Organosolv pretreatment for enzymatic hydrolysis of poplars: I. Enzyme hydrolysis of cellulosic residues

Aspen (Populus tremuloides) and black cottonwood (Populus trichocarpa) organosolv pulps produced in a wide range of solvent composition (between 30 and 70% by volume of methanol) and catalysts (H(2)SO(4) and H(3)PO(4)) such that the cooking liquor pH 相似文献   

Differential speed two roll mill pretreatment of cellulosic materials for enzymatic hydrolysis

Differential speed two roll milling is an effective pretreatment for increasing the susceptibility of cellulose to enzymatic hydrolysis. Using mills with three, six, and ten in. diam rolls and processing times of 10 min or less results in the following percent increases in susceptibility over untreated controls: cotton, 1100; maple chips, 1600; white pine chips, 600; newspaper, 125. In comparison, ball milling of newspaper for 24 hr gives only a 62% increase. A further advantage of the roll mill is the increased wet density of the product permitting higher slurry concentrations during hydrolysis. Important parameters of mill effectiveness are roll clearance and processing time.     

Partial acid hydrolysis of cellulosic materials as a pretreatment for enzymatic hydrolysis

Partial acid hydrolysis was studied as a per treatment to enhance enzymatic hydrolysis, such a pretreatment was carried out in a continuous flow reactor on oak corn Stover, newsprint, and Solka Floc at temperatures ranging from 160 to 220°C, acid concentration ranging from 0 to 1.2%, and a fixed treatment time of 0.22 min. The resulting slurries and solids were than hydrolyzed with Trichoderma ressei QM 9414 cellulase at 50°C for 48 hr. For all substrates except Solka Floc, increased glucose yields were achieved during enzymatic hydrolysis of the pretreated materials as compared to hydrolysis of the original substrate. In several cases, after pretreatment, 100° of the potential glucose content of the substrate was converted to glucose after 24hr of enzymatic hydrolysis. It is felt that the increased glucose yields achieved after this pretreatment are due to acid's removal of hemicellulose, reduced degree of polymerization, and possibly due to a change in the crystal structure of the cellulose.     

Microbial cultures in the utilization of cellulosic materials

This review elaborates on the most recent microbial development in saccharification of cellulose and cellulase formation. A particular highlight is a new genetic-immunochemical approach investigating the mechanism of adhesion of bacterial cellulase to cellulose during cellulose conversion. New developments and recent reviews in hemicellulose and lignin degradation are also covered.     

Fibrillation tendency of cellulosic fibers—Part 4. Effects of alkali pretreatment of various cellulosic fibers

The effects of alkali type and the concentration in the alkali treatments on the weight loss in six cellulosic fibers and their influences on the fibrillation tendency were investigated. The fibril number of the cellulosic fibers pretreated with alkalis (FN_pre) increased with increasing the alkali concentrations as well as the weight loss of the fiber except in the lyocell fiber treated in NaOH and KOH solutions. The FN_pre in lyocell was reduced as the fibers were treated in 5 mol/l NaOH and KOH solutions. This result and the fact that the fibers were split in organic alkali such as tetramethylammonium hydroxide even at the low weight loss suggested that not only the loss of cellulose component but also reorganization of microfibril structure, inhomogeneous swelling of the fibers and other influences control the fibrillation tendency of cellulosic fibers.     

Evaluation of some newer non-steroidal anti-inflammatory indan-1-acids in various biological systems.

In line of the effort towards development of some newer indanyl non-steroidal anti-inflammatory agents and providing comprehensive SAR among this class of compounds some significantly active derivatives with low ulcerogenic potential were identified. Dealing with various long chain and branched chain compounds among this series, 3-(5, 6-dimethoxy indan-1-yl) propionic acid, 2-(5, 6-dimethoxy indan-1-yl) propionic acid and 3-(6-methoxy indan-1-yl) propionic acid were observed to have encouraging biological activity. Screening in various animal models of inflammation suggests their longer duration of action and lower ulcerogenic liability.     

Enhancement of methane production from cassava residues by biological pretreatment using a constructed microbial consortium

In the study, a stable thermophilic microbial consortium with high cellulose-degradation ability was successfully constructed. That several species of microbes coexisted in this consortium was proved by DGGE (denaturing gradient gel electrophoresis) and sequence analysis. The cooperation and symbiosis of these microbes in this consortium enhanced their cellulose-degradation ability. The pretreatment of cassava residues mixing with distillery wastewater prior to anaerobic digestion was investigated by using this microbial consortium as inoculums in batch bioreactors at 55 °C. The experimental results showed that the maximum methane yield (259.46 mL/g-VS) of cassava residues was obtained through 12 h of pretreatment by this microbial consortium, which was 96.63% higher than the control (131.95 mL/g-VS). In addition, it was also found that the maximum methane yield is obtained when the highest filter paper cellulase (FPase), carboxymethyl cellulase (CMCase) and xylanase activity and soluble COD (sCOD) are produced.     

Reactor scale up for biological conversion of cellulosic biomass to ethanol

The absence of a systematic scale-up approach for biological conversion of cellulosic biomass to commodity products is a significant bottleneck to realizing the potential benefits offered by such conversion. Motivated by this, we undertook to develop a scale-up approach for conversion of waste paper sludge to ethanol. Physical properties of the system were measured and correlations were developed for their dependence upon cellulose conversion. Just-suspension of solid particles was identified as the scale up criterion based on experiments at lab scale. The impeller speed for just solids suspension at large scale was predicted using computational fluid dynamics simulations. The scale-up strategy was validated by analyzing mixing requirements such as solid–liquid mass transfer under the predicted level of agitation at large scale. The scale-up approach enhances the prediction of reactor performance and helps provide guidelines for the analysis and design of large scale bioreactors based on bench scale experimentation.     

Improving utilization of crop residues. Evaluation of two chemicals used to improve digestibility of crop residues

Two chemical treatments involving chelating metal caustic swelling (CMCS) and sodium hydroxide (NaOH) were evaluated for their ability to affect in vitro dry matter and in vitro cellulose disappearance (IVDMD and IVCD, respectively), dry matter disappearance in sacco and the chemical composition of two low-quality crop residues, cornstalks (CS) and soya bean residue (SBR). At $\text{chemical}\text{substrate}$ ratios ranging from 0.25:1 to 5:1, linear increases in IVCD of both roughages were noted. Improvements in IVCD of CS and SBR were noted at 1:1, 3:1 and 5:1$\text{chemical}\text{substrate}$ ratios, regardless of whether the treated roughages were used immediately after treatment (fresh basis) or preserved by drying or freezing. Decreased concentrations of neutral detergent fibre (NDF), acid detergent fibre (ADF), acid detergent lignin (ADL) and crude protein (CP) resulted from chemical treatment. No effects of the water component of the solvent solutions were observed. Dry matter disappearance in sacco of CMCS- and NaOH-treated roughages was increased at all $\text{chemical}\text{substrate}$ ratios tested. Treatment of either roughage with CMCS resulted in greater digestibility in sacco than did treatment with NaOH. Treatment of CS with CMCS resulted in higher rates of digestion than did treatment with NaOH. No differences in rates of digestion of SBR were noted between chemicals. Both chemicals were more effective in improving digestibility of CS than of SBR.     

Energy requirements and process design considerations in compression-milling pretreatment of cellulosic wastes for enzymatic hydrolysis

The effectiveness of compression-milling pretreatment of lignocellulosics for enzymatic hydrolysis has been demonstrated for a wide variety of substrate sources. Reductions in the degree of crystallinity and the degree of polymerization of cellulose and partial destruction of the structural integrity of lignocellulosics brought about by compression milling significantly increase the susceptibility of cellulose to enzymatic hydrolysis. The enzymatic hydrolysis yield was found to be directly related to the specific energy input to the cellulosic substrate (kWh/1b substrate) by compression milling, and the energy input can be controlled by the milling time. The enzymatic hydrolysis yeilds from cellulosic materials pretreated by compression milling also vary significantly depending on the source and kind, the composition milling also vary significantly depending on the source and kind, the composition (contents of lignin and other components), and the structure. The power requirements for compression milling which renders equivalent hydrolysis yields also depend on the source and kind of lignocellulosics to be pretreated. For newspaper, the specific energy input required for 55% sugar yield is estimated as 0.3 kWh/lb substrate including 15% power loss. The additional sugar yield gained from the enzymatic hydrolysis of compression-milled newspaper (over and above the sugar yield of untreated substrate) is determined as 453 g sugar/kWh energy input.     

Improvement of the biodegradation of some cellulosic wastes by acid pretreatment.

Acid pretreatment of cellulosic wastes improved their susceptibility to Fusarium acuminatum enzymes. The effectiveness of acid pretreatment was demonstrated with an increase in both fungal growth and enzyme activities. A growth yield of 0.15 g/100 ml was achieved on medium containing 5% acid pretreated pods of bean for 60 minutes. Avicelase (C1), carboxymethylcellulase (Cx) and B-glucosidase (C2) reached their maximal biosynthesis on acid pretreated wheat bran, sugar-cane bagasse and sawdust-containing media, respectively. Xylanase and pectinase attained their highest accumulation on pretreated pods of bean media. A mixture of free sugars has been released by acid pretreatment. O.199 g dry mycelium was obtained when the fungus was grown on 100 ml of medium containing hydrolysate of 10% H2SO4 pretreated pods of bean for 30 min. No cellulase enzymes could be detected on hydrolysate medium at the time that low contents of both xylanase and pectinase were accumulated.     

Enhanced production of cellulases byCellulomonas strains grown on different cellulosic residues

Cellulomonas strains consumed commercial cellulose, cellulosic residues, xylan, cellobiose and carboxymethyl cellulose (CMC) as carbon sources in liquid culture, the growth being the most on cellobiose medium. All three components of the cellulase complex ofCellulomonas were produced when the organisms utilized all substrates as sole carbon and energy sources. The filter-paper cellulase (FPase) and endo-glucanase (CMCase) activities were higher in media containing α-cellulose and cellulosic residues than in media containing CMC, cellobiose, and xylan. Cell-free supernatants of all organisms exhibited greater CMC hydrolyzing activity than filter paper and β-glucoside hydrolyzing activities. All strains synthesized β-glucosidase maximally on cellobiose followed by commercial cellulose and cellulosic residues.C. biazotea produced the highest FPase and CMCase activity during growth on α-cellulose. It was followed byC. flavigena, C. cellasea, andC. fimi. Endo-glucanase and FPase from all organisms were secreted into the medium; 10–13 % became adsorbed on the surface of the insoluble substrates and could be successfully eluted using Tween 80. β-Glucosidase was located in cell extracts from all organisms.C. biazotea produced FPase and β-glucosidase activities several-fold greater than those produced by many other strains ofCellulomonas and some other cellulolytic bacteria and fungi. These studies were supported byPakistan Atomic Energy Commission. Some chemicals were purchased from funds allocated byUnited States Agency for International Development, Washington (DC, USA), under PSTC proposal 6.163.     

Combinations of mild physical or chemical pretreatment with biological pretreatment for enzymatic hydrolysis of rice hull

Two novel two-step pretreatments for enzymatic hydrolysis of rice hull (RH) were proposed to lower the severity requirement of fungal pretreatment time. They consisted of a mild physical or chemical step (ultrasonic and H(2)O(2)) and a subsequent biological treatment (Pleurotus ostreatus). The combined pretreatments led to significant increases of the lignin degradation than those of one step pretreatments. After enzymatic hydrolysis of the pretreated RH, the net yields of total soluble sugar (TS) and glucose (G) increased greatly. The combined pretreatment of H(2)O(2) (2%, 48 h) and P. ostreatus (18 d) was more effective than sole pretreatment of P. ostreatus for 60 d. It could remarkably shorten the residence time and reduce the losses of carbohydrates. Ligninase analyses and SEM observations indicated that the enhancing of the efficiency could possibly attribute to the structure disruption of the RH during the first pretreatment step. So, the combined pretreatment could be recommended to different lignocellulosic materials for enzyme based conversions.     

Automatic methods for predicting functionally important residues

Sequence analysis is often the first guide for the prediction of residues in a protein family that may have functional significance. A few methods have been proposed which use the division of protein families into subfamilies in the search for those positions that could have some functional significance for the whole family, but at the same time which exhibit the specificity of each subfamily ("Tree-determinant residues"). However, there are still many unsolved questions like the best division of a protein family into subfamilies, or the accurate detection of sequence variation patterns characteristic of different subfamilies. Here we present a systematic study in a significant number of protein families, testing the statistical meaning of the Tree-determinant residues predicted by three different methods that represent the range of available approaches. The first method takes as a starting point a phylogenetic representation of a protein family and, following the principle of Relative Entropy from Information Theory, automatically searches for the optimal division of the family into subfamilies. The second method looks for positions whose mutational behavior is reminiscent of the mutational behavior of the full-length proteins, by directly comparing the corresponding distance matrices. The third method is an automation of the analysis of distribution of sequences and amino acid positions in the corresponding multidimensional spaces using a vector-based principal component analysis. These three methods have been tested on two non-redundant lists of protein families: one composed by proteins that bind a variety of ligand groups, and the other composed by proteins with annotated functionally relevant sites. In most cases, the residues predicted by the three methods show a clear tendency to be close to bound ligands of biological relevance and to those amino acids described as participants in key aspects of protein function. These three automatic methods provide a wide range of possibilities for biologists to analyze their families of interest, in a similar way to the one presented here for the family of proteins related with ras-p21.     

Reducing agents improve enzymatic hydrolysis of cellulosic substrates in the presence of pretreatment liquid

Enzymatic hydrolysis of pretreated lignocellulosic substrates has emerged as an interesting option to produce sugars that can be converted to liquid biofuels and other commodities using microbial biocatalysts. Lignocellulosic substrates are pretreated to make them more accessible to cellulolytic enzymes, but the pretreatment liquid partially inhibits subsequent enzymatic hydrolysis. The presence of pretreatment liquid from Norway spruce resulted in a 63% decrease in the enzymatic saccharification of Avicel compared to when the reaction was performed in a buffered aqueous solution. The addition of 15 mM of a reducing agent (hydrogen sulfite, dithionite, or dithiothreitol) to reaction mixtures with the pretreatment liquid resulted in up to 54% improvement of the saccharification efficiency. When the reducing agents were added to reaction mixtures without pretreatment liquid, there was a 13-39% decrease in saccharification efficiency. In the presence of pretreatment liquid, the addition of 15 mM dithionite to Avicel, α-cellulose or filter cake of pretreated spruce wood resulted in improvements between 25 and 33%. Positive effects (6-17%) of reducing agents were also observed in experiments with carboxymethyl cellulose and 2-hydroxyethyl cellulose. The approach to add reducing agents appears useful for facilitating the utilization of enzymes to convert cellulosic substrates in industrial processes.     

Effect of Fenton's pretreatment on cotton cellulosic substrates to enhance its enzymatic hydrolysis response

Fenton’s reagent that generates reactive hydroxyl radical species was evaluated for its effectiveness as a pretreatment agent on cotton cellulosic substrates to increase its susceptibility to cellulase enzyme. Response surface methodology was used to optimize four different process variables viz., time of reaction; substrate size and concentrations of Fe²⁺ and H₂O₂. Overall, the cellulose substrates treated at 0.5 mM concentration of Fe²⁺, 2% concentration of H₂O₂ for a reaction period of 48 h gave the highest enzyme activity as determined using the response surface methodology. Cellulose substrates with high aspect ratio recorded better enzyme response than that with low aspect ratio which is supported by copper number estimation. The cellulosic substrate prepared using a combination of optimized Fenton’s pretreatment conditions and/or enzyme hydrolysis were studied and characterized by atomic force microscopy and scanning electron microscopy. Additionally, degree of polymerization analysis gives further insight into the degradation during Fenton’s reaction.     

Substrate utilization kinetic model for biological treatment process

The applicability of Contois' kinetic equation to aerobic and anaerobic treatments of organic wastes is investigated. A refractory coefficient to account for the nonbiodegradable portion of the organic substrates in the digester is incorporated into the kinetic equation. The kinetic equation is applied to the data for aerobic digestions of organic substrates and for anaerobic treatment of dairy wastes. They all show a very good fit of the kinetic equation to the data. Furthermore, the kinetic parameters and the refractory coefficients are shown to be independent of influent organic substrate concentration. This study confirms previous reports that the effluent quality of biological treatment systems for organic wastes depends on influent organic waste concentration. The effect of temperature on the kinetic parameters and the refractory coefficient for anaerobic treatment of sewage sludge are studied. It shows that the kinetic parameters vary with temperature, while the refractory coefficient remains fairly constant. Equations to predict biodegradable treatment efficiency and volumetric substrate utilization rate are also briefly discussed.     

Statistical optimization of sulfite pretreatment of corncob residues for high concentration ethanol production

In this study, a central composite design of response surface method was used to optimize sulfite pretreatment of corncob residues, in respect to sulfite charge (5-10%), treatment time (1-2h), liquid/solid (l/s) ratio (6:1-10:1) and temperature (150-180°C) for maximizing glucose production in enzymatic hydrolysis process. The relative optimum condition was obtained as follows: sulfite charge 7.1%, l/s ratio 7.6:1, temperature 156°C for 1.4h, corresponding to 79.3% total glucan converted to glucose+cellobiose. In the subsequent simultaneous saccharification and fermentation (SSF) experiments using 15% glucan substrates pretreated under this kind of conditions, 60.8 g ethanol l(-1) with 72.2% theoretical yield was obtained.