Cellulase enzyme production by various fungal strains on different carbon sources

In the process of making ethanol from lignocellulosics, enzyme production is still the most crucial and costly step. Trichoderma reesei Rut C-30 is the best known cellulase producer. This fungus is very good in endo- and exoglucanase production, however, the amount of β-glucosidase excreted is not sufficient for hydrolysis. In addition, T. reesei Rut C-30 is sensitive to inhibitors generated during steam-pretreatment of wood. In the present study, two good cellulase producers were selected from 16 fungal strains and investigated regarding filter paper activity (FPU) and β-glucosidase activity using inexpensive lignocellulosic carbon sources for cultivation. T. reesei Rut C-30 proved to be a good cellulase producer, resulting in a maximum FPU of 1.39 FPU ml Ś1 on washed, steam-pretreated willow, but the β-glucosidase activity was insufficient. High β-glucosidase activities were reached with T. viride OKI B1 on all substrates, with a maximum activity of 1.62 IU ml Ś1 on steam-pretreated spruce

Thermal behavior of native, washed and steam exploded lignocellulosic biomass samples

The aim of this study was to evaluate the chemical changes in the main components (cellulose, hemicellulose and lignin) of various lignocellulosic biomass samples during the steam explosion pretreatment. Pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) and thermogravimetry/mass spectrometry (TG/MS) measurements have been performed on different native, washed and steam exploded woody (willow and spruce) and herbaceous (hemp, wheat straw and sweet sorghum bagasse) biomass samples. The main differences between the thermal decomposition of the samples are interpreted in terms of the altered structure of the biomass samples by the effective steam explosion treatment and the different alkali ion contents which have been determined using inductively coupled plasma-optical emission spectroscopy (ICP-OES) method. In order to separate these two effects, the native biomass samples have been washed with hot water to remove the main parts of the potassium and sodium ions. The concentration of K+ and Na+ has been reduced in the treated biomass samples so the thermal decomposition mechanism has been altered due to the elimination of the catalytic effects. Principal Component Analysis (PCA) has been used to find statistical correlations between the data. The functional group compositions of the lignin molecules have been modified significantly as indicated by the pyrograms and the score plot of the PCA. The amount of hemicellulose has been reduced. On the other hand, the relative amount of the structurally modified cellulose has been increased in the samples by the steam explosion pretreatment step

Process considerations of a biorefinery producing value-added products from corn fibre

Corn fibre, a co-product of corn wet milling, can be a suitable raw material of a biorefinery producing biofuels and value-added chemicals. The simulated process is able to produce bioethanol, biomethane and xylitol synergistically, while it also covers its own heat demand. The proposed plant consists of the following process steps: fractionation, enzymatic hydrolysis and ethanol fermentation, distillation and dehydration, anaerobic digestion, biogas upgrading, aerobic waste water treatment, combined heat and power production, xylitol fermentation and recovery. Various scenarios of the biorefinery were investigated and the process configurations were compared in terms of energy efficiency, or mass flows of the products. Incineration of the sludge and production of district heat are found to be effective methods to increase the energy efficiency, on which aerobic sludge yield has a great effect. The solid-liquid separations, which are carried out in filterpress, have a curial role in terms of energy efficiency . Combustion of the solid part of cellulose hydrolysis residue is favourable compared with the anaerobic digestion, except if the dry matter content of the filterpressed solid was set to 30% instead of 40%. The amounts of the produced xylitol and biomethane are variable, which ensures the ability of market adaptation for the biorefinery

Evaluation of an online fermentation monitoring system

The need to introduce promising bioethanol production technologies calls for advanced laboratory techniques to study experiment designs and to obtain their results in a quick and reliable way. Real time monitoring based on general principles of ethanol fermentation, such as effluent CO2 volume, avoids time consuming steps, long lasting analyses and delivers information about the process directly. A device based on the above features and capable for real time monitoring on parallel channels was developed by the authors and is described in this paper. Both for calibration and for fermentation, test runs were carried out on different days and channels. Statistical evaluation was based on the obtained data. According to the t-test (P=0.05) and Grubbs analysis, the calibration method is reliable regardless of the date of calibration. When evaluating the fermentation results by ANCOVA acceptable standard derivations were obtained as impact of channel (58.8 ml), date (82.1 ml) and incorporating all impacts (116.2 ml). The final ethanol concentrations calculated based on the gas volume were compared to ones determined by HPLC and an average difference of 10% was found. Thus, the device proved to be advantageous in monitoring fermentation

Β-glucosidase production of two different Aspergillus strains

Β-glucosidase has an important role in cellulose degradation by cleaving the cellobiose to glucose units. Supplementation of Trichodermacellulase with exogenous β-glucosidase is needed to prevent the inhibition effect of cellobiose on exoglucanases and endoglucanases.Production of β-glucosidase by Aspergillus niger and Aspergillus phoenicishas been investigated under different fermentations. Both strains were appropriate for enzyme production at high level under the applied conditions. Cultivation of A. phoenicison Mandels' medium containing a complex nitrogen source has resulted in a higher β-glucosidase activity than on Vogel's medium. In an air-lift fermenter A. phoenicis grew in the shape of beads with 1.41 IU g–1 cell-associated enzyme activity. The fungal pellets can be used as in situ immobilized enzyme preparation. A. niger produced extracellular β-glucosidase at the level of 2.1 IU ml–1 in stirred-tank fermenter with a yield of 210 IU g–1 glucose and productivity of 21.8 IU l–1 h–1

Processing sweet sorghum into bioethanol - an integrated approach

Numerous evidences have been provided that juice of sweet sorghum and the leftover after squeezing, the bagasse can be a proper feedstock for bioethanol production. The possibility to integrate a side stream of sweet sorghum processing into the biomass-to-ethanol process was investigated in this study. The liquid fraction, a side stream of the necessary pretreatment of the bagasse was utilized as carbon source for Trichoderma reesei RUT-C30 to produce cellulase enzymes for biomass conversion. However, to overcome the inhibitory nature of the liquid fraction, pre-adaptation of the fungus on solid media was carried out previous to submerged fermentations. The results show that with this approach the lag phase caused by the inhibitors could be markedly shortened and an enhancement of the final enzyme production could be achieved when comparing the pre-adapted strains to reference

Ethanol production from mixtures of wheat straw and wheat meal

Background: Bioethanol can be produced from sugar-rich, starch-rich (first generation; 1G) or lignocellulosic (second generation; 2G) raw materials. Integration of 2G ethanol with 1G could facilitate the introduction of the 2G technology. The capital cost per ton of fuel produced would be diminished and better utilization of the biomass can be achieved. It would, furthermore, decrease the energy demand of 2G ethanol production and also provide both 1G and 2G plants with heat and electricity. In the current study, steam-pretreated wheat straw (SPWS) was mixed with presaccharified wheat meal (PWM) and converted to ethanol in simultaneous saccharification and fermentation (SSF). Results: Both the ethanol concentration and the ethanol yield increased with increasing amounts of PWM in mixtures with SPWS. The maximum ethanol yield (99% of the theoretical yield, based on the available C6 sugars) was obtained with a mixture of SPWS containing 2.5% water-insoluble solids (WIS) and PWM containing 2.5% WIS, resulting in an ethanol concentration of 56.5 g/L. This yield was higher than those obtained with SSF of either SPWS (68%) or PWM alone (91%). Conclusions: Mixing wheat straw with wheat meal would be beneficial for both 1G and 2G ethanol production. However, increasing the proportion of WIS as wheat straw and the possibility of consuming the xylose fraction with a pentose-fermenting yeast should be further investigated

Effects of pH and Aeration Conditions on Xylitol Production by Candida and Hansenula Yeasts

During the fermentative production of xylitol the following environmental parameters have a controlling effect on the xylitol yield: concentration of monosaccharides, temperature, aeration and pH. The purpose of the present work was to evaluate xylitol production by four yeast strains at different pH values and oxygen transfer rates (OTRs). The highest xylitol yields were obtained under the following conditions: Candida parapsilosis: pH 5.0, OTR 6.1 mmol L-1 h-1;Candida guilliermondii: pH 4.5, OTR 5.7 mmol L-1 h-1; Candida boidinii: pH 6.0, OTR 5.7 mmol L-1 h-1; Hansenula anomala: pH 4.5, OTR 2.8 mmol L-1 h-1 using 50 g L-1 initial xylose concentration

Integrated Process of Arabinose Biopurification and Xylitol Fermentation Based on the Diverse Action of Candida boidinii

Hemicellulosic hydrolysates of agro-residues are promising raw materials for xylitol and arabinose production through biotechnological methods. Two-step acidic fractionation of corn fibre was developed to produce a glucose- and arabinose-rich hydrolysate and a xylose-rich hydrolysate. An integrated process of arabinose biopurification on the glucose- and arabinose-rich hydrolysate and xylitol fermentation on the xylose-rich hydrolysate using Candida boidinii NCAIM Y.01308 was introduced, in which cell mass produced in arabinose biopurification was used as inoculum in the xylitol fermentation. Aerobic biopurification resulted in an arabinose solution containing 9.2 g L–1 of arabinose with a purity of 90 %, based on total sugars. Xylitol fermentation under microaerobic conditions resulted in a xylitol yield of 53 % of theoretical and a xylitol concentration of 10.4 g L–1 in three days. Hence, an integrated biorefinery process of hemicellulosic hydrolysates was developed based on the diverse action of C. boidinii to purify arabinose and produce xylitol