Printer Friendly

Citric waste saccharification under different chemical treatments/Sacarificacao de residuo citrico sob diferentes tratamentos quimicos.

Introduction

Biofuels play an important role in reducing changes in global climate. Their impact depends upon several aspects related to novel technologies, legal restrictions, international trade, land use as well as the choice of raw materials and management techniques (WORLDWATCH INSTITUTE, 2007). During the last two decades, second generation ethanol has been proposed as an alternative for biofuel production, though hydrolysis and fermentation of lignocellulosic materials have been known and implemented since the late nineteenth century. Several studies have been executed particularly in the USA and in Europe, albeit still within laboratory scale, and aim at an efficient biofuel capable of being produced worldwide. Moreover, all biomass wastes derived from agribusiness, agro-industry residues and urban waste have high lignocellulosic contents (MACEDO et al., 2008).

Biomass production costs in Brazil are considered the lowest in the world, with further possibilities in achieving more promising results. The production of lignocellulosic ethanol is expected to increase up to 50% ethanol production without the need of expanding the area of current plantations (SILVA, 2012).

With approximately 35% of global production, estimated at 47,010 thousand tons, Brazil is the leading orange producer worldwide, followed by USA, China, India, Mexico, Egypt and Spain. The Brazilian 2013 crop produced about 16.3 million tons of oranges with an expected increase of 1% in 2014 (IBGE, 2014). As a result of this large-scale orange processing, great amounts of waste are generated since orange bagasse corresponds to 50% of its fresh weight. On a dry basis, orange biomass features nearly 16% hemicellulose, 28% cellulose and 9% lignin (RETORE et al., 2010), denoting an alternative for the production of cellulose or second generation ethanol (LENNARTSSON et al., 2012).

The technology for obtaining bioethanol from lignocellulosic materials comprises the hydrolysis of biomass polysaccharides into fermentable sugars and further fermentation, standing out as a feasible energetic alternative to meet global demands. Bellido et al. (2011) pointed out five unit operations required for an efficient conversion of lignocellulosic biomass into ethanol: (1) biomass size reduction to increase surface area and uniformity; (2) pretreatment to break lignin and hemicellulose structures, reducing cellulose crystallinity while increasing biomass porosity; (3) enzymatic hydrolysis to convert polymeric sugars into monomeric ones; (4) fermentation, to produce ethanol from monomeric sugars; (5) ethanol recovery by distillation or any other separation technique.

The lignocellulosic biomass is composed of cellulose (a polysaccharide formed by glucose molecules linked by P-1.4-glycoside bonds) chains joined by hydrogen interactions. These long cellulose fibers are coated with hemicelluloses which are branched polysaccharides mainly consisting of D-xylose and small amounts of L-arabinose, D-glucose, D-mannose, D-galactose, glucuronic acid, mannuronic acid and lignin (WYMAN et al., 2005).

The main pretreatment technologies are chemical pretreatments, including acid, alkaline and oxidative treatments. Most pretreatments differ in chemical structure and in the mechanisms of cell wall chemical and structural modification, which in turn leads to improved enzyme accessibility and increased yields (OGEDA; PETRI, 2010). Table 1 presents the activities of different pretreatment types as to lignocellulosic structure chemical and conformational modifications.

Pretreatment with diluted sulfuric acid thoroughly hydrolyzes the hemicellulose fraction to the medium which, depending on the acid concentrations employed, also releases cellulose to a greater or lesser extent, and other components such as pectin and water-soluble proteins (CORTEZ, 2010). Contrastingly, milder operating conditions (temperature and pressure) are employed in alkaline processes whose main effect is the removal of lignin from the biomass, enhancing higher fiber reactivity (MOSIER et al., 2005; ROCHA et al., 2009). The alkali, usually soda, tends to cause the swelling of the biomass decreasing cellulose crystallinity and increasing surface area and porosity (PITARELO et al., 2012). The hydrothermal pretreatment consists in the combination of water and biomass for 15 min. at 230[degrees]C where between 40 and 60% of total biomass is dissolved in water, generating 4-22% cellulose, 30-60% lignin, coupled to complete hemicellulose removal (MOSIER et al., 2005).

Some authors have evaluated orange residue pretreatment by using diluted acid as a catalyst to saccharify hemicellulose from the biomass, with low acid (< 1%) and high biomass concentrations. Vaccarino et al. (1989) employed diluted sulfuric acid and [C.sub.biomass] ranging from 7.4 to 18.5% at 100[degrees]C for a long heating time (1.5 hour). Talebnia et al. (2008) used [C.sub.acid] between 0 and 1%, temperatures from 100 to 132[degrees]C, [C.sub.biomass] from 2 to 18%, and heating times ranging from 5 to 25 min. in citrus residues, with maximum saccharification of 45%. Miller et al. (2012) studied the saccharification of orange waste using 6% sulfuric acid for 15 to 120 min. and employing [C.sub.biomass] of 10% biomass, with a yield of about 30% of total reducing sugars. Employing a hydrothermal process with orange residue, Pitarelo et al. (2012) applied temperatures between 195 and 210[degrees]C and short heating times (4, 6, and 8 min.) and reported that the degradation of sugars in the liquor was enhanced by increased temperatures. Grohann et al. (1995) used hydrothermal process at milder temperatures (100-140[degrees]C) and Cbiobass of 1% and obtained 55-65% saccharifications.

Current analysis evaluates the severity of acidic and alkaline pretreatments on liquor saccharification, solubilization of solid fraction and mass yield, by studying the binomial time and [C.sub.acid] or [C.sub.alkali] at different solid concentrations (low to moderate, 1 to 9%) and high catalyst concentrations. A hydrothermal pretreatment was performed under the same conditions of acidic and alkaline processes to investigate the relative increase in selectivity by using the above-mentioned catalysts.

Material and methods

The methodology consisted of waste preparation, physicochemical characterization, and acidic, alkaline or hydrothermal pretreatment, resulting in pretreated biomass and liquor.

Waste, obtained after juice/pulp extraction at the processing plant (COOPLAL, in Santana do Mundau AL Brazil), was collected in plastic bags, ice-cold stored and taken to the laboratory where it was thawed, sanitized with a solution containing 100 ppm of sodium hypochlorite for 15 min., kiln-dried at 55[degrees]C until constant weight was reached, crushed in Wyllie mill with a 30 mesh sieve, and stored airtight in plastic bottles at room temperature.

Moisture, ash, protein, lipid, fiber, sugar and pectin contents were determined in waste samples, following analytical procedures by the Adolfo Lutz Institute (IAL, 2005) and AO AC (2002). The percentage of total carbohydrates was calculated by the difference of the analyses described above. Reducing (RS) and total reducing (TRS) sugars were determined by colorimetry (MILLER, 1959) with 3.5-dinitrosalicylic (DNS) acid. For the determination of TRS, samples were treated with [H.sub.2]S[O.sub.4] 1.5 M in boiling water for 20 min. with occasional stirring to hydrolyze polysaccharides and other non-reducing sugars.

Three pretreatments were performed: (1) acidic, with diluted sulfuric acid ([H.sub.2]S[O.sub.4]) solutions; (2) alkaline, with sodium hydroxide (NaOH) solutions; and (3) hydrothermal, with distilled water only. Response surface methodology (RSM) was used to optimize the pretreatment conditions. For the acidic and alkaline pretreatments, the independent variables were time, solution concentration ([C.sub.acid] or [C.sub.alkali]), and biomass concentration ([C.sub.biomass]), whereas for hydrothermal pretreatment, the variables were time and [C.sub.biomass] only. Data were treated with Statistica 7.0, and the response variables were mass yield (MY), TRS content released in the pretreatment liquor, and total soluble solids (TSS) in the liquor.

Response surfaces were built from significant variables. Analysis of variance (ANOVA) was employed to validate the model proposed by Statistica, according to Equation 1.

Y = [[beta].sub.0] + [[SIGMA].sup.k.sub.i=1] [[beta].sub.ij][X.sub.i][X.sub.j] + [[SIGMA].sup.k.sub.j=1][[beta].sub.j][X.sup.2.sub.j] (1)

where:

Y is the response variable; [[beta].sub.0] is a constant; [[beta].sub.i], [[beta].sub.j] and [[beta].sub.ij] are the linear, quadratic and interaction coefficients, respectively.

The central composite design (CCD) was used to acquire data to fit the above equation. In acidic and alkaline pretreatments, a [2.sup.3] full factorial design was used, whereas a [2.sup.2] full factorial design was employed in the hydrothermal pretreatment, both of which including three replicates at the central point, resulting in eleven and nine experiments, respectively, to investigate the selected variables.

The environmental conditions were 121[degrees]C and 1 atm, monitored in autoclave. After pretreatment in the autoclave reactor, samples were filtered and their liquors were analyzed as to TRS by DNS method (MILLER, 1959), according to Equation 2. The solid fraction was dried at 37[degrees]C for 24 hours and then weighed.

%TRS = 100. C (g.[L.sup.-1]). V (L)/Biomass (g) (2)

where:

C (g [L.sup.-1]) is the TRS concentration obtained from a glucose standard curve;

Y (L) is the volume of the extracted liquor;

Biomass (g) is the waste mass.

Equation 3 gives the calculation of mass yield (MY) which considers the biomass's initial and final weights (before and after the pretreatment, respectively), whereas Equation 4 considers solubilization by a relationship between TSS ([degrees]Brix) and waste mass.

%MY = 100. [m.sub.final](g)/[m.sub.initial](g) (3)

TSS = [degrees]Brix/Biomass(g) (4)

Results and discussion

The waste's drying process took 18 hours, with approximately 85% moisture loss. The physicochemical characterization indicated high carbohydrate contents (Table 2) and showed itself compatible with the characterizations reported elsewhere, suggesting that the residue is a potential source for second generation ethanol production.

Table 3 gives results of the acidic pretreatment. It may be observed that sugar contents released to the liquid fraction after pretreatment remained between 27 and 62%, underscoring the great influence of the studied variables on the saccharification process. In soluble solid and TRS (Total Reducing Sugars) release profiles, due to material hydrolysis as a result of heating in acidic medium, it may be observed that, regardless of heating time, low biomass and high acid concentrations resulted in high quantities of dissolved substances when compared to the biomass used in each experiment.

The above suggests that, besides pectin and hemicellulose fractions, a portion of another carbohydrate, most likely cellulose, was also released during the process. This occurred because the experiment with a greater saccharification degree, featuring the shorter pretreatment (15 min.), the lowest biomass concentration (1 g 100 m[L.sup.-1]) and the highest acid concentration (5 mL 100 m[L.sup.-1]), resulted in a saccharification degree of 62% (70% of the total theoretical carbohydrates).

The acidic pretreatment showed that [C.sub.biomass] negatively affected sugar solubilization, probably due to the low solid/liquid ratio interfering with structure breakdown. In their study on the pretreatment of orange peel in diluted sulfuric acid (0.2 to 0.6%) with [C.sub.biomass] between 7.4 and 18.5%, at 100[degrees]C for 1.5 hour, Vaccarino et al. (1989) reported that [C.sub.biomass] increase caused higher mass yield (from 39 to 62%). This behavior was similar to the observations in current assay, although [C.sub.biomass] was not significant at 95% as to mass yield in the studied range (1-9%). It has also been observed that TRS released to the liquor varied between 21.3 and 45.8%, which are the highest rates obtained at higher [C.sub.acid] and lower [C.sub.biomass], according to current results.

[C.sub.acid] positively influenced waste matrix solubilization at higher acid concentrations, resulting in lower mass yield and indicating a major biomass solubilization. Grohann et al. (1995) used sulfuric acid at concentrations 0.06 and 0.5%, temperatures between 100 and 140[degrees]C, and pretreatments for 10-40 min. and achieved mass yields between 18 and 30%, with lower yields being obtained for longer times, as observed in current study. At 120[degrees]C and heating times between 10 and 40 min., there was a TRS dissimilation of 30 to 37% and of 11% for [C.sub.acid] of 0.5 and 0.06%, respectively. These rates were lower than those found in the current analysis, probably due to lower [C.sub.acid] employed. The above indicates the positive influence of acid for sugar dissimilation, as shown in the Pareto chart (Figure 1), even though its influence was not significant, at 95%, within the studied range, unlike its interaction with time.

However, [C.sub.acid] negatively affected sugar solubilization since, as previously enhanced, at a certain time there was a severity that led to sugar degradation, provided that mass yield was not significantly influenced by the assessed variables, i.e., p < 0.05, between 30 and 40%. The R2 coefficient shows that regression models adjusted for TRS (0.9953), mass yield (0.9965), and biomass solubilization (0.8906) were suitable.

Reduced amount of sugars with increasing time and higher catalyst concentration may be due to degradation of these compounds. In fact, sugar degradation is influenced by acids whilst high temperatures and long exposure times also generated compounds capable of inhibiting further fermentation, including hydroxymethylfurfural (HMF) and furfural (MOSIER et al., 2005; CARA et al., 2008).

Approximately 30% of the cellulose was saccharified at 170[degrees]C when olive wood was used (CARA et al., 2008). In fact, under severe pretreatment conditions, part of the cellulose was solubilized in the pretreatment liquor, i.e., some cellulose was actually lost prior to the enzymatic hydrolysis process. Additionally, hemicellulose recovery in the liquor remarkably dropped to 5% with longer times and higher temperatures, which also suggesting pentose degradation.

In the studied range (15 to 120 min.), time played a significant role in biomass saccharification because its interaction with [C.sub.acid] was significantly influential. Saccharification of about 30% was obtained in the liquor of orange waste after hydrolysis with diluted sulfuric acid at 122[degrees]C, using [C.sub.acid] of 6 mL 100 [mL.sup.-1] and [C.sub.biomass] of 10 g 100 [mL.sup.-1], without much variation for the pretreatment times 15 to 120 min. (MILLER et al., 2012). This yield was similar to that in current assay for 9% of [C.sub.biomass] and 15 min and showed that at higher [C.sub.biomass] (~ 10%), 15 min. was the most suitable pretreatment time for saccharification.

Talebnia et al. (2008) studied orange bagasse hydrolysis in diluted sulfuric acid with [C.sub.biomass] between 2 and 18% (w [v.sup.-1]), [C.sub.acid] from 0 to 1% (w [v.sup.-1]), temperatures at 100, 108, 116, 124, and 132[degrees]C, and heating times in autoclave between 5 and 25 min., and observed that the best saccharification (approximately 45% of saccharification in TRS) occurred after the use of 116[degrees]C, 0.5% sulfuric acid, 6% [C.sub.biomass], and 15 min. heating, suggesting mild heating (116[degrees]C) and [C.sub.acid] 0.5% as optimal parameters. The authors also reported that HMF formation and saccharified sugar degradation were first detected at the highest temperatures ([greater than or equal to] 124[degrees]C), heating times (>15 min.), [C.sub.acid] ([greater than or equal to] 0.75%) and [C.sub.biomass] of 4%.

The models obtained for TRS and TSS ([[degrees]Brix.g.sub.biomass.sup.-1]) are given in Equations 5 and 6, respectively. The MY model was not significant and might have been affected by the acidic pretreatment conditions.

[%TRS.sub.acid] = 64.153 - 0.7441.time + 0.0066. [time.sup.2] - 3.5086. [C.sub.biomass] - 0.0372. time. [C.sub.acid] (5)

[TSS.sub.acid] = 1.317 - 0.0486.time + 0.0004.[time.sup.2] + 1.2792. [C.sub.acid] - [degrees].0465. [C.sub.biomass] - 0.1252. [C.sub.acid].[C.sub.biomass] (6)

In the alkaline pretreatment, lower mass yields (about 30%) were obtained, except for Experiments 3 and 4, due to the fact that the two experiments generated a gum and impaired pretreatment efficiency because of too much biomass (9%) for pretreatment time. The results are presented in Table 4.

The removal of lignin and hemicellulose fractions is a characteristic of alkaline treatments (SUN; CHENG, 2002; MOSIER et al., 2005) and maybe only a small fraction of these as well as a cellulose fraction may have remained. In current assay, high TSS and, consequently, higher solubilization of fractions were obtained, as well as lower saccharification when compared to acidic pretreatment. It is worth mentioning that sugar dissimilation alone in the liquor during the pretreatment does not evaluate the process efficiency because it could be affected by the saccharification of the hexoses present, mainly in the cellulosic fraction, indicating the need of further enzymatic hydrolysis or characterization of the resulting solid fraction.

Studies with banana stem showed that, for enzymatic hydrolysis, the alkaline pretreatment with NaOH 1% (m [v.sup.-1]) for 1 h and at 100[degrees]C was more efficient, reaching glucose yields of about 61%. If compared with that of the acidic pretreatment, such a high yield has to do with delignification and cellulose loss, the latter being around 30% for the acidic pretreatment and only 1.2% for the alkaline one (GONCALVES FILHO et al., 2013). Approximately 80% of lignin was removed after delignification of previously steam-exploded sugarcane straw with 1% NaOH 1%, for 1 hour, at 100[degrees]C (OLIVEIRA et al., 2013).

Figure 2 presents the Pareto charts for each response variable. Lower [C.sub.biomass] and [C.sub.alkali] influenced sugar extraction in the liquor, where 9 g of biomass and 15 min. of reaction were not enough to make the suspension liquid; a gelatinous mixture was yielded. As to mass yield, lower [C.sub.alkali] and time favored the solubilization of the lignocellulosic fraction, suggesting [C.sub.biomass] 5% and 67.5 min. as the best conditions. One hour at 100[degrees]C has been reported as effective for the delignification of biomass from agro-industrial residues (GONCALVES FILHO et al., 2013; OLIVEIRA et al., 2013).

In the case of TSS, the interaction between [C.sub.alkali] and [C.sub.biomass] appears to have played an important role in the solubilization of lignocellulosic matrix substances. This reflects the susceptibility of the lignocellulosic matrix, mainly lignin and hemicellulose, to NaOH (SUN; CHENG, 2002; MOSIER et al., 2005). In the treatment of agro-industrial residues, 1% of NaOH at 100[degrees]C may be effective in the preparation of biomass to enzymatic hydrolysis (GONCALVES FILHO et al., 2013; OLIVEIRA et al., 2013), revealing the influence of almost all the variables of the experimental design and their interactions in mass yield.

For pretreating biomass of fruit residues, several authors have considered the use of alkaline pretreatment for the removal and/or breakdown of lignocellulosic structures after using diluted sulfuric acid to remove hemicellulose at moderate temperature (100[degrees]C). The literature reports a relative efficiency in the association of these two processes. In an acidic pretreatment to delignify cashew bagasse, approximately 75% of mass yield was achieved after pretreatment with low sulfuric acid concentration (0.8 mol [L.sup.-1]) and [C.sub.alkali] of about 4%, for 1 hour and at 100[degrees]C (ROCHA et al., 2009). When cashew bagasse was pretreated with 0.5 to 3.5% of acid and 1.5% of alkali, glucose conversions of up to 60% were attained (ROCHA et al., 2014). Although the combination of these processes might be efficient in agro-industrial waste treatment, studies on cost and feasibility must be undertaken to this end.

The use of alkaline solutions has the drawback of their neutralization and disposal, coupled to making the liquor useless due to numerous lignin-derived inhibitors (JORDAN et al., 2012). This would probably be the function of the process with diluted acid: initial removal of hemicelluloses, favoring lignin breakage by NaOH and, consequently, improving the enzymatic process.

ANOVA parameters indicated R2 coefficients related to the response variations better than those adjusted to the acidic pretreatment, especially to mass yield (0.9986). The models obtained for TRS, biomass solubilization and mass yield are given in Equations 7 to 9, respectively.

[%TRS.sub.alkali] = 20.5866 + 0.0913. time - 2.5327.[C.sub.biomass] + 0.9172. [C.sub.alkali].[C.sub.biomass] (7)

[TSS.sub.alkali] = 1.7728 + 0.1451. [C.sub.alkali] - 0.2574. [C.sub.biomass] + 0.3031. [C.sub.alkali].[C.sub.biomass] (8)

[%MY.sub.alkali] = 33.0219 - 0.5729.time + 0.0036.[time.sup.2] + 0.1969. [C.sup.alkaii] + 7.6843.[C.sub.biomass] + 0.044.time.[C.sub.alkali] - 3.0269. [C.sub.alkali].[C.sub.biomass] (9)

In the case of hydrothermal pretreatment, divergences were noted in the results (Table 5), chiefly regarding TRS and TSS contents released by the biomass in the liquor, with rates between 3 and 37 and between 0.30 and 0.95, respectively. In fact, they are the highest rates obtained when lower biomass concentrations were used.

The Pareto charts in Figure 3 demonstrate that, among the studied factors, the lowest [C.sub.biomass] was significant in both responses and suggested that high biomass causes lower self-catalytic action and lessens the biomass water/surface ratio or sugar degradation due to prolonged exposure.

Pitarelo et al. (2012) hydrothermally processed bagasse and sugar cane straw for 4, 6, and 8 min., at temperatures of 195, 202.5, and 210[degrees]C, and with moisture contents of 8, 33, and 50%. Higher residence times in the reactor, as well as elevated temperatures, led to greater losses in the pretreatment process. These losses may be attributed to the prolonged exposure of the material to high temperatures and to the further increase in the degradation rates of the carbohydrates present in the bagasse (decomposition).

Mass yield ranged between 45 and 60%, with increased [C.sub.biomass] (%) and reduced time being significant variables. In the hydrothermal treatment of 1% citrus waste, temperatures of 100, 120, and 140[degrees]C and reaction times between 5 and 40 min., mass yields ranging from 55 to 65% were obtained at 120[degrees]C for 40 min., with the longest time resulting in the lowest mass yield (GROHANN et al., 1995).

It is likely that the temperature employed was not enough to breakdown the lignocellulosic structure, since in the absence of chemical catalysts (acid or alkali), the catalytic action was reduced. Many authors reported temperatures between 175 and 225[degrees]C as the most effective for this pretreatment (SUN; CHENG, 2002; MOSIER et al., 2005). Given the low saccharification in the liquor, enzymatic hydrolysis tests may confirm the efficiency of this process. However, biomass reactivity was greater in the previous pretreatments.

The models obtained for TRS, biomass solubilization and mass yield are given in Equations 10 to 12, respectively.

[%TRS.sub.hydrothermal] = 50.6145 - 11.8320.[C.sub.biomass] + 0.7785.[C.sub.biomass.sup.2] (10)

[TSS.sub.hydrothermal] = 1.0326 - 0.1609.[C.sub.biomass] - 0.0082.[C.sub.biomass.sup.2] (11)

[%MY.sub.hydrothermal] = 51.5573 - 0.1425.time + 1.82021.[C.sub.biomass] (12)

Table 6 presents a comparison between the different pretreatments that, albeit distinct, had similar influences on lower [C.sub.biomass] and sugar saccharification, the positive significance for time when alkali or acid were present, and the fact that [C.sub.acid] or [C.sub.alkali] alone was not significant, but only together with other variables.

Based on these data and on the discussions above, it is believed that long reaction times, low acid or alkali concentrations and low biomass concentrations should be used for high saccharification rates. However, the operating costs are high, encouraging the resort to shorter times and higher [C.sub.biomass]. Further studies on enzymatic hydrolysis and composition of the resulting solid fraction must be undertaken to better support these hypotheses. It should be also noted that saccharification is considerably high in the pretreatment where cellulose is probably lost.

Conclusion

Interactions between the variables (time, [C.sub.acid] and [C.sub.biomass]) have been analyzed. The sugar range was much larger and indicated that time and [C.sub.acid] might contribute towards sugar liquor degradation when the most severe conditions are used. The use of higher [C.sub.biomass], lower [C.sub.acid], and shorter times may be effective. In alkaline pretreatment, [C.sub.alkali] did not affect mass yield; however, time affected it negatively, suggesting the use of lower [C.sub.alkali], treatment times between 67.5 and 120 min. and [C.sub.biomass] of 5%. The hydrothermal pretreatment produced the highest mass yields (60%), recommending lower pretreatment times and higher [C.sub.biomass].

Doi: 10.4025/actascitechnol.v37i4.28133

Acknowledgements

The authors would like to thank PPGEQ-UFAL and Capes for their financial support and fellowships.

References

AOAC-Association of Official Analytical Chemists. Official methods of analysis of the association of official analytical chemists. 17th ed. Gaithersburg: William Horwitz, 2002.

BELLIDO, C.; BOLADO, S.; COCA, M.; LUCAS, S.; GONZALEZ-BENITO, G.; GARCIA-CUBERO, M. T. Effect of inhibitors formed during wheat straw pretreatment on ethanol fermentation by Pichia stipitis. Bioresource Technology, v. 102, p. 10868-10874, 2011.

CARA, C.; RUIZ, E.; OLIVA, J. M.; SAEZ, F.; CASTRO, E. Conversion of olive tree biomass into fermentable sugars by dilute acid pretreatment and enzymatic saccharification. Bioresource Technology, v. 99, p. 1869-1876, 2008.

CLEMENTE, E.; FLORES, C. A.; ROSA, F. L. I. C.; OLIVEIRA, M. D. Caracteristicas da farinha de residuo do processamento da laranja. Revista Ciencias Exatas e Naturais, v. 14, n. 2, p. 257-269, 2012.

CORTEZ, L. A. B. Bioetanol de cana-de-acucar: P&D para produtividade e sustentabilidade. Sao Paulo: Blucher, 2010.

GONCALVES FILHO, L. C.; FISCHER, G. A. A.; SELLIN, N.; MARANGONI, C.; SOUZA, O. Hydrolysis of banana tree pseudostem and second-generation ethanol production by Saccharomyces cerevisae. Journal of Environmental Science and Engineering, v. 2, n. 1A, p. 65-69, 2013.

GROHANN, K.; CAMERON, G. R.; BUSLIG, S. B. Fractionation and pretreatment of orange peel by dilute acid hydrolysis. Bioresource Technology, v. 54, p. 129-141, 1995.

IAL-Instituto Adolfo Lutz. Metodos fisico-quimicos para analises de alimentos. 4. ed. Sao Paulo: IAL, 2005. IBGE-Instituto Brasileiro de Geografia e Estatistica. Indicadores agropecuarios. Brasilia: IBGE, 2014.

JORDAN, D. B.; BOWMAN, M. J.; BRAKER, J. D.; DIEN, B. S.; HECTOR, R. E.; LEE, C. C.; MERTENS, J. A.; WAGSCHAW, K. Plant cell walls to ethanol. Biochemical Journal, v. 442, p. 241-252, 2012.

LENNARTSSON, P. R.; YLITERVO, P.; LARSSON, C.; LARS, E.; TAHERSADEH, M. J. Growth tolerance of Zygomycetes Mucor indicus in orange peel hydrolysate without detoxication. Process Biochemistry, v. 47, n. 5, p. 836-842, 2012.

MACEDO, I. C.; SEABRA, I. E. A.; SILVA, E. A. R. Greenhouse gases emissions in the production and use of ethanol from sugarcane in Brazil: "The 2005/2006 averages and a prediction for 2020". Biomass and Bioenergy, v. 32, p. 582-595, 2008.

MILLER, F.; CALDEIRAO, L.; MARINELLI, C. D.; SERGIO, P. Obtencao de acucares fermentesciveis a partir da casca de laranja e bagaco de cana-de-acucar. Analytica, v. 59, p. 2-6, 2012.

MILLER, G. L. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, v. 31, n. 3, p. 426-428, 1959.

MOSIER, N.; WYMAN, C.; DALE, B.; ELANDER, R.; LEE, Y. Y.; HOLTZAPPLE, M.; LADISCH, M. Features of promising technologies for pretreatment of lignocellulosic biomass. Bioresource Technology, v. 96, p. 673-686, 2005.

OGEDA, T. L.; PETRI, D. F. S. Hidrolise enzimatica de biomassa. Quimica Nova, v. 33, n. 7, p. 1549-1558, 2010.

OLIVEIRA, F. M. V.; PINHEIRO, I. O.; SOUTOMAIOR, A. M.; MARTIN, C.; GONCALVES, A. R.; ROCHA, G. J. M. Industrial-scale steam explosion pretreatment of sugarcane straw for enzymatic hydrolysis of cellulose for production of second generation ethanol and value-added products. Bioresource Technology, v. 130, p. 168-173, 2013.

PITARELO, A. P.; SILVA, T. A.; PERALTA-ZAMORA, P. G.; RAMOS, L. P. Efeito do teor de umidade sobre o pre-tratamento a vapor e a hidrolise enzimatica do bagaco de cana-de-acucar. Quimica Nova, v. 35, n. 8, p. 1502-1509, 2012.

RETORE, M.; SILVA, L. P.; TOLEDO, G. S. P.; ARAUJO, I. G. Efeito da fibra de co-produtos agroindustriais e sua avaliacao nutricional para coelhos. Arquivo Brasileiro de Medicina Veterinaria e Zootecnia, v. 62, n. 5, p. 1232-1240, 2010.

ROCHA, A. S.; SILVA, F. L. H.; CONRADO, L. S.; LIMA, F. C. S.; CARVALHO, J. P. D.; SANTOS, S. F. M. Ethanol from cashew apple bagasse by enzymatic hydrolysis. Chemical Engineering Transactions, v. 37, p. 361-366, 2014.

ROCHA, M. V. P.; RODRIGUES, T. H. S.; MACEDO, G. R. M.; GONCALVES, L. R. B. Enzymatic hydrolysis and fermentation of pretreated cashew apple bagasse with alkali and dilute sulfuric acid for bioethanol production. Applied Biochemistry and Biotechnology, v. 155, p. 407-417, 2009.

RUVIARO, L.; NONELLO, D.; ALMEIDA, M. J. Analise sensorial de sobremesa acrescida a farelo de casca e bagaco de laranja entre universitarios de Guarapuava (PR). Revista Solus-Guarapuava, v. 2, n. 2, p. 41-50, 2008.

SILVA, F. V. Panorama e perspectivas do etanol lignocelulosico. Revista Liberato, v. 13, n. 20, p. 43-58, 2012.

SUN, Y.; CHENG, J. Hydrolysis of lignocellulosic materials for ethanol production: a review. Bioresource Technology, v. 83, n. 1, p. 1-11, 2002.

TALEBNIA, F.; POURBAFRANI, M.; LUNDIN, M.; TAHERZADEH, M. J. Optimization study of citrus wastes saccharification by dilute-acid hydrolysis. Bioresources, v. 3, n. 1, p. 108-122, 2008.

VACCARINO, C.; CURTO, L. R.; TRIPODO, M. M.; PATANE, R.; LAGANA, G.; RAGNO, A. SCP from orange peel by fermentation with fungi acid-tread peel. Biological Wastes, v. 30, p. 1-10, 1989.

WORLDWATCH INSTITUTE. Biofuels for Transport. Washington, D.C.: Earthscan, 2007.

WYMAN, C. E.; DECKER, S. R.; HIMMEL, M. E.; BRADY, J. W.; SKOPEC, C. E.; VIIKARI, L. Polysaccharides: structural diversity and functional versatility. New York: Dekker, 2005.

Received on June 11, 2015.

Accepted on July 10, 2015.

License information: This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Carlos Eduardo de Farias Silva (1) *, Georgia Nayane Silva Belo Gois (1), Livia Manuela Oliveira da Silva (1), Renata Maria Rosas Garcia Almeida (1) and Ana Karla de Souza Abud (2)

(1) Centro de Tecnologia, Universidade Federal de Alagoas, Avenida Lourival Melo Mota, s/no., 57072-970, Tabuleiro dos Martins, Maceio, Alagoas, Brazil. (2) Departamento de Tecnologia de Alimentos, Universidade Federal de Sergipe, Sao Cristovao, Sergipe, Brazil. * Author for correspondence. E-mail: eduardo.farias.ufal@gmail.com
Table 1. Effect of various pretreatment methods on the structure
of lignocellulosic biomass.

Pretreatment   Available     Cellulose de     Hemicellulose
                surface    -crystallization      removal
                 area

Diluted acid      (a)             ND               (a)
Alkali            (a)             ND               (b)
Hydrothermal      (a)             ND               (a)

Pretreatment   Lignin     Changes
               removal   to lignin
                         structure

Diluted acid     --         (a)
Alkali           (a)        (a)
Hydrothermal     --         (b)

(a) Higher effect. (b) Lower effect. ND: Not determined.
Source: Adapted from Mosier et al. (2005).

Table 2. Physicochemical characterization of grinded
and dehydrated orange waste.

Crude fiber (%)          Moisture (%)            Ash (%)

12.23 [+ or -] 2.95   10.05 [+ or -] 0.10   3.46 [+ or -] 0.07
11.04                        12.16                 4.92
7.17                         0.96                   --
--                           13.36                 8.57

Crude fiber (%)          Protein (%)           Lipids (%)

12.23 [+ or -] 2.95   3.38 [+ or -] 0.69   1.74 [+ or -] 0.28
11.04                        4.85                 2.16
7.17                        11.08                 6.00
--                           7.93                 3.44

Crude fiber (%)            Total             Pectin (%)
                      carbohydrate (%)

12.23 [+ or -] 2.95        69.14         12.03 [+ or -] 0.55
11.04                      70.08                 --
7.17                         --                  --
--                         68.85                 --

Crude fiber (%)             Reference

12.23 [+ or -] 2.95       Current assay
11.04                 Ruviaro et al. (2008)
7.17                  Clemente et al. (2012)
--                     Retore et al. (2010)

Table 3. Experimental design of the acidic pretreatment
with orange residue.

Assay    Time    [C.sub.acid]   [C.sub.biomass]
        (min.)       (%)              (%)

1         15         1.0              1.0
2        120         1.0              1.0
3         15         1.0              9.0
4        120         1.0              9.0
5         15         5.0              1.0
6        120         5.0              1.0
7         15         5.0              9.0
8        120         5.0              9.0
9        67.5        3.0              5.0
10       67.5        3.0              5.0
11       67.5        3.0              5.0

Assay          TRS *                  MY **
                (%)                    (%)

1       52.29 [+ or -] 2.03    41.43 [+ or -] 4.82
2       65.52 [+ or -] 14.50   36.02 [+ or -] 4.34
3       26.37 [+ or -] 1.08    41.15 [+ or -] 1.48
4       39.59 [+ or -] 5.82    40.14 [+ or -] 5.06
5       62.53 [+ or -] 11.63   41.16 [+ or -] 3.13
6       56.70 [+ or -] 2.79    32.90 [+ or -] 3.61
7       32.89 [+ or -] 6.66    39.60 [+ or -] 0.07
8       33.95 [+ or -] 9.24    34.73 [+ or -] 1.29
9       26.40 [+ or -] 5.79    34.83 [+ or -] 0.15
10      28.68 [+ or -] 1.45    35.82 [+ or -] 0.19
11      29.30 [+ or -] 0.57    30.62 [+ or -] 6.22

Assay        TSS ***
               (%)

1       1.65 [+ or -] 0.07
2       2.23 [+ or -] 0.05
3       0.59 [+ or -] 0.02
4       0.70 [+ or -] 0.00
5       6.62 [+ or -] 0.11
6       7.06 [+ or -] 0.19
7       0.97 [+ or -] 0.02
8       2.11 [+ or -] 0.01
9       1.73 [+ or -] 0.02
10      1.72 [+ or -] 0.06
11      1.71 [+ or -] 0.07

Results are expressed as mean standard followed by standard
deviation. * TRS--Total reducing sugars; ** MY--Mass yield;
*** TSS--Total soluble solids. The three studied variables
influenced sugar and total solid solubilizations, though
their effects were not significant in the studied intervals
for mass yield. Positive values in the Pareto charts indicate
positive contributions of increased variable within the studied
interval, whereas negative values indicate negative contributions.

Table 4. Experimental design from alkaline pretreatment of
orange residue.

Assay    Time    [C.sub.alkali]   [C.sub.biomass]
        (min.)        (%)               (%)

1         15          0.5               1.0
2        120          0.5               1.0
3         15          0.5               9.0
4        120          0.5               9.0
5         15          2.5               1.0
6        120          2.5               1.0
7         15          2.5               9.0
8        120          2.5               9.0
9        67.5         1.5               5.0
10       67.5         1.5               5.0
11       67.5         1.5               5.0

Assay          TRS *          MY **        TSS ***
                (%)            (%)           (%)

1       17.07 [+ or -] 6.34   31.14   1.80 [+ or -] 0.07
2       23.08 [+ or -] 6.59   25.81   1.70 [+ or -] 0.05
3       1.27 [+ or -] 0.34    81.30   1.00 [+ or -] 0.02
4       1.51 [+ or -] 0.22    69.91   0.10 [+ or -] 0.00
5       12.00 [+ or -] 5.44   28.09   2.80 [+ or -] 0.11
6       19.26 [+ or -] 7.26   29.42   2.30 [+ or -] 0.19
7       8.36 [+ or -] 1.40    27.24   6.60 [+ or -] 0.02
8       14.88 [+ or -] 2.79   27.67   5.80 [+ or -] 0.01
9       14.31 [+ or -] 6.83   30.30   2.80 [+ or -] 0.02
10      14.07 [+ or -] 4.82   29.00   2.80 [+ or -] 0.06
11      11.29 [+ or -] 4.70   30.78   3.40 [+ or -] 0.07

Results are expressed as mean standard followed by standard
deviation. * TRS--Total reducing sugars; ** MY--Mass yield;
*** TSS--Total soluble solids. The three studied variables
influenced sugar and total solid solubilizations, though
their effects were not significant in the studied intervals
for mass yield. Positive rates in the Pareto charts indicate
positive contributions of increased variable within the
studied interval, whereas negative rates indicate negative
contributions.

Table 5. Conditions and results obtained in the hydrothermal
pretreatment of orange residue.

Assay    Time    [C.sub.biomass]          TRS *          MY **
        (min.)         (%)                 (%)            (%)

1         15           1.0         37.35 [+ or -] 5.49   51.50
2         15           5.0         6.41 [+ or -] 0.50    56.00
3         15           9.0         3.43 [+ or -] 0.08    59.46
4        67.5          9.0         25.05 [+ or -] 0.84   47.92
5        67.5          1.0         5.75 [+ or -] 0.27    52.87
6        67.5          5.0         3.08 [+ or -] 0.13    55.55
7        120           1.0         37.02 [+ or -] 4.65   46.11
8        120           5.0         8.92 [+ or -] 0.81    52.37
9        120           9.0         10.97 [+ or -] 1.50   55.76

Assay        TSS ***
               (%)

1       1.80 [+ or -] 0.07
2       1.70 [+ or -] 0.05
3       1.00 [+ or -] 0.02
4       0.10 [+ or -] 0.00
5       2.80 [+ or -] 0.11
6       2.30 [+ or -] 0.19
7       6.60 [+ or -] 0.02
8       5.80 [+ or -] 0.01
9       2.80 [+ or -] 0.02

Results are expressed as mean standard followed
by standard deviation. * TRS--Total reducing sugars;
** MY--Mass yield; *** TSS--Total soluble solids.

Table 6. Summary of the significant factors for each
pretreatment.

Pretreatment              Variable                Time

Acidic                      %TRS                positive
                           %MY *                   NS
               [[degrees]Brix/g.sub.biomass]    positive
Alkaline                    %TRS                positive
                           %MY *                negative
               [[degrees]Brix/g.sub.biomass]       NS
Hydrothermal                %TRS                   NS
                           %MY *                negative
               [[degrees]Brix/g.sub.biomass]       NS

Pretreatment              Variable              [C.sub.acid/alkali]

Acidic                      %TRS                        NS
                           %MY *                        NS
               [[degrees]Brix/g.sub.biomass]         positive
Alkaline                    %TRS                        NS
                           %MY *                     negative
               [[degrees]Brix/g.sub.biomass]         positive
Hydrothermal                %TRS                        --
                           %MY *                        --
               [[degrees]Brix/g.sub.biomass]            --

Pretreatment              Variable              [C.sub.biomass]

Acidic                      %TRS                   negative
                           %MY *                      NS
               [[degrees]Brix/g.sub.biomass]       negative
Alkaline                    %TRS                   negative
                           %MY *                   positive
               [[degrees]Brix/g.sub.biomass]       positive
Hydrothermal                %TRS                   negative
                           %MY *                   positive
               [[degrees]Brix/g.sub.biomass]       negative

* MY- mass yield (%); NS--not significant.

Figure 1. Sugar saccharification, mass yield,
and solubilization after acidic pretreatment.

Standardized Effect Estimate (Absolute Value)

TRS (%)

(3)[C.sub.biomass](%)(L)   -19.5
Time (min)(Q)              14.21
1(L) by 2(L)               -5.87
(1)Time (min)(L)            4.07
1(L) by 3(L)               11.29
(2)[C.sub.acid](%)(L)       0.43
2(L) by 3(L)               -0.10

TSS

(3)[C.sub.biomass](%)(L)   -19.4
(2)[C.sub.acid](%)(L)       17.0
2(L) by 3(L)               -11.8
Time (min) (Q)              6.26
(1) Time (min)(L)           3.34
1(L) by 2(L)                1.30
1(L) by 3(L)                0.35

MY (%)

(1) Time (min)(L)          -8.05
Time (min)(Q)               3.02
(2)[C.sub.acid](%)(L)      -1.62
1(L) by 3(L)                1.21
l(L) by 2(L)               -1.05
(3)[C.sub.biomass](%)(L)    0.54
2(L) by 3(L)               -0.56

Note: Table made from bar graph.

Figure 2. Sugar saccharification, mass yield,
and solubilization after alkaline pretreatment.

Standardized Effect Estimate (Absolute Value)

TRS (%)

(3)[C.sub.biomass](%)(L)    9.37
2 (L) by 3(L)               6.05
(1) Time (min)(L)           4.13
(2)Calkall (%)(L)           2.34
1(L) by 2(L)                1.55
1(L) by 3(L)               -1.34
Time (min) (Q)             -0.90

TSS

(2)[C.sub.alkali](%)(L)     15.2
2(L) by 3(L)               11.40
(3)[C.sub.biomass](%)(L)    5.76
(1) Time (min)(L)          -2.70
1(L) by 3(L)               -1.26
Time (min)(Q)               1.17
1(L) by 2(L)               -0.35

MY (%)

2(L) by 3(L)               -26.4
W[C.sub.alkali](%)(L)      -26.2
(3)[C.sub.biomass](%)(L)    25.0
Time (min) (Q)             11.46
1(L) by 2(L)                5.05
(1) Time (min)(L)          -4.08
1(L) by 3(L)               -1.90

Note: Table made from bar graph.

Figure 3. Sugar saccharification, mass yield, and
solubilization after hydrothermal pretreatment.

Standardized Effect Estimate (Absolute Value)

TRS (%)

(2)[C.sub.biomass](%)(L)   -9.28
[C.sub.biomass] (%)(Q)      4.89
Time (min)(C)               2.38
(1) Time (min)(L)           1.10
1(L) by 2(L)                1.09

TSS

(2)[C.sub.biomass](%)(L)   -11.6
[C.sub.biomass](%)(Q)       2.76
Time (min)(Q)              -0.61
(1) Time (min)(L)          -0.60
1(L) by 2(L)               -0.08

MY (%)

(2)[C.sub.biomass](%)(L)    7.99
(1) Time (min)(L)          -4.37
Time (min)(Q)               2.48
[C.sub.biomass](%)(Q)      -1.62
1(L) by 2(L)                0.71

Note: Table made from bar graph.
COPYRIGHT 2015 Universidade Estadual de Maringa
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2015 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Title Annotation:texto en ingles
Author:de Farias Silva, Carlos Eduardo; Silva Belo Gois, Georgia Nayane; Oliveira da Silva, Livia Manuela;
Publication:Acta Scientiarum. Technology (UEM)
Date:Oct 1, 2015
Words:6634
Previous Article:Parameterization effects in nonlinear models to describe growth curves/Efeito da parametrizacao em modelos nao lineares na descricao de curvas de...
Next Article:Reaction of dissolved ozone in hydrogen peroxide produced during ozonization of an alkaline medium in a bubble column/Avaliacao do efeito da reacao...
Topics:

Terms of use | Privacy policy | Copyright © 2019 Farlex, Inc. | Feedback | For webmasters