Production of surfactin by Bacillus pumilus UFPEDA 448 in solid-state fermentation using a medium based on okara with sugarcane bagasse as a bulking agent

We studied the production of surfactin by Bacillus pumilus UFPEDA 448 in solid-state fermentation (SSF), using a medium based on okara with the addition of sugarcane bagasse as a bulking agent. The optimum proportions of okara and sugarcane bagasse were 50% each, by mass. Due to the relatively high production of proteases during SSF, pre-hydrolysis of the okara with a protease did not improve surfactin levels. The optimum temperature for surfactin production was 37 °C, with the incubation temperature affecting the ratios of the various surfactin homologues produced. Cultivation in column bioreactors with forced aeration under optimized conditions gave a surfactin level of 809 mg L^?1 of impregnating solution, which corresponds to 3.3 g kg-dry-solids^?1. This is the highest surfactin level that has been produced to date in SSF with a non-recombinant microorganism. The peak O₂ uptake rate was 20 mmol min^?1 kg-initial-dry-solids^?1, corresponding to metabolic waste heat production rate of 182 W kg-initial-dry-solids^?1. The tensioactive properties of the surfactin were similar to those reported in the literature for surfactin produced by submerged fermentation. These results suggest that it might be feasible to use SSF to produce surfactin but at large scale special attention will need to be given to heat removal.     

Fungal rock phosphate solubilization using sugarcane bagasse

The effects of different doses of rock phosphate (RP), sucrose, and (NH₄)₂SO₄ on the solubilization of RP from Araxá and Catal?o (Brazil) by Aspergillus niger, Penicillium canescens, Eupenicillium ludwigii, and Penicillium islandicum were evaluated in a solid-state fermentation (SSF) system with sugarcane bagasse. The factors evaluated were combined following a 2³?+?1 factorial design to determine their optimum concentrations. The fitted response surfaces showed that higher doses of RP promoted higher phosphorus (P) solubilization. The addition of sucrose did not have effects on P solubilization in most treatments due to the presence of soluble sugars in the bagasse. Except for A. niger, all the fungi required high (NH₄)₂SO₄ doses to achieve the highest level of P solubilization. Inversely, addition of (NH₄)₂SO₄ was inhibitory to P solubilization by A. niger. Among the fungi tested, A. niger stood out, showing the highest solubilization capacity and for not requiring sucrose or (NH₄)₂SO₄ supplementation. An additional experiment with A. niger showed that the content of soluble P can be increased by adding higher RP doses in the medium. However, P yield decreases with increasing RP doses. In this experiment, the maximal P yield (approximately 60?%) was achieved with the lower RP dose (3?g?L^?1). Our results show that SSF can be used to obtain a low cost biofertilizer rich in P combining RP, sugarcane bagasse, and A. niger. Moreover, sugarcane bagasse is a suitable substrate for SSF aiming at RP solubilization, since this residue can supply the C and N necessary for the metabolism of A. niger within a range that favors RP solubilization.     

Optimization of amylase production by Aspergillus niger in solid-state fermentation using sugarcane bagasse as solid support material

Synthesis of amylase by Aspergillus niger strain UO-01 under solid-state fermentation with sugarcane bagasse was optimized by using response surface methodology and empirical modelling. The process parameters tested were particle size of sugarcane bagasse, incubation temperature and pH, moisture level of solid support material and the concentrations of inoculum, total sugars, nitrogen and phosphorous. The optimum conditions for high amylase production (457.82 EU/g of dry support) were particle size of bagasse in the range of 6–8 mm, incubation temperature and pH: 30.2°C and 6.0, moisture content of bagasse: 75.3%, inoculum concentration: 1 × 10⁷ spores/g of dry support and concentrations of starch, yeast extract and KH₂PO₄: 70.5, 11.59 and 9.83 mg/g of dry support, respectively. After optimization, enzyme production was assayed at the optimized conditions. The results obtained corroborate the effectiveness and reliability of the empirical models obtained.     

Gas hold-up and oxygen mass transfer in three pneumatic bioreactors operating with sugarcane bagasse suspensions

Sugarcane bagasse is a low-cost and abundant by-product generated by the bioethanol industry, and is a potential substrate for cellulolytic enzyme production. The aim of this work was to evaluate the effects of air flow rate (Q _AIR), solids loading (%_S), sugarcane bagasse type, and particle size on the gas hold-up (ε _G) and volumetric oxygen transfer coefficient (k _L a) in three different pneumatic bioreactors, using response surface methodology. Concentric tube airlift (CTA), split-cylinder airlift (SCA), and bubble column (BC) bioreactor types were tested. Q _AIR and  %_S affected oxygen mass transfer positively and negatively, respectively, while sugarcane bagasse type and particle size (within the range studied) did not influence k _L a. Using large particles of untreated sugarcane bagasse, the loop-type bioreactors (CTA and SCA) exhibited higher mass transfer, compared to the BC reactor. At higher  %_S, SCA presented a higher k _L a value (0.0448 s^?1) than CTA, and the best operational conditions in terms of oxygen mass transfer were achieved for  %_S < 10.0 g L^?1 and Q _AIR > 27.0 L min^?1. These results demonstrated that pneumatic bioreactors can provide elevated oxygen transfer in the presence of vegetal biomass, making them an excellent option for use in three-phase systems for cellulolytic enzyme production by filamentous fungi.     

Water and water activity in the solid state fermentation of cassava starch by Aspergillus niger

Summary During the solid state fermentation (SSF) of cassava starch by Aspergillus niger estimations were made of total water, consumed water and the residual water remaining in small quantities after 23 h. A theoretical calculation based on the Ross equation showed that the water activity (a _w) of the substrate decreased to 0.85 towards the end of the culture. Such low values were assumed to be inhibitory to growth. The a _w of the substrate was increased when sugarcane bagasse was used as a high water retention capacity support. Higher growth rates and substrate conversion to biomass were obtained with this system, confirming that water availability is a critical factor in the SSF of starch substrates.Abbreviations A, B Experimental constants - a _w Water activity - H₂O_c Consumed water - H₂O_R Residual water - H₂O_T Total water - IDW Initial dry weight - IMC Initial moisture content - OUR Oxygen uptake rate - S Substrate dry weight - Sc Substrate conversion: consumed substrate/initial substrate - S _H Amount of sugars hydrolysed - SSF Solid state fermentation - X Biomass dry weight - W ^* Amount of solids/g of water     

Optimization of Aspergillus niger rock phosphate solubilization in solid-state fermentation and use of the resulting product as a P fertilizer

A biotechnological strategy for the production of an alternative P fertilizer is described in this work. The fertilizer was produced through rock phosphate (RP) solubilization by Aspergillus niger in a solid-state fermentation (SSF) with sugarcane bagasse as substrate. SSF conditions were optimized by the surface response methodology after an initial screening of factors with significant effect on RP solubilization. The optimized levels of the factors were 865 mg of biochar, 250 mg of RP, 270 mg of sucrose and 6.2 ml of water per gram of bagasse. At this optimal setting, 8.6 mg of water-soluble P per gram of bagasse was achieved, representing an increase of 2.4 times over the non-optimized condition. The optimized SSF product was partially incinerated at 350°C (SB-350) and 500°C (SB-500) to reduce its volume and, consequently, increase P concentration. The post-processed formulations of the SSF product were evaluated in a soil–plant experiment. The formulations SB-350 and SB-500 increased the growth and P uptake of common bean plants (Phaseolus vulgaris L.) when compared with the non-treated RP. Furthermore, these two formulations had a yield relative to triple superphosphate of 60% (on a dry mass basis). Besides increasing P concentration, incineration improved the SSF product performance probably by decreasing microbial immobilization of nutrients during the decomposition of the remaining SSF substrate. The process proposed is a promising alternative for the management of P fertilization since it enables the utilization of low-solubility RPs and relies on the use of inexpensive materials.     

Enhanced ethanol production from sugarcane juice by galactose adaptation of a newly isolated thermotolerant strain of Pichia kudriavzevii

The thermotolerant yeast strain isolated from sugarcane juice through enrichment technique was identified as a strain of Pichiakudriavzevii (Issatchenkiaorientalis) through molecular characterization. The P. kudriavzevii cells adapted to galactose medium produced about 30% more ethanol from sugarcane juice than the non-adapted cells. The recycled cells could be used for four successive cycles without a significant drop in ethanol production. Fermentation in a laboratory fermenter with galactose adapted P. kudriavzevii cells at 40 °C resulted in an ethanol concentration and productivity of 71.9 g L⁻¹ and 4.0 g L⁻¹ h⁻¹, respectively from sugarcane juice composed of about 14% (w/v) sucrose, 2% (w/v) glucose and 1% (w/v) fructose. In addition to ethanol, 3.30 g L⁻¹ arabitol and 4.19 g L⁻¹ glycerol were also produced, whereas sorbitol and xylitol were not formed during fermentation. Use of galactose adapted P. kudriavzevii cells for ethanol production from sugarcane juice holds potential for scale-up studies.     

Laccase induction by synthetic dyes in Pycnoporus sanguineus and their possible use for sugar cane bagasse delignification

The use of synthetic dyes for laccase induction in vivo has been scarcely explored. We characterized the effect of adding different synthetic dyes to liquid cultures of Pycnoporus sanguineus on laccase production. We found that carminic acid (CA) can induce 722 % and alizarin yellow 317 % more laccase than control does, and they promoted better fungal biomass development in liquid cultures. Aniline blue and crystal violet did not show such positive effect. CA and alizarin yellow were degraded up to 95 % during P. sanguineus culturing (12 days). With this basis, CA was selected as the best inducer and used to evaluate the induction of laccase on solid-state fermentation (SSF), using sugarcane bagasse (SCB) as substrate, in an attempt to reach selective delignification. We found that laccase induction occurred in SSF, and a slight inhibition of cellulase production was observed when CA was added to the substrate; also, a transformation of SCB under SSF was followed by the ¹³C cross polarization magic angle spinning (CPMAS) solid-state nuclear magnetic resonance (NMR). Results showed that P. sanguineus can selectively delignify SCB, decreasing aromatic C compounds by 32.67 % in 16 days; O-alkyl C region (polysaccharides) was degraded less than 2 %; delignification values were not correlated with laccase activities. Cellulose-crystallinity index was increased by 27.24 % in absence of CA and 15.94 % when 0.01 mM of CA was added to SCB; this dye also inhibits the production of fungal biomass in SSF (measured as alkyl C gain). We conclude that CA is a good inducer of laccase in liquid media, and that P. sanguineus is a fungus with high potential for biomass delignification.

     

Production and purification of 2-phenylethanol by Saccharomyces cerevisiae using tobacco waste extract as a substrate

2-phenylethanol (2-PE), which is extracted naturally from plant or biotechnology processing, is widely used in the food and cosmetics industries. Due to the high cost of 2-PE production, the valorization of waste carbon to produce 2-PE has gained increasing attention. Here, 2-PE was produced by Saccharomyces cerevisiae using tobacco waste extract (TWE) as the substrate. Considering the toxicity of nicotine and its inhibition of 2-PE, the tolerance of S. cerevisiae was first evaluated. The results suggested that the production of 2-PE by S. cerevisiae in TWEs could be carried out at 2·0 mg ml⁻¹ nicotine concentrations and may be inhibited by 1·0 mg ml⁻¹ 2-PE. Thus, the compounds in the TWEs prepared at different temperatures were detected, and the results revealed that the TWEs prepared at 140°C contained 2·18 mg ml⁻¹ of nicotine, had total sugar concentrations of 26·8 mg ml⁻¹ and were suitable for 2-PE production. Due to feedback regulation, the 2-PE production was only 1·11 mg ml⁻¹, and the remaining glucose concentration remained at 13·78 mg ml⁻¹, which indicated insufficient glucose utilization. Then, in situ product recovery was further implemented to remove this inhibition; the glucose utilization (the remaining concentration decreased to 3·64 mg ml⁻¹) increased, and the 2-PE production increased to 1·65 mg ml⁻¹. The 2-PE produced in the fermentation broth was first isolated by elution from the resin with 75% ethanol and then by removing the impurities with 2·5% activated charcoal, and pure 2-PE was identified by gas chromatography mass spectrometry. The results of this study suggest that TWE could be an alternative carbon source for 2-PE production. This could provide an outlet tobacco waste as well as reducing the price of natural 2-PE, although more strategies need to be explored to improve the production yield of 2-PE by using TWE.     

Optimization studies to develop a low-cost medium for production of the lipases of Rhizopus microsporus by solid-state fermentation and scale-up of the process to a pilot packed-bed bioreactor

A low-cost lipase preparation is required for enzymatic biodiesel synthesis. One possibility is to produce the lipase in solid-state fermentation (SSF) and then add the fermented solids (FS) directly to the reaction medium for biodiesel synthesis. In the current work, we scaled up the production of FS containing the lipases of Rhizopus microsporus. Initial experiments in flasks led to a low-cost medium containing wheat bran and sugarcane bagasse (50:50 w/w, dry basis), supplemented only with urea. We used this medium to scale-up production of FS, from 10 g in a laboratory column bioreactor to 15 kg in a pilot packed-bed bioreactor. This is the largest scale yet reported for lipase production in SSF. During scale-up, the hydrolytic activity of the FS decreased 57%: from 265 U g⁻¹ at 18 h in the laboratory bioreactor to 113 U g⁻¹ at 20 h in the pilot bioreactor. However, the esterification activity decreased by only 14%: from 12.1 U g⁻¹ to 10.4 U g⁻¹. When the FS produced in the laboratory and pilot bioreactors were dried and added directly to a solvent-free reaction medium to catalyze the esterification of oleic acid with ethanol, both gave the same ester content, 69% in 48 h.     

Production of fructosyl transferase by <Emphasis Type="Italic">Aspergillus oryzae</Emphasis> CFR 202 in solid-state fermentation using agricultural by-products

Fructosyl transferase (FTase) production by Aspergillus oryzae CFR 202 was carried out by solid-state fermentation (SSF), using various agricultural by-products like cereal bran, corn products, sugarcane bagasse,cassava bagasse (tippi) and by-products of coffee and tea processing. The FTase produced was used for the production of fructo-oligosaccharides (FOS), using 60% sucrose as substrate. Among the cereal bran used, rice bran and wheat bran were good substrates for FTase production by A. oryzae CFR 202. Among the various corn products used, corn germ supported maximum FTase production, whereas among the by-products of coffee and tea processing used, spent coffee and spent tea were good substrates, with supplementation of yeast extract and complete synthetic media. FTase had maximum activity at 60°C and pH 6.0. FTase was stable up to 40°C and in the pH range 5.0–7.0. Maximum FOS production was obtained with FTase after 8 h of reaction with 60% sucrose. FTase produced by SSF using wheat bran was purified 107-fold by ammonium sulphate precipitation (30–80%), DEAE cellulose chromatography and Sephadex G-200 chromatography. The molecular mass of the purified FTase was 116.3 kDa by SDS-PAGE. This study indicates the potential for the use of agricultural by-products for the efficient production of FTase enzyme by A. oryzae CFR 202 in SSF, thereby resulting in value addition of those by-products.     

Integrated Strategies to Enhance Cellulolytic Enzyme Production Using an Instrumented Bioreactor for Solid-State Fermentation of Sugarcane Bagasse

The conversion of agro-industrial residues, such as sugarcane bagasse, into high-value products and renewable energy, within the biorefinery concept, is a potential alternative towards the sustainable management of these resources. This work evaluates the production of cellulolytic enzymes by a selected strain of Aspergillus niger cultivated in sugarcane bagasse under solid-state fermentation using an instrumented lab-scale bioreactor. The effects of environmental factors including the type of substrate and medium composition, as well as the operational conditions (air flow rate, inlet air relative humidity, and initial substrate moisture content) on the production of the enzymatic complex were evaluated using statistical design tools. Significant increases in FPase, endoglucanase, and xylanase activities were achieved under the optimized conditions predicted by the models, with values of 0.88, 21.77, and 143.85 IU/g of dry solid substrate, respectively, representing around ten-, four-, and twofold increases compared to the activities obtained under the initial growth conditions. This demonstrates the importance of evaluating environmental and operational criteria in order to achieve efficient enzyme production. The crude enzymatic extract obtained under optimized conditions was employed for enzymatic hydrolysis of pretreated sugarcane bagasse. Approximately 13 % of total reducing sugars, and a glucose concentration of 2.54 g/L, were obtained after 22 h of hydrolysis of steam exploded sugarcane bagasse, indicating that the enzymatic cocktail produced has good potential for use in the conversion of biomass.     

Extracellular L-asparaginase production in solid-state fermentation by using sugarcane bagasse as support material

Abstract

L-asparaginase is an important enzyme used in the pharmaceutical and food industry, which can be produced by different microorganisms using low cost feedstocks. In this work, sugarcane bagasse (SCB) was used as support for enzyme production in solid-state fermentation (SSF) by A. terreus. Initially, the influence of the variables carbon and nitrogen sources on the enzyme production was studied following an experimental design carried out in Erlenmeyer flasks. Statistical analysis indicated the use of 0.54% of starch, 0% of maltose, 0.44% of asparagine, and 1.14% of glutamine in the medium, resulting in enzyme activity per volume of produced extract of 120.723?U/L. Then, these conditions were applied in a horizontal column reactor filled with SCB, producing 105.3?U/L of enzyme activity. Therefore, the potential of extracellular L-asparaginase enzyme production in the column reactor using sugarcane bagasse as support was demonstrated and it represents a system that can favor large scale production.     

Ethanol production in simultaneous saccharification and fermentation of cellulose with temperature profiling

The effects of temperature on enzymatic saccharification of cellulose and simulataneous saccharification and fermentation (SSF) were investigated with 100 g·l⁻¹ Solka Floc, 5g·l⁻¹Trichoderma reesei cellulase, and Zymomonas mobilis ATCC 29191. The following results were obtained: 1) Ethanol fermentation under glucose dificient conditions can proceed for more than 100 h at 30°C but gradually ceases after 50 h of operation at 40°C. 2) Equivalent glucose yield based on cellulose for SSF operated at its optimum temperature (37°C) is higher than that for enzymatic saccharification of cellulose at the same temperature by 32%. However, the same equivalent glucose yields were obtained for both processes if they were operated at their respective optimum temperature. 3) SSF with temperature cycling increased the ethanol productivity but gave similar ethanol yield to SSF at 37°C. 4) SSF with temperature profiling gave an ethanol yield of 0.32 g·g⁻¹ and cellulose use of 0.86 g·g⁻¹ which were increased by 39% and 34% over SSF with temperature cycling and at 37°C.     

Effect of inorganic salt stress on the thermotolerance and ethanol production at high temperature of <Emphasis Type="Italic">Pichia kudriavzevii</Emphasis>

Application of cross-protection is expected to improve the thermotolerance of yeasts to enhance their ethanol production at high temperature. In this study, the effects of eight kinds of inorganic salts on the thermotolerance and ethanol production at high temperature in Pichia kudriavzevii were investigated. P. kudriavzevii showed strong thermotolerance and the ability to produce ethanol at high temperature, and higher ethanol production of P. kudriavzevii was observed at high temperature (37–42 °C) compared with that at 30 °C. Inorganic salt stresses induced obvious cross-protection of thermotolerance in P. kudriavzevii. The presence of 0.1 mol/L KNO₃ or Na₂SO₄ or 0.2 mol/L NaCl, KCl, NaNO₃, K₂SO₄ or MgCl₂ increased the yeast biomass in YEPD medium at 44 °C to 2.72–3.46 g/L, obviously higher than that in the absence of salt stress (2.17 g/L). The addition of NaCl, KCl, NaNO₃, KNO₃, Na₂SO₄, K₂SO₄, CaCl₂ and MgCl₂ significantly increased the ethanol production of P. kudriavzevii in YEPD fermentation medium at 44 °C by 37–58%. KCl and MgCl₂ exhibited the best performance on improving the thermotolerance and ethanol production, respectively, of P. kudriavzevii. A highly significant correlation (P?<?0.01) was obtained among ethanol production, biomass and glucose consumption, suggesting the important role of thermotolerance and glucose consumption in enhanced ethanol production. The combination of NaCl, KCl and MgCl₂ had a synergistic effect on the improvement of thermotolerance and ethanol production at high temperature in P. kudriavzevii. This study provides some important clues for improving ethanol production of thermotolerant yeasts at high temperature.     

Efficient conversion of high concentration of glycerol to Monacolin K by solid-state fermentation of Monascus purpureus using bagasse as carrier

High concentration of glycerol was used as the sole carbon source for efficient production of Monacolin K (MK) by solid-state fermentation (SSF) of Monascus purpureus 9901 using agricultural residue (bagasse), as an inert carrier. A comparative study showed that MK production in SSF was about 5.5 times higher than that of submerged fermentation when 26 % of glycerol was used, which may be due to the formation of glycerol concentration gradients in the inert carrier and less catabolite repression in SSF. For enhancement of MK yield in SSF, the effects of different influential variables, such as glycerol concentration, nitrogen source and its concentration, initial moisture content, inoculum size and particle size of bagasse, were systematically examined. All the factors mentioned above had an effect on the MK production in SSF to some extent. The maximal yield of MK (12.9 mg/g) was achieved with 26 % glycerol, 5 % soybean meal, 51 % initial moisture content, 20 % inoculum size and 1 mm particle size of bagasse. The results in this study may expand our understanding on the application of SSF using agricultural residue as carrier for production of useful microbial metabolites, especially the efficient conversion of high concentration of glycerol to MK by Monascus purpureus.     

A note on utilization of bagasse for the production of proteinaceous cattle feed

The basidiomycete cultures, Polyporus strains BH₁ and BW₁, were cultivated on whole bagasse substrate in a solid-state system. The cultures degraded cellulose, hemicellulose and lignin contents of bagasse, increasing the digestibility and protein content of the product, to be utilized as cattle feed. Untreated and unwashed bagasse proved a good substrate.     

Comparing submerged and solid-state fermentation of agro-industrial residues for the production and characterization of lipase by Trichoderma harzianum

Lipase production by Trichoderma harzianum was evaluated in submerged fermentation (SF) and solid-state fermentation (SSF) using a variety of agro-industrial residues. Cultures in SF showed the highest activity (1.4 U/mL) in medium containing 0.5 % (w/v) yeast extract, 1 % (v/v) olive oil and 2.5 C:N ratio. This paper is the first to report lipase production by T. harzianum in SSF. A 1:2 mixture of castor oil cake and sugarcane bagasse supplemented with 1 % (v/w) olive oil showed the best results among the cultures in SSF (4 U/g ds). Lipolytic activity was stable in a slightly acidic to neutral pH, maintaining 50 % activity after 30 min at 50 °C. Eighty percent of the activity remained after 1 h in 25 % (v/v) methanol, ethanol, isopropanol or acetone. Activity was observed with vegetable oils (olive, soybean, corn and sunflower) and long-chain triacylglycerols (triolein), confirming the presence of a true lipase. The results of this study are promising because they demonstrate an enzyme with interesting properties for application in catalysis produced by fermentation at low cost.     

Influence of planting geometry on photosynthetically active radiation interception and dry matter production relationships in pearl millet

A field experiment was conducted at the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) Center, Patancheru, India to study photosynthetically active radiation (PAR) interception and dry matter production relationships in pearl millet (Pennisetum americanum (L.) Leeke). Two pearl millet genotypes, BJ 104 (G₁) and ICH 226 (G₂) were sown at three planting geometries obtained by using combinations of row and plant spacings (S₁: 37·5 cm × 26·6 cm; S₂: 75·0 cm × 13·3 cm; S₃: 150·0 cm × 6·6 cm) such that plant population was constant at 100 000 ha⁻¹ in all treatments. Cumulative intercepted PAR was maximum (330 MJ m⁻²) in G₂S₂ and minimum (268 MJ m⁻²) in G₁S₃. Conversion efficiency values ranged from 1·87 g MJ⁻¹ in G₁S₂ to 2·32 g MJ⁻¹ in G₂S₃. Final above-ground dry matter followed the pattern of cumulative intercepted PAR and maximum dry matter (7·22 Mg ha⁻¹) was produced by G₂S₂ while G₁S₃ produced minimum dry matter (4·97 Mg ha⁻¹).     

Simplification of the Biomass to Ethanol Conversion Process by Using the Whole Medium of Filamentous Fungi Cultivated Under Solid-State Fermentation

A novel simplified configuration is proposed for the conversion of biomass to ethanol using whole medium enzymatic cocktails (WM) and enzymatic extracts (EE) from different filamentous fungi (Trichoderma reesei, Aspergillus niger, and Aspergillus oryzae) cultivated under solid-state fermentation (SSF) for the hydrolysis of steam-exploded sugarcane bagasse (SESB). The hydrolyzed material derived from the saccharification of SESB using the combinations A. niger WM + T. reesei EE, A. oryzae WM + A. niger EE, and A. niger EE + T. reesei WM resulted in the best biomass conversion yields (66, 65, and 64 % of the theoretical reducing sugar yields, respectively). The best ethanol production (84 % of the theoretical yield) was obtained using the material hydrolyzed by a combination of A. oryzae WM + A. niger EE. The enzymatic conversion of SESB using on-site produced enzymes from the whole SSF cultivation medium, followed by an ethanol production step, is a potential configuration for the biomass to ethanol conversion process. This novel simplified configuration would enable the use of a single reactor system, avoiding the need for additional separation steps.