TAP CHi KHOA HQC & CONV, NGHE CAC TRlfOTVG D^l HOC K\ FHIAT * SO 98 - 2014 TIEN XIJ' LY ROM RA BANG AXIT AXETIC B SUNG AXIT CLOHYDRIC VA DlTCJNG HOA BANG ENZYME CHO SAN XUAT ETANOL SINH HOC ACETIC ACID WITH ADDED HYDROCHLORIC ACID PRETREATMENT A N D ENZYMATIC HYDROLYSIS OF RICE STRAW FOR BIOETHANOL PRODUCTION Nguyen Thi Minh Phirffng, Le Quang Dien, Doan Thai Hoa Trudng Dgi hoc Bdch khoa Hd Ndi D i n Toa soan 26-10-2012, chdp nhdn ddng 25-12-2013 TOM TAT Bay bao trinh bay kit qua nghien ciru tiin xw ly ram bang axit axetic bo sung axit elohydrie va dwdng hda bing enzyme cho san xuat etanol sinh hoe Sw thay ddi eau triic nguyen lieu qua trinh xw ly cung da dwgc nghien eiru bang ky thuat hien vi dien tir quet Da xac dinh duxyc che dd edng nghe thich hgp thu nhan dwdng khCr tir ram ra- tiin xd' ly bing axit axetic bS sung 2% (vi thi tich) axit elohydrie, a 100°C 90 phut, ti dich xw ly 1.10, ket hgp thuy phan bing enzyme cellulase Celtic® CTec2 eua hang Novozymes V&I hoat Iwe 301,3 FPU/ml), cho ham Iwgngdwdng khw dat 38-39% so vai nguyen lieu khd tuyet doi (KTD) O diiu kien xd ly eo tdi gin 60% nguyen lieu bi chuyen hda twang img gin 90% lignin gin 60% cac chit vd ea bi hda tan Ngoai tignin cd anh hwang quyit dmh tdi hieu qua dwdng hda xenluloza Td khoa Sinh khoi, etanol sinh hoc, tien xie ly, axetic axit, dudng hda being enzyme ABSTRACT This paper presents results of study on rice straw pretreatment by acetic acid with added hydrochloric acid and enzymatic saccharijication for bioelhanol production The sirucliiral changes of rice straw after treatment has been studied by scanning electronic micro.fcopy (SEM) The appropriate technology conditions for obtaining of reducing sugar from straw was determined as pretreatment by acetic acid -with adding of 2% vol hydrochloric acid, al 100"C for 90 minutes, liquire ratio 1:10 and combined enzymatic hydrolysis by Novozymes cellulase Cellic® CTec2 with activity of 301.3 FPU/ ml given yield of reducing sugar over 38-39% of over dry rice straw In these tretreatment conditions about 60% of Ihe /•OM' material has been conversed, 90% of lignin and about 60% of dissolved inorganic substances have been degraled In additon lignin has a decisive influence in cellulose saccharificalion Keywords: Biomass, bioethanol, rice straw, pretreatment, acetic acid, saccharification M O DAD M , , • J ,- V, • I' J •- -7 c Nuoc ta CO dien tich lua dat gan 7,5 ,•-, L • • I ]• ui', ' AC trieu va san lugng lua hang nam dat tren 45 irilu tdn [ I ], Udc tinh co khodng gan 50 trieu tdn rom phi tlia, sau thu hoach cjn xu iy La p h i phu pham n6ng nghiep chira xenluloza va lignin -x -^u-*- J I • i ?-i tiem nang co the tan dung cho san xuat nhieu s a n p h a m c o g i d t r i nhu hot gidy nhien lieu sinh hoc hda chat,,,, nhung hien mdi chi mot ,•• uJ • J ,• L- L - ' phan nho ram duac su dung am phan bon, u-.A' -.!•u " , , •chat dot vat heu phuc vu cone, nana nghiep I • u- A', K- - I- • i- • \.u wai b dot bo,_gay lang phi va anh huong nghiem trgng den moi truong Nghien ciiu liim ndng va tinh chdt ciia rom [2] cho thdy, day Id nguon nguyln lieu phii hop cho san xuat etanol sinh hgc Tren the gidi, nhieu nghiln cuu cdng nghe tien xu ly rom bdng cac tac nhan khde nhu no hoi co •, ^ , , , j , , , -, -, xuc tac, nuoc nong ket hop vol kiem, tien xu y i ° '•- 11, • • ^~ A A bang vi song da dugc triln khai vd da dat dugc ''^l ' ' " \ " ! ' ^ ' ^}"*' p i " ! ? ' Tuy "hien, theo danh f : "Shuj" cini cdc cong nghe kha thi van la huong nghien CUU can duoc tiep tuc ° ^ •• '•H 'y-2 VAT LIEU VA P H f O N G PHAP n„ J L, I •- Roni su dung cho nghien ciru dugc thu ,, - u •• k , , x , - r, , ; •• gom o huyen Hoai Due, Ha Noi Rom kho gio ^ u u„• u-; : •^^^^'^ nghien bang may nghien biia vd sdng o ^^^ ^^_ ^^^^ , ^ ^^.^ ^^^^ ^ p,^.^ ^oa hgc c a ban cua rani duge xac djnh theo cac tilu chudn TAPPl d6i vdi nguyen lieu thuc vat TAP CHi KHOA HQC & CONG NGHf CAC TRUONG DAI HOC KY THIAT * SO 98 - 2014 Qua trinh tiin xu ly ram dugc tiln hanh binh thiiy tinh 500 ml, ldp vdi sinh hdn ngugc gia nbiet bdng bl glyxerin Mdi me tien hanh vdi khoang 20 g nguyln Heu khd gid Thdnh phdn dich ndu (CH3COOH, HCI), nhiet 66 vd thdi gian xu ly dugc dilu chinh tiiy theo muc tilu ciia tdng thuc nghiem Tl djch xd ly (khoi lugng nguyen lieu khd tuyet doi/ the tich djch ndu) cd djnh la l/IO Bdt thu dugc sau xuly dugc danh toi, rua bdng axit axetic, dung dich NaOH vd nude nong, vdt nude va tilp tuc dugc thiiy phdn bang enzyme thuang pham Cellic® CTec2 ciia hang Novozymes d 50°C 120 gid, su dung dem axit citric co pH 4,8^5.0, vdi n6ng dp hot 10% Hdm lugng dudng khu dugc xac dinh theo phuang phap DNS [6], bang may quang Optima-sp 300 KET QUA VA BAN LUAN 3.1 Thanh phan hda hgc co' ban ctia nguyen li^u Thanh phdn hda hgc ca ban cua nguyln lieu dd xdc dinh theo cdc phuang dugc trinh bdy tren Bang 1, rdng, axit axetic Id dung mdi co khd ndng hda tan lignin (thdp phan tu), ddc biet la a nhiet d6 cao Qua trinh ndy se dugc thiic ddy sd dung axit axetic hdn hgp vdi mgt sd hgp chat khac, nhu axit sunfuric, axit elohydrie, hydropeoxit nhu'ng dilu kiln nhdt djnh, cac hgp chdt ndy cd kha nang tao vdi axit axetic mgt tdc nhdn cd kha ndng phan huy lignin cao phan tu cac hgp chat thdp phdn tu, de ddng hda tan dung djch axit axetic Trong nghiln cdu nay, HCI dd dugc lira chgn su dung, Co chi hda hgc tach loai lignin bang hdn hgp dugc thuc hien bang su phdn hiiy lignin bdi axit cloaxetic, t^o td axit axetic vd axh elohydrie Xir ly ram r? theo phuang phdp neu trin, dugc tiln hdnh d nhiet lOOX, 60 phut, vdi ti dich 1:10 Cdc thdng so ndy duoc lira chgn bang mgt loat cdc nghiln cim sa bg Ham lugng (thi tich) HCI hdn hgp vdi CHjCOOH dugc chgn khodng 0-4% Bdt xenluloza thu dugc sau dd dugc thiiy phdn bang enzyme theo phuong phdp da trinh bay tren Bdng Thdnh phdn hda hoc ciia rom rg TT Thanh phan Xenluloza (TAPPI T207 cm99) Holoxenluloza (TAPPI T 249) Pentozan(GOST 10820-75) Lignin (TAPPI T222 om-98) Bo tro (TAPPI T211 om-93) Ham luong (%) 40,2 62,1 18,7 21,5 14,7 3.1 Anh hudng cua cdc yeu Id cdng nghe lien xir ly ram rg tdi hieu qud dudng hda bdng enzyme Muc tilu ciia nghiln cuu la dat dugc mirc dudng hda xenluloza cao nhdt vdi mgt che cong nghe thich hgp nhdt, Chi tilu danh gia hieu qua dudng hda xenluloza Id lugng dudng khir so vdi nguyln li?u ban ddu, Ngoai ra, thdnh phdn hda hgc cita bd rdn cdn lai sau thuy phdn bdng enzyme cung dd dugc phdn tich, de ddnh gid mdc chuyen hda ciia xenluloza 3.1.1 Anh hudng cita axit elohydrie Cac nghien cdu ve ca che hda hgc cua qua tr • •-[ tdch loai lignin bdng axit hO^ ca [4] da chi Nong axit HCI (%) Hinh Anh huang ciia ndng axil HCI den su chuyen hda nguyen lieu vd hdm lugng dudng khu (l:Lirgng nguyen lieu hoa lan 2- Hdm lug-ng dirdng khir so vai nguyen lieu KTD) Td kit qua thu dugc (Hinh I) cd thi thdy, trudng hgp xu ly ram bdng axil axetic khdng bd sung HCI, c6 khoang 20-22% khoi lugng nguyln lieu hda tan, c6 mdt lugng nhd lignin, Tuy nhien, cho dii cdu tao ciia nguyen lieu da bj thay ddi song qua trinh dudng hda xenluloza dd khdng dien Khi bd sung HCI ciing vdi tdng bam lugng, qua trinh tdch TAP CHi KHOA HOC & CONG NGHE CAC TRl!J khep kin, it chdt thai, ddy Id uu diem ldn ciia phuang phap tien xu ly ram bdng axit axetic c6 bo sung xiic tdc HCI so vdi cac phuang phap tien xir Iy khac TAI LIEU THAM KHAO Nien giam thong kl ndm 2011 , L,Q Dien, D.T Hoa, N.T.M, Phuong, N,T.M Nguyet Rice straw and corn stalk in Northern Viet Nam as potential lignocellulosic sourse for production of bioethanol and other value added product, Proceeding of 8"' Biomass - Asia Workshop, Hanoi, Vietnam; 20 i 1; 82 Binod P, Sindhu R Singhania RR, Vikram S, Devi L Nagalakshmi S, Kurien N, Sukumaran RK, Pandey A.: Bioethanol production from rice straw: An overview, Bioresource Technology, Vol, 101 N13,(2010) , Anwar, Z., Gulfraz M., Imran M., Asad, M.J., Shafi A L, Anwar P and Qureshi R Optimization of dilute acid pretreatment using response Surface Methodology for Bioethanol Production from Cellulosic Biomass of Rice Polish, Pak J Bot 44(1): 169-176 (2012) , Esa Murinen, Organosolv pulping, Oulun Ypiopisto Oulu (2000) , PCS TS LI Thanh Mai GS TS Nguyln Thi Hiln PGS TS Pham Thu Thuy, TS Nguyln Thanh Hdng, ThS LI Thi Lan Chi, Cdc phirong phap phan tich nganh cdng nghe Ien men NXB KH&Kl, l-la Npi (2007) Nguyen Thi Minh Phuong -Tel: 01216143493 Email: phuong.nguyenthiminh@hust.edu.vn Vien Ky thudt Hoa hgc, Trudng Dai hgc Bach khoa Hd Ndi S6 I - Dai Cd Viet - Hai Ba Trung - Ha Noi ... ldn , ban, tao dilu kien tdt cho enzyme tilp can vd thuy phan xenluloza KET LUAIN -Tiin xu ly ram bdng axit axetic co bd sung HCI kit hgp dudng hoa bang enzyme cho hdm lugngdudng khu dat 38-39%... djch axit axetic Trong nghiln cdu nay, HCI dd dugc lira chgn su dung, Co chi hda hgc tach loai lignin bang hdn hgp dugc thuc hien bang su phdn hiiy lignin bdi axit cloaxetic, t^o td axit axetic. .. rdng, axit axetic Id dung mdi co khd ndng hda tan lignin (thdp phan tu), ddc biet la a nhiet d6 cao Qua trinh ndy se dugc thiic ddy sd dung axit axetic hdn hgp vdi mgt sd hgp chat khac, nhu axit