Boresurce Tecnology 199 (2016) 173-100 Contents lists available et Scieneabirot ca io Bioresource Technology journal homepage: www.elsevier.com/locate/biortech Steam explosion pretreatment of oi] palm empty fruit bunches (EFB) using autocatalytic hydrolysis: A biorefinery approach Jesus David Coral Medina*, Adenise Woiciechowski*, Arion Zandona Filho®, Poonam Singh Nigam’, Luiz Pereira Ramos‘, Carlos Ricardo Soccol Shoot of Boreal Sciences. University of Uber, Coline. UK Fodor Unicity of Pt Department of Chest, C815 97D Carib, PRR HIGHLIGHTS s Anaysis of peetcstment step using steam explosion under autacatalyticconetions {THrosin in CUEHS sructures after seam explasian, myprowog the povesty aod tecucing the cystalnily of cellulose { Solubilsaton of hemiceluise in form of xylose and arabinose 2‘ Eovilunens of 24% in cellulose per gram of OPEPS este. {Production of hire! hydrolysate en fermensabiesgars, ARTICLE INFO ABSTRACT ‘The ol palm emp fn bunches (FF) are an arcactve snr af avon fr the prncieson of biochem seal products, therefor, the ssh ofthis Wore to analyze tne eet ofthe stent explosion (SE) pretest ent under dutecatlvic conditions on EF usin 2 full experimental design. Fem me were the operational varables studied, The FFB treated at 195°C lor 6 in showed an inezease of 34507 in glycan ‘mostly cellsose) and 9 reduction oF 612% In emicelloloses, wth incrmased enaymatied gestibity to 33% producing 42. Lt gluease, Seanining een meragrap of thesteary ested HHH exhisited suriaee eros ane an inereaeee ser porns, Faurer transform anfared spectrascupy showed the solubilization ot hemiceIslose and medication of celluose in reared EB. 1 2015 Hlsevier tr All ight reserved Avsiabe enine# Sepentber 2015 Co gai ery ut bunches 1. Introduction produced generates 1.1 tons of EF (Shinoj et al. 2011}. This lignoceliulosic biomass is mainly composed of cellulose (glucan) emicelisis the production of oil palm from Eets guincensis in Brazil has increased in last decade, being the 10:h preducer in the world. However, $0 of 0! palin in Brazil is used inthe food industry. With the launching is anvary 2010-01 the Brazilian National Program for the Production and Use of Biodiesel (PNP), the inelusion of oi palm 4$ Ieedstock for biodiesel production 31.8 cullion hectares (ha) of forested area were identified in the Northeast, wich good soi) and climate concitions for palm off cultivation. Mozeover, che local installation aad operation of almast 13 plot plants improved the Production capacity to 751 million of titers of oil per year ‘ogether with oil palin production, the generation af selid waste calic¢ oil palms empty Trait bunches, commonly referred as (EPR) is increasing. ts estimated that each 1 ton of oll palm Fat aber: socclBulprcorn be (CR. Socal lb ong/101016) bortet:2015.08, 125, {xylan and arabinan mainly) and lignin. Seing an attractive source for biofuels and value acdeé chemicals it ean be Included inte energy matt te improve the sustainability of palm oil biorefineries (Gnansounou and Dauriat, 2010; Noomtim and Cheirslp, 2011) ‘ecatse ofthe recaleitrant nature ofthe lignocellulosic material foune in FFB, a pretreatment stage is requited t0 solubilize the hemicellulose and lignin and enhance the mass proportion of cellulose in the biomass Pretreated material will have ao increased porosity and impravee accessibility toward the subsequent Liydrulysis process, forthe production of sugars useful for ethanol fermentacion (Saikku et al, 2012). Steam explosion (SE) has been proved to be effective for a vat ly af lignocellulosic biomass, including haréwoods, saftwoads, herbaceots resictes, sugarcane bagasse, and wheat straw, being the most widely employed physico-chemical pretreatment or1 106. Media 2 Rereante te any Fgnocellulosic biomass (Fernandes et al, 2015; Ramos et a 1992}, During SE, the biom: with saturated ecompressinn. The high pressured steam provides an adequate vehicle to heat biomass, penetrating ane! modilying the cell wall structure, ensuring the effective hyécoiysis of mainly hemice! jases making cellulase more accessible. The sueden decompression causes an adiabatic expansion that makes the material t0 8 mechanical diseuption of fibers modifying the plane cell w. (Hendriks and Zeeman, 2009; Mosier et al, 2005; Ramos, 2003} inthis study, she effect cf steara explosion (SE) on the fraction. ation of EPH was evaluated through the application of a full experimental design (7), using temperature and reaccion rime as so main operations\-vatiables, Changes in she mass composition ff cellulose, hemicellalases and acd msoluble hgnin in. solid faction after SE were charscterized anc the liquid fraction was analyzed! far the concentration of ferstentable monamerie sugars Ane the generation af organic acids andlor inhibizory compounds, The effect of SE on EF riber was measuted by the digestibility percentage, while the chemical modification of BEB was analyzed by Fourier Transform infrared spectroscopy (FIR) and the structure madifeation of FFB fibers was studied by scamming, election microscopy (SER!) 15 heated for a shoct time period 2. Methods 2.1, Raw material 0 palm empty fruit bunches (OPEFB) were obtained from Bio- palma Vale factory, located in Moya, Para, state of Brazil, The OPEFB wsas eriee ima cross Taw stave ar 65 for 72 hand stored in poly urethane bags at room temperate te avoid biological degradation. 2.2, Characterization of the EFB pretreatment fractions The composition ofthe raw EF and the soli Iractions obtained! after SF pretreatment, was determined accosding to the NRFL ana iytical procedures reported by Sluiter et al, (2071), he cellulose {Cet} and hemicellulose (Hem) mass composition were calculated using the equations reposted by Tan et al. (2013), in which the concentration of glucose (Gl), xylose (Xyl} and arabinose (Ara) are correlated, Analysis of acié yralysate was carried aut by ligh-Petiormanee Liguid Chromatography (HPLO), in @ Shimadct, Chromacograph equipped with an Aminex HPX-A7H colar swosking at 60°C with sulfuric acid (5 mmo! ') as mobile phase at flow tate of Grab min | Furfural (F), hydroxymethyl furfural (HM) acetic, formic and levulinie ace were determined in the pretresiment hyerolyzates ising a Shimadze Chromatograph equigned with an Aminex HPX-871 and C18 columns at 69°C. Mobile phase used was sulfu- ie acié (5 mmol L-') at race of G mil min” with an IR detector Decection was carried aur by eiffrenris) retractomesny and oho ‘quanbilieation vas based on external calibration as described by: Scholl et al. (2015a). The acid soluble lignin (ASI) andl acid insoluble lignin (ADL) were éotecmined as suggested in the NREL procedure (Sluiter et al, 2011), The acié soluble lignin was measured by UV spectroscopy a1 280 nm using a SP - 2000 UV speccrophotuneter with dilation factor of 10. vihile the acil insoluble lignin corresponded to the ash free residue that was obtained after sulfuric acid hydrolysis 6% plans polysaccharides 23, Ash determination The ash eeterminstion was carried aut following che procedure seposted by Sluiter et al. (2008) wih litle modifications. Bre! 1.3 g of dry 88 were placed inro a enucible and the samples were calcined at 555°C fer 6 24, Protein determination The quamitative devermination of soluble proteins was performed using the protocol repoctad by Hames etal, (2008). 25, Exteaetives methanol and water The determination of extractives components was cattied out according, to the NREL procedure seported by Sluiter et al. (2005). All experiments were done in uiplica 26. Steam explosion pretreatment (SE) The SE presreatment was carticd out in 2 stainless stec! zeactor with a 101 capacity. The process Row diagram is presented at Fig. 1. Pretreatment was performed with 300 g of almast dried FYB, containing 202.0.7% moisture. 1t has been demonsieated that bumidity has no inflvence an the recovery of said an Hique on obtained afler SE (Pitarelo et al, 2012) cial was intteduced in the veacter vessel and saturated m was fed unt the desired temperaiure was reached, the heating Lime in all cases was around 2a. The reaction time was controlled etter the temperature was reacieed. The sudden decompression releasec the material into a cyclone and the vapor was Hberated to the atmosphere. The pretreated maternal was washed five times with water (11) and salids were recovered by {centifugation. The fist vo washings wese collected for analysis The solids wore oven dried and + faction was millee for eatboiy- rate and lignin analyses. 22. Screening of preteeatment conditions tke influence of temperature ané sime on EPL pretreatment was evaluated initially ab 160°C andl 252°C for a Fxed reaecion time of min. The PFW bets denved trom pretreatment were chatacterized in tess of cellulose, hemicelllose and lignin. The rrorpholagical changes were followed by SEM, with the aim to check the influence of the severity of different pretreatments ton FRR, 28. Experimental design and statistical analysis From the screening stady, @ full factorial design was imple rented, with three level and two operational variables (J) Temperature and teaction time, with tree replicates af he center point to evaluate the systema ervar. The recovery of solids and their mass composivion in terms of cellulase, hemicellulose and acid Insoluble ligran were expressed 35 response variables, The result ing values and stacstical analysis were processed using Statistica Version 7.0 (Minneapolis, USA). Analysis of variance (ANOVA) was employed co determine statistical significance of the macel fhe experimental response obtaived was analyzec with a second-order pelynomia! as 1s presented in Fa, (1) The surmmary of experimental design is presented in Table 2 KTS ST BAX 1 where y is the response (cellulase, hemicellulose and acid) ‘insoluble lignin in fiber‘): X' apd Xj are the independent variables ar fj Bi hy ae She intercept, linear, quaerati and interaction coellicients, respectively.er vied — wou sem a wean * Heat bet g Exchanger ele >t ite mm > > reas weer au ia. 1. ew azn of ea eaplacion press done om he severity ator index or each conion of temperature and tie was omhuted uit equation ported by Oweoad wea (1987) which presence in Ea 1:00 Cars) che aceon ofeach wos sgl stan "uses of these pretseatine % [ exp 2 alyzed in terms of glucose, ‘urfural, yéroxymestyl furfural, acetic, formic contents with 2 hydroiyzaves in biorefnery conceps Hythe possible 2.8, Enzymatic hydrolysis of FFB treated The eiciency of steam explosion igestbilty of FFB treated sn enzymatic hytralyss. The sachari cation was developed in accurdznce with the LAP of NREL ‘Selig ct al, 2008} The untreated ans. treated SFB at temperature of 5, 185 ang 195°C maintaining for Gmin were used. The commercial enzymes usec were Celluclast 11, produced by Trichoderma reeses and Novozymes 188 prochiced by Aspergills niger, The enzyme sample used for activity measurements vaas measured by ie contained a 1:0.30 mass ratio of Celluclase 1.5L. ane) Navozymes: 188, respectively as reposted by Silveira eta. (2014). The activity bf the enzyme losded was 10 FPL per of ceiitlose adc low Ing the procedure ceported by Ghose (1987). Th wos analyzed for the quantity of ghicose released in 120 reaction ay éinitro salicylic acié (DNS) assay. The digestibil wes determines using the equation repored by Selig. et (2008), All experimenis were earried ur using varee reaticate mre 2.10, Fourier transformed infrared (FTIR) Spectroscopy The FTIR analyses of the samples were eared out using a varian equipment with a vesolution of 4 cm | and 32 scans per ‘minute, The biomass pellets were obisined by mixing snd pressing, ir with KSr at 0.1% of biomass. The range of wave memes was se" from 400m "x 4000 cm 2.4L, Scanning etectron microscopy (SEM) The mor BB was monitored! by SEM, ander SCAN VEGA 3 MU, with, ology of treated § nitrogen avnosphere in a microscopy18 1 an acceleration voliage of 15 kV. The resolution of 3 nm, anc che image magnification was 2500X, The samples were vacusim dried alized with gold ina sputter cnarer and dred evernight before analysis 5. Results and discussion 3.1, Biomass composition The biomass composition of EER an dry basis before steam explosion pretieatiaent was as following. etlanol extractives 479+ 1.33% water extractives 5.97 = 55%, lipids 625 +1112, proteins 336+ 1.23%, cellulose 78,00+907%, hemicelulose 24.12 «0.07%, acid insoluble lignin (AIL) 17.84 © 1.64%, zcid soluble ignin (ASL) 2.12 =0.0%, acety! group 520 0.98% ash 3.19 = 0.07% hased en the characterization done, correlating the glucose, aylose and arabinose as cellulose and hemicellslase, the total polysaecharide content present in EB was 52.12 vt, being git cose and xylose the main consticuents of cellulose and hernlceliue las, respectively, The fora lignin action was 19.98 we wath acid insoluble lignin being its main component. «conditions 32 Screening of pretreat The influence of SE on FFB was analyzed al two distant severity factors (102 Ky). [he results have Neen summarized in Table | The cecovery of solids decreased drastically with increasing pretest: ‘ment severity, However, the ma ge of ceitulose in the pretreated EFB fibers increased by at least 12% at 212-C. The nyeleolysis of hemicellulose was ditectly. proportional toh ‘peranal conaions Tine ann Teoperae etna ermowesteenolagy 195 (2016) 17s 180 Increase in pretreatment temperature. Ata severity of 42, 86% of he oiiginal hemicellulose vias solubilized, genetating ETB fiber residues with mass compositinn af only 3.93% of heanicelhloses, results obtained ia this work {yom the sereeniag activity were compared with those reported by Baharuddin et al. (2013) for steam pretreatment at 230°C and 10 myn, the percentage of mass Joss was 3.8% lower, witeress te increase in cellulose mass com position was 6% luwer. Taese results st be considered important for a biovefinery pretreatment step, decause fibers that are sich in cellulose ane jaw in hemicellulose content are attractive for their subsequent biotransformation, in value acidee chemicals such as ethanol, butanol end organic aces, On the basis 0° changes in the mass composition and the fiber siorphology of the EFB alter SE (See Supplementary data $1),a full factorial design was bull wath the aim of finding the ideal conc: ions to pradice FFB fibers rich in cellulase, low in hemicellulose, with a higher cell wall porosity and with a noticeable reductions sn their eysalinty 5.3, Steam explosion experiments The full experimental design and heir cespective global mass balance in both solid ane quid pretreatment fractions are pre- sented at Table 2, The solid vecavery decreased with inevea naperatute and lime, being ateater the efTec of tem eratui that of time, walle che mass ir hyttralyzate increased with tersper- ature and time. These results are importane flem the industual ‘view point, beeause best treatment condition must provide 2 goed relationship petween the solié recavery and the cellulose concent available in the pretieated biomass “The mass composition of EES after each run of SE was carried bout in terms of cellulose, nemicellulose, acié soluble lignin (ASL) and insoluble lign's (AIL) as described in Scetion 2.3, the resules are summarized in Table 3. Tae mass composition of cellulase in tit peaked at 34.89 wee at 195 °C for 6 min al higher severities Jed to substrate lett with lower cellulose content. This is cue 10 1e progressive hydrolysis of plant golysaccharide to monomeric sugars and 19 the subsequect production ef degradation com pounds such as furfural, hydroxymethyl furfural, levulinic, acetic and fore acid Tre mass composition of hemicellulose in FFB decreased gradually with an inccease in presreatment severity, which was a aaa desiree lige: in this pretreatment scudy. By contrast, the mass fed sue grin poo 2004 composition of AI. in steam treated suhstrates increased wis ea asolbie gra asst pretreatment severity, mostly due 1a bemicellulase removal by * Composiion em ary has acid hydrolysis. However the accarrenee of condensation reactions " " Snvolving lignin and ether materials, suet as tannins, furans and xpesewtal design an lob mas balan" nec exgesiesta cule Gave Revue” voce vake assis) mass ig) sttraey® —rystatep) recovery (8) 4 soo 1 cet his 439 5 wo 8 ° xa ae n33 oo 18 a fea * 1 ° a a3 brit 0 ° 0 aa ait "2800. Westin ete tersoureTecmnolegy 196 2016) 175-180 a Ron Callow) Hemicelwine (2) ASL) AL ake 425 03 a0 pherotie compounds cannot be ruled aut as already reported by fther authors (Haghighi Mood et al, 2013; Sun et al, 2012) I genera, SE caused an increase ta 0.25 g of glucans per gram fof FFB, which represents 24,6% more celiilase in relation wo the untreated EIB. The hemicellulose quantity was reduced 10 0.08 g per geam of BFB aud this corresponds to 68.115 less hemicellulose compared 0 found in untreated FFB, These resulis are more promising than those reported by others using steam explosion for FFE pretreatment (Baharucidin et al, 2013; Shamsudin et al 2012), ‘0 confirm che effect of each operational variable om each response variable, analysis of variance ANOVA was developed with significance level of p< 005, The summary of the effects of ‘operational variables an each response, the pure eeror and the conelatioa Coelficient 4?) ere summarized i Table 4. the rmathersatical model adequately represented! the solid recovery, cellulose sad hemical ulose mass percentage in FFA after SE, The lack of fi in acid insolubie lignin. would be explained by the increase in theit mass composition as discussed above toe Tiveat elect of both temperaiuse and time had a sisung influence on the Fesaense of all variables, The quadratic eect of both: semperature ard time was more remarkable for sali recow- ‘cy and hemicellulose mass composition, These effects and sig ieance levels were important to generate 2 mathematical mode that was able to descrlve the SE effect of BFR, The mathematic rode! 20 describe the effects ef SE process or cach response variable are presented in Eqs, (3)-(5) as function of the pretreat temperature (1) and seaction me (6, where $8, C,H, All, are the reeavery of preiteatment solids and the cellulose, hemicellulose and acid insoluble lignin mass percentage ff steam eated EPB. The temperaiuse (7) snd ine (¢) are given Jn Celsius degree ane minutes, respectively. These equa Important for modeling and simulation of SE, in order to perform scaling pracess ort» develup 2 sersibilty analysis af the applied Diorefinery conceat. “The response suifaces of these models are presented in Fig. 2 Fig, 2a shows the elfect of increasing the reaction time and temper aluse on the cecovesy of solids pretrcated, which decseased wih the inerease of temperature and! reaction time, Fig, 2h, shows the response surlace for he substrate cellulose content. This Ligu shows an optimal point, whieh comesponds to the great Increase in eellsiase content after SE at 198°C tor 6 min, Further Increases in pretreatment temperature procuced 3 decrease in the substrate eellolose content, mainly de to hydrolysis degradation to products as furans, evulinic, formic and acetic acid (Larsson et al, 1999), Fig, 2c, shows the response surface for the rate hemieaalose content in relator: co changes i tempera ‘ure and seaction ime, The increase of both time aad temperature decreased the mass pezcent of hemicellulose ia BEB progressively, indicating that more severe pretreatment coneitions caused a con. siderable mass loss of plant polysaccharides. Funaly, the increase: in the ace) insolusle Iigein (AIL) of FFB Abers is presented in Fig. 2d. This phenomenen is induced by hyctrelysis and condensa tion reactions as discussed above. Therefore, the authors suggest a delignification process after steam explosion to improve the performance of EFS in a biotefinery factory. 3.4. Analysis of the pretreatment figu hyatrotyzate ‘The frst two liquid fractions derived trom water-washing of EAB were characterized by HPLC. fa general, monomeric sugars were detected only in the first liquid fraction, while the 20% of tHe total concentration organic acids as levine, forse and ace acid, furfural, hydroxymetayl forfacal (HIME) were recawered in f= trate of the second! washing stage. The summary of this qualitative analysis 1s presented in Table 5, The results are aligned with the progressive mass loss that was abserved at higher pre severities. These losses weve primarily associated with hemcellu lose hydrolysis since the concentration of pentoses such as xylose ane arabinose ia washings inevensed tageches with an inerease in Temperature aad ceaction time. These results are in agieement SR) = 301.03 1.69. T+2.70- 107 7166 1-005 with the pretreatment mass balance of sugarcane bagasse as (3) presented by Rocha eal (2012) and Scholl eta (20130) Re concentration of acerca liquid yaolpate neease © =-24120042545.7.-007.78--1865+4 ty the seventy te mainly eee of atoetalye hyarotyts tie {oyetier wth the deacetylation of henscelllose and be he all ini 4 highest concentration of 5,241 ' of acetic acid were detected at . . 195°C for nin These concetation represent the 61.9 of ae H(%) = 90.87 014-7: 36-1077 9.38.6 ‘group present in untreated EFB, The values obtained in this work 058. 69) 10 Tt {5) are simiae thowe reported by jg ets (2013) i ote sed Dreeatneat of fi) 23597 -227-1 90-10% 7 173-4436." Sonenin of mena ony compen Poet 6 ty of pretreatment, the highest concentration were Tate Soe “so ae 2 a 10> i tar vert a am 2 33 130 an 3a Fin 2. Sse tess of a Saas ane tiates IN ses of tse fom a Fete afer on expen. 5) Sel ecoveny espe (Cais rsponse (Het (ellos espns 0) Ad sie i mess aun laces gf) Stove (gt) _Aabinose (a) Acetic ad (gi) Fanlal )_AVEF (UM Feri ace Gi) fev lh na. Not detected git and 0.23 git 195°C for 8 and 10 min, espeetively Conventratians are lower chan that reported im iterate 3s tory for ethanol fermentation by Saccharomyces cerevisioe 2 (Almeida et al, 2007; Palmqvist and Hahn-Hagerdal, 2000). The presence of levulinic ang farmvic acid in liquid hyérelyzate increased with the severity factor of pretieatment, indicating the lhyeravion of the furfural ane HIM, with the increase in che sever ff treatment, The concentration of formic acid, was alo. Sant forall conditions of pretreatment, tne highest conc 01 O21 gL. On the other hand, the highest concentration of OTL * of levulinic acid was detected et treatment of 195°C for 10min. These concentrations are lower than the vale commonly reported in biomass iyéralysis (Almeida et al. 2007) 25, Fnzymariccigestibility of FFB 1rsated by steam explasion The percentage of digestibility was determined in g of cellulose EFI and SE-Wealed B°B at the runs 1, 4,7 (Table 2), ‘he results are summarized in Table 6. These results revealed that the digestibility was improved with the merease in the severity of pretreatment, 195°C for Grain being the best condition with a Aigestility of 124 oF FFB. The es hye of Bat: on, aso produced a syrup with 4.186 fof glucose, these results cepresent simest twice the glucose proclaced tram enzymatic Hydrolysis of untreated and seated EPR at 175 and 185 “C. The wesults obtained in this work are higher than reporiee by Shamsudin ef al, (2012) using lowest enzyme concentratian and reaction time, The percentage of digestbilsy of untreated EB was slshtly higher thot the tteatce FFB at 175 ane 185°C, These results could be explained by the presence of depclymerized low molecular \veight lignin, tannins, saponifieanle extractives, pat of hemsicell: Joses and otter degradavion predivets trem FR her sur:ace arsen uring the steam explosion preteatmenc cat can overcome the elect of availability of collslose in treated FFB, with respect to luniteated BFS (Fernandes et al, 2015)Sui SSoincktia S,Pngs 2017 Ang anos change cat Tnoyisotsdo-eiot poratesGrnramamowa siairden St 8 Scisbanlitiene) , Mea’, RRaGE Ro, AZ, Rane ROR A unease Tyger aay Tremor roma PURER on CH ce clip gs fates sine ele ia pee Shalt MeegelD. Prarie APRAAGARC. fl. AZ. amos hi. A EGR hg aaa eC PAN pretest ny seam sxpiasa. “Technol 82, 226-237 hp oiorlta 10g! urea 201508 068 xi. eect AB) 1510, itt CSR ARE Sa RISE RE ASE ie iSitnadereiesormccuttlopepplemenrsty daca ©2)r his, ‘ete ere Fe ei! en em STE Sy RRS atcea nae WE Sithindy act 73"C, PSC sie Nan ach eran. Kis the, sprain Seraterncati dure ot ieee tier preseitets oe. With the increase in St temperature the bands a 2869 eon and'2799 em "ate reduced. These bands correspond to the C~H Stretching vibrations of aliphatic moieties mainly in plan polysac les such asce!ilose and hemeelafos, A stretching vibration 50569 em vas present nal samples ane this was ateited %9 Cin tighin strctures a aleady reported by Nieves et 1.2011) The band at 1461 em? and 1415 cm are more emariable with te increase of severly of pretreament, these could bed 1 cel- lilose as proposed by Wan et al, (20001, The band at 1530 cm represents she non etherfed OM in ignn sectes specially in Syingy units, his band i modified with the emperature increase, Confsming the mocifcation in lignin srvture. as ceporced by others (Laurchesse-andAvérous, 2013 Mansour and Salad, 3006). Te vibration at [250em | disapeared with te tempera cane of SE twas related with ring breton a €-OF se sts and glycosidic bonds vibration typical of aylans. The band T1G3cm represent ihe C-O-C stretching th amozphous cellos with the mdifcaion ef cellulose By SE. Finally the band at Toabcm-" deceased with the seventy of pretreatment. i cole be ascribed 1a pelycsiele lskages a2 was mnetioned by Als etal (2009) 4, Conclusions The EPB was teated by autecatal best pretrestment perlormance was achieved at Grin, with increase of 24% ia cellulose and 68% ¥ newiceltulose. The wea an enaymatie digestibility of 33% that produced! a hydrolyrate with 42g" of glucose. The hydrolysates consisted of glucos xylose ane arabinose as lermentable sugars, with high acetic Acié contents. This suggested che possinility of 3 subsequent Sseparationjtransformation pracess to oblais precursors for value-added products, enhancing the sustainability of BFE biovefinesies. fac steatn explosion. The 195°C for ction in Acknowledgements ‘The authors would fixe to extend their sincere gratinide £0 VATE ane VALE International Inc. for their financisl support sad for sup- diving the EFH simples, This work was alsa supported by the grant ‘rom the Organization of American States (OAS) ane CAPES ro the fist author, We also acknowledge Conter of the Federal University af Parand for ch the morphological analysis of FFB. pp resets use gh 99 209 2/5 28180 a ‘AipenaBloremeppletivedtalfmateri | cice/0 30165 Anne te aegp etnias! sluer A Ruz Sata. Shiter, J 2008, Deerinatiop of Exuactes ig ioSiod Biatady wacom sttons an ska Be kn sone version, a hip: /dxlo.org! 10.1016 boetech.2015,08, stg He, Bo le Keck CS. ean. 200 Srinaton of Ash n Biomass taboratnry Anaya! recede (UN) Golden, Color. stator Aromes 9. Ruz R. Scat. Ser Templeton D. Crocker, 20 Hefacanaim of Stara! 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