Thermo-mechanical extrusion pretreatment for conversion of soybean hulls to fermentable sugars

Thermo-mechanical extrusion pretreatment for lignocellulosic biomass was investigated using soybean hulls as the substrate. The enzyme cocktail used to hydrolyze pretreated soybean hulls to fermentable sugars was optimized using response surface methodology (RSM). Structural changes in substrate and sugar yields from thermo-mechanical processing were compared with two traditional pretreatment methods that utilized dilute acid (1% sulfuric acid) and alkali (1% sodium hydroxide). Extrusion processing parameters (barrel temperature, in-barrel moisture, screw speed) and processing aids (starch, ethylene glycol) were studied with respect to reducing sugar and glucose yields. The conditions resulting in the highest cellulose to glucose conversion (95%) were screw speed 350 rpm, maximum barrel temperature 80 °C and in-barrel moisture content 40% wb. Compared with untreated soybean hulls, glucose yield from enzymatic hydrolysis of soybean hulls increased by 69.6%, 128.7% and 132.2%, respectively, when pretreated with dilute acid, alkali and extrusion.     

Selected morphological and functional properties of extruded acetylated starch-cellulose foams

Starch acetates with degrees of substitution (DS) of 1.68 and 2.3 were extruded with 10%, 20% and 30% (w/w) cellulose and 20% (w/w) ethanol in a twin screw extruder at 150, 160 and 170 degrees C barrel temperatures and 170, 200 and 230 rpm screw speeds. X-ray diffractogram (XRD), differential scanning calormetry (DSC) and Fourier transform infrared spectroscopy (FTIR) were used to analyze the morphological properties of extruded foams. A central composite response surface design was applied to analyze the effects of starch type, cellulose content, barrel temperature and screw speed on specific mechanical energy requirement of extruding foams and the radial expansion ratio and compressibility of the extruded foams. XRD showed losses of DS starch and cellulose crystallinity and the formation of new complexes. FTIR spectra revealed that functional groups and chemical bonds were maintained after extrusion. Melting temperatures changed significantly when higher DS starch acetate was used. Cellulose content, barrel temperature and screw speed showed significant effects on thermal, physical and mechanical properties of extruded foams and the specific mechanical energy requirement.     

A Comparative Study on Torque Requirement During Extrusion Pretreatment of Different Feedstocks

Extrusion pretreatment of biomass can be one of the viable continuous pretreatment methods. The torque requirement of feedstock during extrusion was an important factor, and it was not reported in the literature. Screw compression ratio, screw speed, barrel temperature, and feedstock moisture content are the contributing factors to the torque. The current study was undertaken to investigate the effect of screw compression ratio, screw speed, temperature on torque requirement for different moisture content of switchgrass, prairie cord grass, corn stover, and big bluestem and to compare the torque requirement among the selected feedstocks. Biomass was extruded in a lab scale single-screw extruder with different screw compression ratios (2:1 and 3:1), screw speeds (50, 100, and 150?rpm), and barrel temperatures (50°C, 100°C, and 150°C) over a range of moisture contents (15%, 25%, 35%, and 45% wb). Statistical analyses revealed that all the independent variables considered in this study had a significant effect on torque requirement for the selected feedstocks. Among the independent variables considered moisture content, screw speed, and temperature had a negative effect on torque requirement for all the feedstocks. Switchgrass required the highest torque followed by corn stover, big bluestem, and prairie cord grass.     

自絮凝酵母SPSC01悬浮液的流变行为

用旋转黏度计测定了自絮凝颗粒酵母悬浮液的流变特性,并考察了其流变特性的影响因素,如菌体生物量、葡萄糖质量浓度、温度等。结果表明：自絮凝颗粒酵母悬浮液呈假塑性非牛顿流体,其流变特性服从幂律指数模型,随着菌体浓度的增大,稠度系数增大,流动行为指数减小;絮凝悬浮液的表观黏度随着糖浓度的增加有所增加,同一生物量下稠度系数k随着糖浓度的增加而增加,流动行为指数n随着糖浓度的增加变化很小,悬浮液的表观黏度随着温度的升高而降低;相同生物量下的流变指数随温度的升高而升高,而稠度系数随温度升高有所下降。     

SYNTHESIS OF CELLULOSE BY ACETOBACTER XYLINUM : VIII. On the Formation and Orientation of Bacterial Cellulose Fibrils in the Presence of Acidic Polysaccharides

The transfer of the glucosyl moiety from uridine diphosphate glucose in the presence of Acetobacter xylinum cell-free extracts led to the formation of a mixture of alkali-soluble and -insoluble cellodextrins. Typical cellulose fibrils could not be detected by electron microscopy in this product. Immediately after release into the medium, cellulose formed by whole cells is in a "prefibrous" form which passes through Millipore filters of 0.45 and 0.8 µ pore diameter. Non-filtrable fibrils arise from this material probably by a process of crystallization involving no extracellular enzymes. Fibrils formed in shaken cell suspensions intertwine and form aggregates visible to the naked eye. In quiet suspensions pellicles are formed which float on the surface. Soluble Na-carboxymethylcellulose (CMC) is incorporated into cellulose fibrils formed in its presence, probably by a process of co-crystallization. Aggregation of fibrils containing CMC is delayed because of electrostatic repulsion between carboxylic groups. The aggregation time depends on the amount of CMC incorporated, its degree of substitution, the pH of the medium, and the ionic strength. The amount of CMC incorporated depends on the relative concentration CMC/cellulose and on the similarity of the CMC and the cellulose molecules i.e. in molecular weight and the number of carboxyl substitutions. Cellulose pellicles formed in the presence of CMC by unshaken cell suspensions consist of crossed, superimposed layers of parallel oriented cellulose fibrils. The same phenomenon is observed when phosphomannan, but not levan, is substituted for CMC. The biogenesis of oriented cellulose fibrils is envisaged as a process comprising the following steps: polymerization of the monomeric precursor, diffusion of the molecule to crystallization sites, crystallization, and orientation. It is proposed that charged polysaccharides play a role similar to that of CMC in affecting the orientation of cellulose fibrils in the plant cell wall.     

Pretreatment of rice straw using an extrusion/extraction process at bench-scale for producing cellulosic ethanol

A combination of a twin-screw extrusion and an acid-catalyzed hot water extraction process performed at a bench-scale was used to prepare high monomeric xylose hydrolysate for cellulosic production. The influences of the screw speed (30-150 rpm), barrel temperature (80-160 °C) and corresponding specific mechanical energy of the extruder on the structural properties of the pretreated rice straw, sugar concentration and conversion were investigated. The optimal condition for the extrusion step was determined to be 40 rpm with 3% H2SO4 at 120 °C; the optimal condition for the extraction step was determined to be 130 °C for 20 min. After the pretreatment at the optimal condition, 83.7% of the xylan was converted to monomeric xylose, and the concentration reached levels of 53.7 g/L. Finally, after the subsequent enzymatic hydrolysis, an 80% yield of the total saccharification was obtained.     

Rheological,Mass Transfer,and Mixing Characterization of Cellulase-Producing Trichoderma reesei Suspensions

Steady and dynamic shear measurements are utilized to characterize the rheological behavior of Trichoderma reesei RUT-C30 fungal suspensions during batch growth on xylose (soluble substrate) or cellulose (particulate solid substrate) at three different fermentor impeller speeds (250, 400, and 550 rpm). Biomass concentrations versus time were unimodal on xylose and bimodal on cellulose. This behavior is consistent with relatively rapid, early growth on easily metabolized growth medium components (yeast extract), followed by a second, slower growth phase due to hydrolysis of recalcitrant cellulose by increasing cellulase concentrations. Critical dissolved oxygen (DO) concentration for T. reesei growth on cellulose was found to be 0.073 mmol/L. The DO was kept above this level by supplementing the air feed with pure oxygen, implying that mass transfer limitations were not the cause of bimodal cell growth. Steady shear rheological data showed shear thinning behavior and a yield stress for all broth samples regardless of substrate. Casson and Herschel−Bulkley constitutive equations fit steady shear data well. Dynamic shear measurements on broth suspensions indicated “gel-like” behavior at low strains, with microstructural breakdown at larger displacements. Time variations of the Casson model parameters (yield stress and Casson viscosity) and dynamic moduli (elastic and viscous modulus) followed both cell mass and morphology: a single maximum in all rheological variables resulted when cells were grown on xylose or on cellulose at impeller speeds of 400 or 550 rpm, and dual maxima were observed for cellulose-grown cells at 250 rpm.     

Macromolecular modifications of manioc starch components by extrusion-cooking with and without lipids

Macromolecular structure of manioc starch, extruded without and with lipids (oleic acid, dimodan, soya lecithin and copra) was studied, using chemical, enzymic, viscometric and chromatographic methods. Twin screw extrusion-cooking led to a macromolecular degradation of both amylose and amylopectin. The formation of lower molecular weight material was observed by a decrease of intrinsic viscosities of both components and also by their behaviour on Sepharose CL-2B, whereas no modification of β-amylolysis and iodine-binding capacity could be detected. The macromolecular degradation was increased by higher temperature and screw speed of the extruder, and was decreased by adding lipids during extrusion. Lipids such as fatty acids, mono- and triglycerides have been shown to act as lubricants (each type in its distinctive way). Lipid extraction, by different solvents, appears to have a low efficiency. Although the addition of triglycerides during extrusion reduces the macromolecular degradation, leading to a high solubility, the amylose-lipid complexes reduce the water-soluble fraction. This fraction was shown to be mainly composed of aggregated amylopectin-like material and to be highly stable after successive freeze-thaw cycles.     

Cellulase digestibility of pretreated biomass is limited by cellulose accessibility

Attempts to correlate the physical and chemical properties of biomass to its susceptibility to enzyme digestion are often inconclusive or contradictory depending on variables such as the type of substrate, the pretreatment conditions and measurement techniques. In this study, we present a direct method for measuring the key factors governing cellulose digestibility in a biomass sample by directly probing cellulase binding and activity using a purified cellobiohydrolase (Cel7A) from Trichoderma reesei. Fluorescence-labeled T. reesei Cel7A was used to assay pretreated corn stover samples and pure cellulosic substrates to identify barriers to accessibility by this important component of cellulase preparations. The results showed cellulose conversion improved when T. reesei Cel7A bound in higher concentrations, indicating that the enzyme had greater access to the substrate. Factors such as the pretreatment severity, drying after pretreatment, and cellulose crystallinity were found to directly impact enzyme accessibility. This study provides direct evidence to support the notion that the best pretreatment schemes for rendering biomass more digestible to cellobiohydrolase enzymes are those that improve access to the cellulose in biomass cell walls, as well as those able to reduce the crystallinity of cell wall cellulose.     

Pretreatments for Enhanced Enzymatic Hydrolysis of Pinewood: a Review

Pinewood is an abundant source of lignocellulosic biomass that has potential to be used as renewable feedstock in biorefineries for conversion into advanced biofuels and other value-added chemicals. However, its structural recalcitrance, due to the compact packing of its major components, viz. cellulose, hemicellulose and lignin, high lignin content, and high cellulose crystallinity, is a major bottleneck in its widespread use as a biorefinery feedstock. Typical chemical, thermal, and biological pretreatment technologies are aimed at removing lignin and hemicellulose fractions for improving enzyme accessibility and digestibility of cellulose. This review highlights common pine pretreatment procedures, associated key parameters and resulting enzymatic hydrolysis yields. The challenges and limitations are also discussed as well as potential strategies to overcome them, providing an essential source of information to realize pine as a compelling biorefinery biomass source.     

Features of promising technologies for pretreatment of lignocellulosic biomass

Cellulosic plant material represents an as-of-yet untapped source of fermentable sugars for significant industrial use. Many physio-chemical structural and compositional factors hinder the enzymatic digestibility of cellulose present in lignocellulosic biomass. The goal of any pretreatment technology is to alter or remove structural and compositional impediments to hydrolysis in order to improve the rate of enzyme hydrolysis and increase yields of fermentable sugars from cellulose or hemicellulose. These methods cause physical and/or chemical changes in the plant biomass in order to achieve this result. Experimental investigation of physical changes and chemical reactions that occur during pretreatment is required for the development of effective and mechanistic models that can be used for the rational design of pretreatment processes. Furthermore, pretreatment processing conditions must be tailored to the specific chemical and structural composition of the various, and variable, sources of lignocellulosic biomass. This paper reviews process parameters and their fundamental modes of action for promising pretreatment methods.     

Cellulose degradation and monitoring of viscosity decrease in cultures of Cellulomonas uda grown on printed newspaper

The rheological behavior of cultures of Cellulomonas uda with shredded printed newspaper as the carbon source was studied. The initial substrate concentrations ranged from 23 to 60 g/L. The changes in apparent viscosity were followed on-line by applying a commercially available process viscometer and discretely using a rotational viscometer with an anchor impeller. During the time of highest cellulose degradation, the broths exhibited a pseudoplastic behavior which could be explained satisfactorily by the power-law model. At the end of cultivation when cellulose degradation slowed down, the broths became Newtonian in behavior. Endo-1,4-beta-glucanase, 1,4-beta-xylanase, beta-glucosidase, and beta-xylosidase activities were also determined during cultivation as well as cellulose degradation and cell mass production. The beginning of endoglucanase formation and the start of the final viscosity decrease of the bacterial paper pulp suspensions could be correlated.     

Pretreatments to enhance the digestibility of lignocellulosic biomass

Lignocellulosic biomass represents a rather unused source for biogas and ethanol production. Many factors, like lignin content, crystallinity of cellulose, and particle size, limit the digestibility of the hemicellulose and cellulose present in the lignocellulosic biomass. Pretreatments have as a goal to improve the digestibility of the lignocellulosic biomass. Each pretreatment has its own effect(s) on the cellulose, hemicellulose and lignin; the three main components of lignocellulosic biomass. This paper reviews the different effect(s) of several pretreatments on the three main parts of the lignocellulosic biomass to improve its digestibility. Steam pretreatment, lime pretreatment, liquid hot water pretreatments and ammonia based pretreatments are concluded to be pretreatments with high potentials. The main effects are dissolving hemicellulose and alteration of lignin structure, providing an improved accessibility of the cellulose for hydrolytic enzymes.     

Rheology of corn stover slurries at high solids concentrations--effects of saccharification and particle size

The rheological characteristics of untreated and dilute acid pretreated corn stover (CS) slurries at high solids concentrations were studied under continuous shear using plate-plate type measurements. Slurry rheological behavior was examined as a function of insoluble solids concentration (10-40%), extent of pretreatment (0-75% removal of xylan) and particle size (-20 and -80 mesh). Results show that CS slurries exhibit shear-thinning behavior describable using a Casson model. Further, results demonstrate that the apparent viscosity and yield stress increase with increasing solids concentration (which corresponds to a decrease in free water). Dilute acid pretreatment leads to lower viscosity and yield stresses at equivalent solids concentrations, as does smaller particle size. Taken together, these findings are consistent with the hypothesis that the availability of free water in the slurry plays a significant role in determining its rheological behavior. In particular, as the free water content of the slurry decreases, e.g., with increasing solids concentration, the greater interaction among particles likely increases the apparent viscosity and yield stress properties of the slurry. The results also suggest that the availability of free water, and thereby slurry rheological properties, depend on the chemical composition of the corn stover as well as its physical characteristics such as particle size and porosity. Hydrophilic polymers within the cell wall, such as xylan or pectin, or larger pores within bigger particles, facilitate sequestration of water in the solid phase resulting in decreased availability of free water. Thus, dilute acid pretreated slurries, which contain smaller size particles having significantly lower xylan content than slurries of untreated milled stover, exhibit much lower viscosities and yield stresses than untreated slurries containing large particles at similar solid concentrations.     

Development of a helical-ribbon impeller bioreactor for high-density plant cell suspension culture

A double helical-ribbon impeller (HRI) bioreactor with a 11-L working volume was developed to grow high-density Catharanthus roseus cell suspensions. The rheological behavior of this suspension was found to be shear-thinning for concentrations higher than 12 to 15 g DW . L(-1). A granulated agar suspension of similar rheological properties was used as a model fluid for these suspensions. Mixing studies revealed that surface baffling and bottom profiling of the bioreactor and impeller speeds of 60 to 150 rpm ensured uniform mixing of suspensions. The HRI power requirement was found to increase singnificantly for agar suspensions higher than 13 g DW . L(-1), in conjunction with the effective viscosity increase. Oxygen transfer studies showed high apparent surface oxygen transfer coefficients (k(L)a approximately 4 to 45 h(-1)) from agar suspensions of 30 g DW . L(-1) to water and for mixing speeds ranging from 120 to 150 rpm. These high surface k(I)a values were ascribed to the flow pattern of this bioreactor configuration combined with surface bubble generation and entrainment in the liquid phase caused by the presence of the surface baffles. High-density C. roseus cell suspension cultures were successfully grown in this bioreactor without gas sparging. Up to 70% oxygen enrichment of the head space was required to ensure sufficient oxygen supply to the cultures so that dissolved oxygen concentration would remain above the critical level (>/=10% air saturation). The best mixing speed was 120 rpm. These cultures grew at the same rate ( approximately 0.4 d(-1)) and attained the same high biomass concentrations ( approximately 25 to 27 g DW . L(-1), 450 to 500 g filtered wet biomass . L(-1), and 92% to 100% settled wet biomass volume) as shake flask cultures. The scale-up potential of this bioreactor configuration is discussed.     

Rheological properties and gelation of aqueous cellulose-NaOH solutions

The shear rheology of a microcrystalline cellulose dissolved in a 9% NaOH aqueous solution was studied in the steady and oscillatory modes. The cellulose-(9% NaOH-H(2)O) mixtures show not to be true solutions. In the dilute regime, with cellulose concentration below 1%, the rheological behavior is typical of the one of suspensions. The formation of cellulose aggregates is favored when temperature is increased. In the semidilute regime, an irreversible aggregate-based gelation occurs, being faster with increasing temperature.     

Immobilization of cells by electrostatic droplet generation: a model system for potential application in medicine

The process of electrostatic extrusion as a method for cell immobilization was investigated that could be used for potential applications in medicine. An attempt was made to assess the effects of cell addition and polymer concentration on the overall entrapment procedure, ie, on each stage of immobilization: polymer-cell suspension rheological characteristics, electrostatic extrusion process, and the process ofgelation. The findings should contribute to a better understanding of polymer-cell interactions, which could be crucial in possible medical treatments. Alginate-yeast was used as a model system for carrier-cells. The electrostatic extrusion was considered as a complex two-phase flow system and the effects of cell and alginate concentrations on the resulting microbead size and uniformity were assessed. Under investigated conditions, microbeads 50-600 microm in diameter were produced and the increase in both alginate and cell concentrations resulted in larger microbeads with higher standard deviations in size. We attempted to rationalize the findings by rheological characterization of the cell-alginate suspensions. Rheological characterization revealed non-Newtonian, pseudoplastic behavior of cell-alginate suspensions with higher viscosities at higher alginate concentrations. However, the presence of cells even at high concentrations (5x10(8) and 1x10(9) cells/mL) did not significantly affect the rheological properties of Na-alginate solution. Lastly, we investigated the kinetics of alginate gelation with respect to the quantity of Ca2+ ions and cell presence. The gelation kinetics were examined under conditions of limited supply with Ca2+ ions, which can be essential for immobilization of highly sensitive mammalian cells that require minimal exposure to CaCl2 solution. The molar ratio of G units to Ca2+ ions of 3.8:1 provided complete crosslinking, while the increase in alginate concentration resulted in prolonged gelation times but higher strength of the resulting gel. The cell presence decreased the rate of network formation as well as the strength of the obtained Ca-alginate hydrogel.     

Two stage extrusion of plasticized pectin/poly(vinyl alcohol) blends

Blends of pectin with starch (high amylose and normal), poly(vinyl alcohol) (PVOH), and glycerol were extruded in a twin screw extruder, pelletized, and then further processed into blown film and extruded sheet using a single screw extruder. The samples were analyzed using tensile measurements, dynamic mechanical analysis, and scanning electron microscopy. PVOH levels of 24% or greater were necessary to successfully make blown film, while extruded sheet could be made at a level of 16% PVOH. Tensile strength and initial modulus of the extruded sheets were slightly higher in the machine direction than in the cross direction, while the reverse was true for elongation to break. For the blown films tensile strength tended to be higher in the transverse direction than in the machine direction, while the reverse was seen for initial modulus. Increased levels of PVOH led to increases in tensile strength and elongation to break, while initial modulus was decreased. Morphology as determined by SEM visually indicated stretching in the transverse direction of the blown films. The second stage extrusion appeared to promote -helix formation in the high amylose starch.     

Recovery of Sugars from Ionic Liquid Biomass Liquor by Solvent Extraction

The dissolution of biomass into ionic liquids (ILs) has been shown to be a promising alternative biomass pretreatment technology, facilitating faster breakdown of cellulose through the disruption of lignin and the decrystallization of cellulose. Both biological and chemical catalysis have been employed to enhance the conversion of IL-treated biomass polysaccharides into monomeric sugars. However, biomass-dissolving ILs, sugar monomers, and smaller carbohydrate oligomers are all soluble in water. This reduces the overall sugar content in the recovered solid biomass and complicates the recovery and recycle of the IL. Near-complete recovery of the IL and the holocellulose is essential for an IL-based pretreatment technology to be economically feasible. To address this, a solvent extraction technique, based on the chemical affinity of boronates such as phenylboronic acid and naphthalene-2-boronic acid for sugars, was applied to the extraction of glucose, xylose, and cellobiose from aqueous mixtures of 1-ethyl-3-methylimidazolium acetate. It was shown that boronate complexes could extract up to 90% of mono- and disaccharides from aqueous IL solutions, 100% IL systems, and hydrolysates of corn stover containing IL. The use of boronate complexes shows significant potential as a way to recover sugars at several stages in ionic liquid biomass pretreatment processes, delivering a concentrated solution of fermentable sugars, minimizing toxic byproducts, and facilitating ionic liquid cleanup and recycle.     

Rheology and Hydrodynamic Properties of Tolypocladium inflatum Fermentation Broth and Its Simulation

A physico-chemical, two phase simulated pseudoplastic fermentation (SPF) broth was investigated in which Solka Floc cellulose fibre was used to simulate the filamentous biomass, and a mixture of 0.1% (w/v) carboxymethyl cellulose (CMC) and 0.15 M aqueous sodium chloride was used to simulate the liquid fraction of the fermentation broth. An investigation of the rheological behaviour and hydrodynamic properties of the SPF broth was carried out, and compared to both a fungal Tolypocladium inflatum fermentation broth and a CMC solution in a 50 L stirred tank bioreactor equipped with conventional Rushton turbines. The experimental data confirmed the ability of the two phase SPF broth to mimic both the T. inflatum broth bulk rheology as well as the mixing and mass transfer behaviour. In contrast, using a homogeneous CMC solution with a similar bulk rheology to simulate the fermentation resulted in a significant underestimation of the mass transfer and mixing times. The presence of the solid phase and its microstructure in the SPF broth appear to play a significant role in gas holdup and bubble size, thus leading to the different behaviours. The SPF broth seems to be a more accurate simulation fluid that can be used to predict the bioreactor mixing and mass transfer performance in filamentous fermentations, in comparison with CMC solutions used in some previous studies.