fundamental and applied contexts.

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Microbial cellulose utilization: fundamentals and biotechnology.

Bacterial biofilms in nature and disease.

Publisher Summary The chapter focuses on the recent advances in understanding the structural and functional organization of individual cellulases, their regulation, and the ways in which the multiple enzyme components of cellulolytic systems cooperate. It overviews the cellulose structures because cellulose is more than a homopolymer of β-1 ,4 linked glucose units. An appreciation of its complex physical organization and its interactions with other plant cell wall components is central for understanding of the mechanisms of cellulase action. Cellulose nearly always occurs in close association with plant cell wall matrix polysaccharides so that enzymes such as xylanases are intimately involved in the attack of cellulose in vivo. Cellulases effect important changes to their substrate before releasing soluble products and the key to understanding cellulase action rest in the examination of these events. Progress in this area is limited by the availability of appropriate analytical tools although new techniques, such as atomic force microscopy are promising. The properties of cellulases are profoundly altered by the presence of trace enzyme contaminants. Future studies in vitro are proposed to be restricted to enzymes from recombinant sources. The reasons for the individual cellulolytic bacteria and fungi requiring many related cellulases with specificities that overlap is still not clear, but perhaps this is because the complexity of the substrates and of the task these microorganisms face is underestimated.

Cellulose hydrolysis by bacteria and fungi

Direct microscopic examination of the rumen and its contents shows microbial populations largely attached to feed particles in the digesta Most feeds contain a surface layer that is resistant to attachment and therefore to digestion Infiltration of these recalcitrant epidermal layers through damage sites or through focused enzymatic attack is essential for initiation of the digestive process Proliferation of primary colonizing cells produces glycocalyx-enclosed microcolonies Secondary colonizers from the ruminal fluid associate with microcolonies, resulting in the formation of multispecies microbial biofilms These metabolically related organisms associate with their preferred substrates and produce the myriad of enzymes necessary for the digestion of chemically and structurally complex plant tissues Upon accessing the internal, enzyme-susceptible tissues, microbial "digestive consortia" attach to a variety of nutrients, including protein, cellulose, and starch and digest insoluble feed materials from the inside out Substances that prevent microbial attachment or promote detachment (eg, condensed tannins, methylcellulose) can completely inhibit cellulose digestion As the microbial consortium matures and adapts to a particular type of feed, it becomes inherently stable and its participant microorganisms are notoriously difficult to manipulate due to the impenetrable nature of biofilms Properties of feed that place constraints on microbial attachment and biofilm formation can have a profound effect on both the rate and extent of feed digestion in the rumen Developments in feed processing (ie, chemical and physical), plant breeding, and genetic engineering (both of ruminal microorganisms and plants) that overcome these constraints through the promotion of microbial attachment and biofilm formation could substantially benefit ruminant production

Microbial attachment and feed digestion in the rumen.

In anaerobic environments rich in decaying plant material, the decomposition of cellulose is brought about by complex communities of interacting microorganisms. Because the substrate, cellulose, is insoluble, bacterial and fungal degradation occurs exocellularly, either in association with the outer cell envelope layer or extracellularly. Products of cellulose hydrolysis are available as carbon and energy sources for other microbes that inhabit environments in which cellulose is biodegraded, and this availability forms the basis of many microbial interactions that occur in these environments. This review discusses interactions among members of cellulose-decomposing microbial communities in various environments. It considers cellulose decomposing communities in soils, sediments, and aquatic environments, as well as those that degrade cellulose in association with animals. These microbial communities contribute significantly to the cycling of carbon on a global scale.

Cellulose Degradation in Anaerobic Environments

The presence of methylellulose prevents the attachment of cellulolytic rumen bacteria to cellulose fibers. The addition of methylcellulose to pure cultures of these organisms in which the cells are...

K.-J. Cheng

Papers

Electron microscopic study of the methylcellulose-mediated detachment of cellulolytic rumen bacteria from cellulose fibers.