Toumal of Food Technology 3 (1): $35 537, 2008 ‘© Grace Publications, 2005 Ad ive Effect of Suybean Curd Residue, Okara, for Enhancement of Methane Production from Pretreated Woody Waste “Harumi Take, ‘Godliving Muui, ‘Fu ‘Graduate School of Nat Kanazawa University, Kakwna-inac sist Kobayashi and 'Yoshitoshi Nakamura al Science and Technology, i, Kanazawa 920-1192, Tapan “Applied Microbiology Unit, University of Dar es Salaam, P.O, Box 35060, Dar es Salaam, Tanzania Abstract: In orr fo convert woorly waste info methane gas efficiently, the pretreatment elect of steam ‘explosion and the additive effect of aoy'bean curd ro due, okara, were clarified. 1¥0 mL of methane gas was ‘obtained from 1 g of steam-exploded Japanese cedar chips ata steam pressure of 4.51 MPa and a steaming time of S min while no methane gas was produced fr attempted ancl the optimal eawdition n untreated chips, The addition of okara into dhe chips was CIN of 18, increased the methane gas peesbicerl ty 15 ml ‘Key words: CIN ratio, methane production, woody waste, soybean curd residue, steam explosion, INTRODUTION Woerly waste, 1¢ wood chips, wood powder and bark, is a useful souree that ean be converted into fuel energy such at alcohol and methane". Though polyswcclaside (cellulewe wad Ieicelulose) in woxndy waste i a substrate for methane provision, diffi to convert the polysaccharide into methane by direst biological methods owing to lignin oovering the polysucchaside layers stimaly. Recely, slew explosion has heen adopted as one ofthe effective pretreatment methods for degrading and removing lignin from polysaccharide surface”. Tho. steam exploded ‘woody waste seems lo be converted into ruelhane, but the ‘yield of methee poxliced svight le conpauatively low because it has little nitrogen. It is generally known that not only carbon souree but abo nitrogen source are necessary forthe efficient methane fermentation”. On the collet Isl, chore is iteoyeassics sefeve of lala, Le seybean curd In Japan, processing industries discharge a3 much a3 £00,000 tons of okara anally. Thersfors, audition of okara into steam-exploded woody waste followed by fermentation seems Ube effective mcaan of sncreasing methane yiele MATERIAL AND METHOD Tn this shaly, the pretreatment effect af steam explosion on the methane production from woody waste ‘was examined using Japanese cedar as a sample. After the organic elemental analysis of okara and steam-exploded Japanese cedar, the okara was added into the steam- texploded Tapsiese cedar anal the mixture wis fecanented for mereasing the yreld of methane proceed Wood chips of Japanese cedar (Cryptomeria Japonica) were sed a a woody waste sample. The Tega wil widda of Uae word chips weve 2-3 sual 5-8 respectively Okara, +¢. soybean chrd resichie, was also used. The apparatus for steam explosion (Japan Chemical Engineering and Machinery Co. Ltd, Osaka, Japan) consisted of a slean generator, « pressurized reactor, a receiver ans] a condenser with a silencing action! The reactor was insulated to maintain a constant temperature The capacity of the reactor was 1.2 L, the highest pressure was 6 MPa and the highest temperature was 275, Approxinately 100 g of the chipped Tapauese coda was introduced into the reactor and then steam-heated at a steam pressure of 4.51 MPa anda steaming time of S min Total dry weight of steam-exploded Japanese cedar chips saad ekerer wats wljused ao sual whe ia 500 a conical flask containing S0X) ml, seed methanogens shudge tnd then ineubated aneerobieally for an incubation time of 700 h at 37°C. The percentages of okara in okara and steuni-exploded Jujauese cedar chips were set ws 0, 9,23, 50, 67 and 104P% The seed (methanogens shxige, 85 8000. ppm) was obtained by culturing the sludge ‘withdeawm from anaerobie digestion unit at a Kanazavra city wastewater treatent plant in Japan, Evolved gas was passed through a tube into 500 ml. cylinder that was submerged in saturated sodium chloride watcr. Stock gas ‘wae collected by a eyringe for analysis. The amounts of elements, C, Nand H, were determined by using an Corresponding Author: Yoshitoshi Nakamura, Graduate School of Natural Seionee and Technology. Kanazawa Univers Kalama machi, Kanazawa 920 1192, Japan, Tel: +81 76 234 4819 Fax: +81 76 234 4829 3354. Food Technol, ganic elements analyzer (CTIN CORDER MTS, Yamako Co Li, Kyoto, Japan) analysed by using a gas chromatcgraphy unit equipped with a thermal conductivity deteotor (GC SAPT, Shimazu Co Ltd., Kyoto, Japan), The column was stainless steel (4 by 03 an) filled with Shincarbon ST (Shimazu Co. Lud, Kyoto, Japan) and heated to 100°C: Helin gas (50-em° sin) was a canter gas 12 gas composition was RESULTS AND DISCUSSIONS Figure. 1 shows the time courses of total yas evolved sand uetlsane yas evelved i the metlsae fermentation of steamvexplaled Japanese cedar chips and untreated chips. The pl of reaction mixture, Le. methanogens sludge and sample was about 7 (data not shown), The amounts of total gas evolved and methane ga evolved increased with the increase of incubation time reaching eis unaxisutu values of 305 axl 180, respectively, at aan incubation time of 400}, On the other hand, ne gas evolved was detected in the methane fermentation of untreated chips. It was found that the steam explosion was a very effective pretreatment method for the enhancement of methane production from the Japanese ced chips, According wo Buswell’s Uaeory!, ubout 420 iL of melsaie gis prodvoal Gout 1 g of polysacclauide sich as cellulose anc hemicellulose. Since 1 @ of dry softwood contains about 0.7 g of polysaccharide!" the methane yield of holocellulose in the steam exploded Japanese cedar chips corresponded to approximately 0.61 ive, tlio of the auelhune gas peodueed, (180 mL) to Ue seovetial value, (294 auL). Table 1 shows the ameninis of CN, TT anel others in sample to the dry weight of sample, ic. steam-exploded Tapanese cedar chips and okara. Though litle difference ‘of carbon content between the steam-exploded Japanese cedar chips and the olara was observed, the mitrogen content rat of ekura was about 20 Ges lager Ua Wat OF the steameexplevesd Tapanese ceslar chips The: C/N ratio of steam-exploded wood chips was very high as 257, while the C/N ratio of okara was 12. The reason why the methane yield of steam-exploded Japanese cedar chips was Iow (Fig.1) seems to be that the steam-exploded Japanese cedar chips hve lle aitogen source, i, 0.22% On the other hand, since the akava is not a wood. based waste ul a soybean-processing waste, st has much nitrogen source. The composition of ohana is 25% protein, 19% lipid, $1% polysaccharide and 4% ash”, ‘Therefore, in order to inerease methane yield, it is necessary to add dhe okara into the steanexploded Tapsaese coda chips prio to fermentation process 3 (1): 535.537, 2005 {ble Omani anes ani of tam exploded Japanese hie michoa Stemvenoad 3639023923737 387 coli ok 469 406 10 ou op ‘able: Amount of alge and mene se lvl inthe meine ‘eee ting it pm Sage “otal Methane sas stean-erpoodeé ON fronedevohed vod chi haa) iio) aut) al) oo fo a0 1% on «30 z a 3 p ° 0 20304] 1ncbation ie (h) Fig. 1: Methane fermentation of steam-exploded Japanese cedar chips and untested chips. Syimbols, total gaa evolved of stcam-cxploded Japancac cedar chips, methane gae evolved of steam-exploded Tapuiese cedar chips, lolal gas evolved of ‘untreated chips methane gas evolved of untreated hips ‘Table 2 shows the methane fermentation results of sample containing steam-cxploded Japanese cedar chips and okara, Total dry weight of steam-exploded Japanese ced chi aad okara was adjusted to 1. Bod Uae total {gas and the methane gas evolved varied significantly with the C/N ratio. Though 1 g each of steam exploded Japanese cedar chips and okara produced 180 and 271 mL fof methane gas, respectively a larger amcunt of methane gas, ic, 315 mL, was obtaincd from 033 g of steam-exploded Fapanese cedar chips and 0.67 g of okara, 3364. Food Technol, 3 (1): 535.537, 2005 i.e CIN rationof 18. This result was che to the fact that in the low CON ratio, the growth of methanogens were restricted by the lack of nitrogen source while in the high COUN ratio, they were inhibited by ausuonia prodvced Grout large amounts of nitrogen source. A comparatively large amount methane gas is produced from only okara dium, ie, CIN of 12, but the effective methane pproskiction of unwdly waste reqnited the addition af nitrogen-tich material such as okara, This result also suggests thatthe addition of material with very high CT sul, ie. aint usave, night be awore effective fox Ue methane preduction of woody waste and this method can be also easily applied to various plant wastes such as wheat straw, bagusse and sweet sorghuan, Fuliae research will he focused on the more efficient methane gas production by mixing the different substrates and determining the optimal C”N ratio. CONCLUSIONS This study preseuted w uovel auelhod for inerewsigs the yield of methane gas proces from wooly waste ‘The steam explosion was used for the delignification of | woody waste and the large amount of methane as was Dprochiced from steam-exploded wordy waste while no methane gas was produced from untreated woody wast. Furthermore in order to increase the yield of methane gas de soybean residue, akara was added ead the additive elect was evaluated As a result, some valuable findings, ie. 1) the effectiveness of steam explosion as a pretreatment or methane fermentation of woody waste, 2) the adklitive effect of akara for increasing the yield of methane gas obtained from woody waste; 3) the ‘maximum C/N ratio for maximizing the amount of methane ‘gas produced, were obtained in this wouk ACKNOWLEDGEMENTS This study was supported i part hy a Granted for Scientific Research (B) (1) (No. 15360483) from the Ministry of Education, Culture, Sports, Science and Technology of Tapa, REFERENCES: 1. Klass, D.L., 1998. Biomass for Renewable Energy. Fuels wud Cheuiculs, Acudeuie Press, Sau Diego, CA, USA, Saddler. IN., 1993. Bioconversion of Forest and Agricultural Plant Residues, CAB. Intemational Press, Walling Fowl TK 3. Excofficr, G., E, Toussaint and V.R. 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