Enzymic saccharification of sugar beet pulp

Culture filtrates of Talaromyces emersonii UCG 208 grown on beet pulp can convert the polysaccharide components of this agricultural waste to soluble sugars. The saccharification process is facilitated if the pulp is milled or incubated with alkali or peracetic acid before addition of enzyme. However, treatment of unmilled pulp with commercial pectinase prior to incubation with Talaromyces filtrate is also very effective; under suitable conditions, complete hydrolysis of total polysaccharides has been achieved.     

Saccharification and protein enrichment of sugar beet pulp byPleurotus florida

Summary Pleurotus florida was grown in submerged culture with different concentrations of sugar beet pulp with a view to its nutritional upgrading. Micelial growth, protein enrichment and dry weight loss of the solid residues of cultures, as well as the evolution of reducing sugars in the liquid medium, were followed for 14 days. A product with 45% (w/w) protein and a saccharification extent as high as 35% (w/w) were obtained after 7 days with 1% (w/v) sugar beet pulp.     

A new pilot reactor for solid-state fermentation: Application to the protein enrichment of sugar beet pulp

A new pilot reactor for solid-state fermentation has been used for single-cell protein production on raw sugar beet pulp with a mutant, Trichoderma viride T.S. This pilot plant, having a maximum working capacity of one ton (ca. 200 kg dry matter) can be scaled up to the production plant level. During the process, the protein content increases from 9 to 20-21% (on the basis of dry matter) in 48 h. A material and heat balance is presented in relation with temperature and moisture level regulation during the process.     

Simultaneous saccharification and fermentation of pretreated sugar cane leaves to ethanol

The simultaneous saccharification and fermentation (SSF) of pretreated sugar cane leaves to produce ethanol using a cellulolytic enzyme complex from Trichoderma reesei QM 9414 and Saccharomyces cerevisiae NRRL-Y-132 was optimized. Enzymic saccharification parameters were evaluated prior to SSF studies. A 92% conversion of 2·5% substrate (alkaline hydrogen peroxide pretreated) to sugars was achieved at 50°C and pH 4·5, using T. reesei cellulase (40 FPU/g substrate), in 48 h. The pretreated substrate was then subjected to an SSF process using the cellulase complex and S. cerevisiae cells. Optimization of the SSF system is described.     

Selection of preculture conditions for solid state fermentation of sugar beet pulp

Summary For the protein upgrading of sugar-beet pulp in solid state fermentation byTrichoderma reesei andFusarium oxysporum, serveral conditions were studied to prepare an economical preculture for large scale process. The best performance was shown by a preculture obtained in 24 h from 1.5 % molasses solution at pH 4.5–5.0 with 1.0 % milled beet pulp.     

A pilot process of solid state fermentation from sugar beet pulp for the production of microbial protein

The bioconversion of sugar beet pulp (SBP) into microbial protein by the process of solid state fermentation (SSF) was studied. Two SSF reactors (each having overall dimensions of 17.6 × 3.6 × 2.0 m) with appropriate control systems were constructed. This pilot plant has a maximum working capacity of 50 metric tons (20% dry metter). The production strain used was Aspergillus tamarii 827, which was isolated by our laboratory. The protein content of the products amounted to 22.4% in 48 h.     

Enzymatic hydrolysis of sugar beet pulp

Summary In order to prepare fermentable sugars from cellulosic raw materials, we have tested the effect of different pretreatments (heat, chemicals, pectinase) on enzymatic hydrolysis of sugar beet pulp. Best results (90% hydrolysis) have been obtained after heat treatment (30 mn at 120°C) with 16 IU.Fp.g^–1 of fresh pulp in 48 hours.     

Anaerobic biodegradation of sugar beet pulp

Sugar beet pulp is a by-product of sugar production and consists mainly of cellulose, hemicellulose and pectin. Its composition is suitable for biological degradation. A possible alternative for the utilization of this material (besides cattle feeding) can be anaerobic methanogenic degradation. It has an additional advantage – biogas production. Beet pulp was treated by a two-step anaerobic process. The first step consisted of hydrolysis andacidification. The second step was methanogenesis. In this paper, observation ofthe process of anaerobic degradation and determination of optimal parameters is discussed. A laboratory-scale model for sugar beet pulp anaerobic biodegradation was operated. Results of model performance have shown very good pulp digestion characteristics. In addition, high efficiency removal of organic matter was achieved. Methane yield was over 0.360 m³ kg^-1 dried pulp and excess sludge production was 0.094 g per gram COD added.     

Simultaneous saccharification and fermentation of lime-treated biomass

Simultaneous saccharification and fermentation (SSF) was performed on lime-treated switchgrass and corn stover, and oxidatively lime-treated poplar wood to determine their compatibility with Saccharomyces cerevisiae. Cellulose-derived glucose was extensively utilized by the yeast during SSF. The ethanol yields from pretreated switchgrass, pretreated corn stover, and pretreated-and-washed poplar wood were 72%, 62% and 73% of theoretical, respectively, whereas those from -cellulose were 67 to 91% of theoretical. The lower ethanol yields from treated biomass resulted from lower cellulose digestibilities rather than inhibitors produced by the pretreatment. Oxidative lime pretreatment of poplar wood increased the ethanol yield by a factor of 5.6, from 13% (untreated) to 73% (pretreated-and-washed).     

Simultaneous saccharification and fermentation of kraft pulp by recombinant Escherichia coli for phenyllactic acid production

Simultaneous saccharification and fermentation (SSF) of renewable cellulose for the production of 3-phenyllactic acid (PhLA) by recombinant Escherichia coli was investigated. Kraft pulp recovered from biomass fractionation processes was used as a model cellulosic feedstock and was hydrolyzed using 10–50 filter paper unit (FPU) g⁻¹ kraft pulp of a commercial cellulase mixture, which increased the glucose yield from 21% to 72% in an enzyme dose-dependent manner. PhLA fermentation of the hydrolyzed kraft pulp by a recombinant E. coli strain expressing phenylpyruvate reductase from Wickerhamia fluorescens TK1 produced 1.9 mM PhLA. The PhLA yield obtained using separate hydrolysis and fermentation was enhanced from 5.8% to 42% by process integration into SSF of kraft pulp (20 g L⁻¹) in a complex medium (pH 7.0) at 37 °C. The PhLA yield was negatively correlated with the initial glucose concentration, with a five-fold higher PhLA yield observed in culture medium containing 10 g L⁻¹ glucose compared to 100 g L⁻¹. Taken together, these results suggest that the PhLA yield from cellulose in kraft pulp can be improved by SSF under glucose-limited conditions.     

Pectinolytic activity ofClostridium thermocellum: Its use for anaerobic fermentation of sugar beet pulp

Summary Clostridium thermocellum is well known for its ability to convert cellulose into ethanol and to hydrolyse hemicellulose. The present work shows its ability to hydrolyse model pectins and to use them for growth. The main products on these substrates as well as on sugar beet pulps were as follows: acetate, ethanol and methanol. Galacturonase and lyase activities were measured in the fermentation broths. As shown by the accumulation of methanol in the medium, there is a pectin esterase activity but this one seems to be very low.     

Addendum: Recovery of cellulases from spent broth of sugar beet pulp fermentation byTrichoderma reesei

Summary The exo- and endo-glucanases ofTrichoderma reesei were recovered after growth on sugar beet pulp by vacuum concentration followed by (NH₄)₂SO₄ or acetone precipitation. Final respective recoveries were 53% and 57% for the exoglucanase and 41% and 42% for the endoglucanase. The resuspended acetone precipitates of both enzymes showed no loss of activity after 30 d at 4°C, pH 4.8 and in the presence of 0.5% sorbic acid. (NH₄)₂SO₄-precipitated exoglucanase lost activity in the same period.

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Resumen Las exo y endoglucanasas deTrichoderma reesei se recuperaron después de su crecimiento en pulpa de remolacha azucarera mediante concentración al vacío seguida de precipitación con (NH₄)₂SO₄ o acetona. La recuperación final fue de 53% y 57% para la exoglucanasa y de 41% y 42% para la endoglucanasa. La resuspensión de los precipitados cetónicos de ambos enzimas no mostró perdidas de actividad después de 30 d a 4°C y pH 4.8 en presencia de ácido sórbico. La exoglucanasa precipitada con (NH₄)₂SO₄ perdió actividad durante este mismo periodo.

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Résumé Les exo- et endoglucanases deTrichoderma reesei ont été récupérées après croissance sur pulpe de betterave sucrière, par concentration sous vide après précipitation par le sulfate ammonique ou l'acétone. Les récupérations ont été respectivement de 53% et 57% pour l'exoglucanase et de 41% et 42% pour l'endoglucanase. Les resuspensions des deux enzymes après précipitation à l'acétone n'ont révélé aucune perte d'activité après 30 jours à 4°C, pH 4.8 et en présence de 0.5% d'acide sorbique. L'exoglucanase précipitée au sulfate ammonique avait perdu toute activité dans le même laps de temps.

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(Based on a paper presented at the First Latin American Congress on Biotechnology, Tucumán, Argentina, October 4–8, 1987)     

Simultaneous saccharification and extractive fermentation of cellulosic substrates

Alcohol fermentation has traditionally been carried out in aqueous environments because of the ready solubility of reactant (sugar) and product (ethanol). However, extraction of the product ethanol into a nonmiscible phase can result in kinetic benefits due to reduced inhibition of the fermentation reactions. In this study, we report the development of a novel simultaneous saccharification and extractive fermentation (SSEF) process. Ethanol productivity was increased by up to 65% over conventional (nonextractive) fed-batch simultaneous saccharification systems when calculated on the basis of aqueous phase volume. The amount of water required for SSEF reactions was dramatically reduced from that required for conventional SSF. In batch SSEF reactors with 2.5% aqueous phase, 50% conversion of 25% (aqueous phase concentration) Solka Floc could be achieved in 48 h using 2 FPU/g cellulase. (c) 1996 John Wiley & Sons, Inc.     

Decolorization of sugar syrups using commercial and sugar beet pulp based activated carbons

Sugar syrup decolorization was studied using two commercial and eight beet pulp based activated carbons. In an attempt to relate decolorizing performances to other characteristics, surface areas, pore volumes, bulk densities and ash contents of the carbons in the powdered form; pH and electrical conductivities of their suspensions and their color adsorption properties from iodine and molasses solution were determined. The color removal capabilities of all carbons were measured at 1/100 (w/w) dosage, and isotherms were determined on better samples. The two commercial activated carbons showed different decolorization efficiencies; which could be related to their physical and chemical properties. The decolorization efficiency of beet pulp carbon prepared at 750 degrees C and activated for 5h using CO2 was much better than the others and close to the better one of the commercial activated carbons used. It is evident that beet pulp is an inexpensive potential precursor for activated carbons for use in sugar refining.     

Simultaneous saccharification and fermentation of cellulose to lactic acid

Recent interest in the industrial manufacture of ethanol and other organic chemicals from biomass has led to the utilization of surplus grain and cane juice as a fermentation feedstock. Since those starting materials are also foods, they are expensive. As an alternative, cellulosic substances-the most abundant renewable resources on earth(1)-have long been considered for conversion to readily utilizable hydrolyzates.(2, 3)For the production of ethanol from cellulose, we have proposed the simultaneous saccharification and fermentation (SSF) process.(4) In SSF, enzymatic cellulose hydrolysis and glucose fermentation to ethanol by yeast proceed simultaneously within one vessel. The process advantages-reduced reactor volume and faster saccharification rates-have been confirmed by many researchers.(5-8) During SSF, the faster saccharification rates result because the glucose product is immediately removed, considerably diminishing its inhibitory effect on the cellulase system.(9)To effectively apply the SSF method to produce substances fermented from glucose, several conditions should be satisfied. One is coincident enzymatic hydrolysis and fermentation conditions, such as pH and temperature. The other is that cellulase inhibition by the final product is less than that by glucose and/or cellobiose. One of us has reported that acetic acid, citric acid, itaconic acid, alpha-ketoglutaric acid, lactic acid, and succinic acid scarcely inhibit cellulase.(10) This suggests that if the microorganisms which produce these organic acids were compatible with cellulase reaction conditions, the organic acids could be produced efficiently from cellulosic substrates by SSF.In this article, the successful application of SSF to lactic acid production from cellulose is reported. Though there have been several reports of direct cellulose conversion to organic acids by anaerobes such as Clostridium, only trace amounts of lactic acid were detected in the fermentation medium among the low-molecular-weight fatty acid components.(11-13) Lactic acid is one of the most important organic acids and has a wide range of food-related and industrial applications.     

Continuous anaerobic conversion of sugar beet pulp to biogas

Summary A continuous two step anaerobic digestion of sugar beet pulp as carbon and energy source was carried out. The loading rates in the acidification reactor were varied from 5 g·1^–1·d^–1 to 15 g·1^–1·d^–1 while the hydraulic retention time was in the range of 10 hours to 30 hours; the corresponding values for the methane reactor varied from 2 days to nearly 6 days.With this reactor configuration a biogas yield of more than 85% of the theoretical value for total carbon conversion was achieved resulting in a corresponding COD reduction.     

Oxidative cross-linking of pectic polysaccharides from sugar beet pulp

Oxidative cross-linking of three beet pectin extracts with hydrogen peroxide/peroxidase resulted in an increase in viscosity at low concentrations and in the formation of a gel at higher concentrations. Gels were formed using concentrations of 1.5% for an autoclave preparation and one obtained by an acid extraction and of 3% for a second autoclaved extract. It was shown that in the autoclave extracts only rhamnogalacturonans and possibly the arabinans participated in the cross-linking reaction. Cross-linking of the autoclave extracts with ammonium persulfate resulted in a decrease in reduced viscosity and molecular weight, although ferulic acid dehydrodimers were formed. Treatment of the acid extracted pectin with ammonium persulfate gave a slow increase in viscosity and the formation of a high-molecular-weight population was observed. For both oxidative systems, the 8-5 dehydrodimer was predominant after cross-linking.     

Fractionation of sugar beet pulp by introducing ion-exchange groups

Sugar beet pulp (SBP) was chemically modified with the goal to utilize this method for the preparation of water-soluble polysaccharides. Yields of the trimethylammoniumhydroxypropylated (TMAHP) polysaccharide fractions prepared under vacuum in absence of NaOH or KOH, as well as their molar masses, were higher than those obtained when the samples were only extracted with hydroxide at ambient pressure. The hydroxypropylsulfonated (HPS) fractions extracted at ambient pressure with NaOH had higher yields and molar mass than the TMAHP fractions extracted under vacuum with KOH. Introduction of both ion-exchange groups in one step under vacuum with NaOH gave better yields than with KOH, but smaller than for the solely sulfonated sample. The molar masses of chemically modified fractions were smaller than those extracted only with alkali or water. According to the monosaccharide composition all the fractions are mixtures of arabinogalactans and rhamnogalacturonoans.     

Effect of pig faecal donor and of pig diet composition on in vitro fermentation of sugar beet pulp

Two experiments were undertaken to investigate the influence of (1) pig bodyweight and (2) dietary fibre content of the diet on the in vitro gas production of sugar beet pulp fibre using faecal inoculum.In the first experiment, inocula prepared from young pigs (Y; 16–50 kg), growing pigs (G; 62–93 kg) and sows (S; 216–240 kg) were compared. Sugar beet pulp, hydrolysed in vitro with pepsin and then pancreatin, was used as the fermentation substrate. The cumulated gas productions over 144 h were modelled and the kinetics parameters compared. Lag times (Y: 4.6 h; G: 6.4 h; S: 9.2 h) and half-times to asymptote (Y: 14.7 h; G: 15.9 h; S: 20.8 h) increased with pig bodyweight (P<0.001) and the fractional degradation rates of the substrate differed between the pig categories (Y: 0.110 h⁻¹; G: 0.115 h⁻¹; S: 0.100 h⁻¹; P<0.001). The final gas production was not affected (P=0.10) by the inoculum source.In the second experiment hydrolysed sugar beet pulp was fermented with four inocula prepared from pigs fed diets differing in their total and soluble dietary fibre contents, i.e. low fibre diet rich in soluble fibre (LOW-S) or in insoluble fibre (LOW-I) or high fibre diet rich in soluble fibre (HIGH-S) or in insoluble fibre (HIGH-I). The total and the soluble dietary fibres influenced the kinetics of gas production. The presence of soluble fibres decreased the lag times, whatever the total dietary fibre content (2.7 h for LOW-S versus 3.5 h for LOW-I, 4.0 h for HIGH-S versus 4.4 h for HIGH-I; P<0.001). The half-times to asymptote were higher with the low fibre diets (P<0.001) and, for similar total dietary fibre contents, they were lower when the proportion of soluble fibres increased (LOW-S: 9.9 h; LOW-I: 11.4 h; HIGH-S: 8.9 h; HIGH-I: 10.1 h; P<0.001). The fractional degradation rates of the substrate were the highest with the fibre-rich diet containing a high proportion of soluble fibres (0.158 h⁻¹; P<0.001).In conclusion, the bodyweight of the faeces donors and the dietary fibre composition of the pig diet influence the in vitro fermentation kinetics of hydrolysed sugar beet pulp, but not the final gas production.