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.

Bioconversion of forest products industry waste cellulosics to fuel ethanol: a review.

Biological conversion of cellulosic biomass to fuels and chemicals offers the high yields to products vital to economic success and the potential for very low costs. Enzymatic hydrolysis that converts lignocellulosic biomass to fermentable sugars may be the most complex step in this process due to substrate-related and enzyme-related effects and their interactions. Although enzymatic hydrolysis offers the potential for higher yields, higher selectivity, lower energy costs and milder operating conditions than chemical processes, the mechanism of enzymatic hydrolysis and the relationship between the substrate structure and function of various glycosyl hydrolase components is not well understood. Consequently, limited success has been realized in maximizing sugar yields at very low cost. This review highlights literature on the impact of key substrate and enzyme features that influence performance, to better understand fundamental strategies to advance enzymatic hydrolysis of cellulosic biomass for biologica...

Enzymatic hydrolysis of cellulosic biomass

Production of a cellulolytic enzyme system in mixed-culture solid-state fermentation of soybean hulls supplemented with wheat bran

Production of Trichoderma cellulase in laboratory and pilot scale

Trichoderma reesei QM 9414 was grown on wheat straw as the sole carbon source. The straw was pretreated by physical and chemical methods. The particle size of straw was less than 0.177 mm. Growth of T. reesei QM 9414 was maximal with alkali-pretreated straw whereas cellulase production was optimal when physically pretreated straw was used as substrate. Cellulase yields expressed as IU enzyme activity/g cellulose present in the cultures were considerably higher when alkali pretreatment of wheat straw was omitted. Cellulase yields of 666 IU/g cellulose for filter paper activity (FPA) are the highest described for cultures of T. reesei QM 9414 carried out in analogous conditions. Crystallinity index of the cellulose contained in wheat straw increased slightly after alkali pretreatment. This increase did not decrease cellulose accessibility to the fungus. Delignification of wheat straw was not necessary to achieve the best cellulase production.

Enhanced cellulase production from Trichoderma reesei QM 9414 on physically treated wheat straw

Trichoderma reesei QM 9414 was grown in batch fermentation on wheat straw pretreated by different methods as the sole carbon source. Cellulase production was maximal with NaOH treated wheat straw at a concentration of 10 g/l and an initial pH of 5.5. The addition of fresh straw produced an elongation of the exponential phase or the beginning of a new exponential phase when the additions were carried out at 50 and 120 h, respectively. Filter paper and carboxymethylcellulase activities decreased as an answer to the addition of wheat straw and the levels were regained at the end of fermentation. The decreases of activities were accompanied by the increases of soluble sugar levels, which decreased at the end of fermentation. β-glucosidase activity was stimulated by wheat straw addition at 50 h while not by addition at 120 h; however, at the end of the fermentation the levels of activities were both similar to control. The studies of pH stabilities of these enzymes allow assurance that the effect of the addition of wheat straw on the enzyme activities is not produced by the changes of the pH during the fermentation.

Production of cellulases by Trichoderma reesei QM 9414 in batch and fed‐batch cultures on wheat straw

Adsorption of cellulase from Trichoderma reesei on wheat straw

Cellulase production by Trichoderma reesei on wheat straw

In this paper we studied the conditions for the production of β-glucosidase from T. reesei QM9414 in batch cultures using milled and sieved wheat straw as sole carbon source. High β-glucosidase production in the presence of wheat straw, a more realistic substrate than commercial cellulose, was obtained.



The influence of particle size of wheat straw on β-glucosidase production in cell-free, cell and cell-wall extracts was studied. The particle size of wheat straw notably influenced enzyme production in cell and extramycelial extracts but it was less important with respect to the cell wall bound enzyme.



β-glucosidase production was studied along of the fermentation. The results suggest a close relation between β-glucosidase from cell extract and extramycelial broth; geneticin levels of inhibition of β-glucosidase biosynthesis in both fractions were similar, a fact that suggests a common origin for the enzyme.



Kinetic parameters for β-glucosidase from cell free and cell extracts were Vmax = 0.28 μmol/min/mg, KM = 0.91 mM and Vmax = 0.095 μmol/min/mg, KM = 0.39 mM respectively. Kinetic parameters for β-glucosidase from cell-wall could not be calculated because experimental data did not fit the different monosubstrate equations.

D. Romero

Papers

Enhanced cellulase production from Trichoderma reesei QM 9414 on physically treated wheat straw

Production of cellulases by Trichoderma reesei QM 9414 in batch and fed‐batch cultures on wheat straw

Adsorption of cellulase from Trichoderma reesei on wheat straw

Cellulase production by Trichoderma reesei on wheat straw

β-Glucosidase production by Trichoderma reesei QM9414 on wheat straw. Location and some kinetic features