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.

admission. This proportion could already be greater in some parts of the country and may increase if referrals of cases of self-poisoning increase faster than the facilities for their assessment and management. The provision of social work and psychiatric expertise in casualty departments may be one means of preventing unnecessary medical admissions without risk to the patients. Dr Blake's and Dr Bramble's figures do not demonstrate, however, that any advantage would attach to medical teams taking over assessment from psychiatrists except that, by implication, assessments would be completed sooner by staff working on the ward full time. What the figures actually suggest is that if assessment of overdoses were left to house doctors there would be an increase in admissions to psychiatric units (by 19°U), outpatients (by 5O°'), and referrals to social services (by 140o). So for house doctors to assess overdoses would provide no economy for the psychiatric or social services. The study does not tell us what the consequences would have been for the six patients who the psychiatrists would have admitted but to whom the house doctors would have offered outpatient appointments. E J SALTER

Vitamin D deficiency.

This paper reports the first phase of a research program on visual perception of motion patterns characteristic of living organisms in locomotion. Such motion patterns in animals and men are termed here as biological motion. They are characterized by a far higher degree of complexity than the patterns of simple mechanical motions usually studied in our laboratories. In everyday perceptions, the visual information from biological motion and from the corresponding figurative contour patterns (the shape of the body) are intermingled. A method for studying information from the motion pattern per se without interference with the form aspect was devised. In short, the motion of the living body was represented by a few bright spots describing the motions of the main joints. It is found that 10–12 such elements in adequate motion combinations in proximal stimulus evoke a compelling impression of human walking, running, dancing, etc. The kinetic-geometric model for visual vector analysis originally developed in the study of perception of motion combinations of the mechanical type was applied to these biological motion patterns. The validity of this model in the present context was experimentally tested and the results turned out to be highly positive.

/pdf/visual-perception-of-biological-motion-and-a-model-for-its-4vc9i7xan0.pdf

Visual perception of biological motion and a model for its analysis

Lignocellulosic biomass has long been recognized as a potential sustainable source of mixed sugars for fermentation to biofuels and other biomaterials. Several technologies have been developed during the past 80 years that allow this conversion process to occur, and the clear objective now is to make this process cost-competitive in today's markets. Here, we consider the natural resistance of plant cell walls to microbial and enzymatic deconstruction, collectively known as "biomass recalcitrance." It is this property of plants that is largely responsible for the high cost of lignocellulose conversion. To achieve sustainable energy production, it will be necessary to overcome the chemical and structural properties that have evolved in biomass to prevent its disassembly.

http://science.sciencemag.org/content/sci/315/5813/804.full.pdf

Biomass recalcitrance: engineering plants and enzymes for biofuels production.

The amino acid sequences of 301 glycosyl hydrolases and related enzymes have been compared. A total of 291 sequences corresponding to 39 EC entries could be classified into 35 families. Only ten sequences (less than 5% of the sample) could not be assigned to any family. With the sequences available for this analysis, 18 families were found to be monospecific (containing only one EC number) and 17 were found to be polyspecific (containing at least two EC numbers). Implications on the folding characteristics and mechanism of action of these enzymes and on the evolution of carbohydrate metabolism are discussed. With the steady increase in sequence and structural data, it is suggested that the enzyme classification system should perhaps be revised.

A classification of glycosyl hydrolases based on amino acid sequence similarities.

A critical review of cellobiose dehydrogenases.

Perceptual organization during short tachistoscopic presentation of stimulus patterns formed by ten moving bright spots, representing a human body in walking, running, etc. was investigated. Exposure times were .1 sec to .5 sec. The results reveal that in all Ss the dot pattern is perceptually organized to a “gestalt”, a walking, running, etc., person at an exposure time of .2 sec. 40% of Ss perceived a human body in such motion at presentation times as short as 0.1 sec. Under the experimental conditions used the track length of the bright spots at the threshold of integration to a moving unit was of the size order 10′ visual angle. This result is regarded as indicating that a complex vector analysis of the proximal motion pattern is accomplished at the initial stage of physiological signal recording and that it is a consequence of receptive field organization. It is discussed in terms of vector calculus.

Spatio-temporal differentiation and integration in visual motion perception. An experimental and theoretical analysis of calculus-like functions in visual data processing.

EGIII, a new endoglucanase from Trichoderma reesei: the characterization of both gene and enzyme

The object of this investigation was to determine the distribution of brake reaction times which can be expected from drivers who have to brake suddenly and completely unexpectedly in traffic situations. The experiments were carried out as follows:1.Brake reaction time was measured on a large group of drivers (321), in an anticipated situation on the road (Brake reaction time 1).2.A small group of drivers (5) was repeatedly tested in the same way (Brake reaction time 2).3.The same small group was repeatedly tested in a surprise situation (Brake reaction time 3).4.The ratio of brake reaction time 3 to brake reaction time 2 was used as a correction factor and applied to brake reaction time 1.The corrected median of the resulting distribution was 0.9 sec.; 25% of the group was estimated to have a brake reaction time longer than 1.2 sec.

Drivers' Brake Reaction Times:

The inhibition effect of cellobiose on the initial stage of hydrolysis when cellobiohydrolase Cel 7A and endoglucanases Cel 7B, Cel 5A, and Cel 12A from Trichoderma reesei were acting on bacterial cellulose and amorphous cellulose that were [(3)H]- labeled at the reducing end was quantified. The apparent competitive inhibition constant (K(i)) for Cel 7A on [(3)H]-bacterial cellulose was found to be 1.6 +/- 0.5 mM, 100-fold higher than that for Cel 7A acting on low-molecular-weight model substrates. The hydrolysis of [(3)H]-amorphous cellulose by endoglucanases was even less affected by cellobiose inhibition with apparent K(i) values of 11 +/- 3 mM and 34 +/- 6 mM for Cel 7B and Cel 5A, respectively. Contrary to the case for the other enzymes studied, the release of radioactive label by Cel 12A was stimulated by cellobiose, possibly due to a more pronounced transglycosylating activity. Theoretical analysis of the inhibition of Cel 7A by cellobiose predicted an inhibition analogous to that of mixed type with two limiting cases, competitive inhibition if the prevalent enzyme-substrate complex without inhibitor is productive and conventional mixed type when the prevalent enzyme-substrate complex is nonproductive.

Gunnar Johansson

Papers

A critical review of cellobiose dehydrogenases.

Spatio-temporal differentiation and integration in visual motion perception. An experimental and theoretical analysis of calculus-like functions in visual data processing.

EGIII, a new endoglucanase from Trichoderma reesei: the characterization of both gene and enzyme

Drivers' Brake Reaction Times:

Inhibition of the Trichoderma reesei cellulases by cellobiose is strongly dependent on the nature of the substrate.