KHOA HOC - CONG NGHE - MOI
TRUCJNGXAC DjNH LOAI ENZYME PHU HQlP DE T A C H LOP NHdiT CUA HAT CA PHE TRONG CHE BIEN THEO
PHlTOfNG P H A P UfiT
LDATVANDE
Mdt edng doan quan trgng trong ehe bien e i phe nhin theo phuong phip udt l i tich Idp nhdt nim tren vd thdc sau khi xit bd vd qui. Hai phuong phip tich nhdt phd bien hien nay Ii len men tu nhien v i dinh nhdt co hgc. Mdi phuang phip deu ed ban che nhit djnh:
Len men ty nhien cin xiy dung nhieu be len men tdn kem v i ehju nhieu tie ddng khdng kiem soit duge tir ben ngoii, nhit li yeu td nhiet do, do dd ket qui thu duge khdng dn djnh. Chit lugng ci phe cd nguy co b j giim do nhieu hat bi len men thdi din
T.S Nguyen Van Thitdng Vien KHKT Ndng ldm nghiep Tdy Nguyen
den eic mui vj xiu trong nude pha. Len men tu nhien lim ting miie do d nhiem nude v i khdng khi.
Phuong phap danh nhdt co hoc khac phue dugc mdt vii bit lgi cua phuang phip len men tu nhien, nhung nd lai ed eic bit lgi khie: thiet bj ed gii khi eao; tieu tdn nhieu dien; nguy co Iim nhieu hat bj trdc vd thdc v i e i phe nhin khdng edn ldp bao ve nen khd bio quin. Do eic miy dinh nhdt khdng tieh duge nhdt nim tai khe hat nen mudn loai bd bet nhdt phii len men bd sung, v i khi dd qui trinh ehe bien trd nen phiic tap ban.
H.L Ulin men cdphi H.2. Ddnh nhat (Anh: VT)
De trinh duge nhiing ban che trong c i hai phuong phip len men tu nhien v i dinh nhdt co bgc, cae nhi khoa bgc de nghj sir dung enzyme de phin buy ldp nhdt. Enzyme khdng phii Ii hda chit m i l i chit xuc tie sinh hge, ed trong te bio eiia mgi loai sinh vit. Nd ed bin chat protein, ed tinh xiic tic die
H.3. Mdt logi chi phdm enzyme ca hgc ddng can 30 lit sie dung cho cd phi
(Anh: VT) (Anh: VT) hieu v i lim eho eic phan iing hda hge xiy ra cue ky nhanh. Cie hing sin xuit enzyme hien nay ehu yeu tich triet enzyme tir vi sinh vit.
Trudc khi iing dung enzyme de phin buy ldp nhdt, diiu cdn thiit ddu tiin la phdi xdc dinh dugc logi chi phdm enzyme phii hgp vai logi ca chat
BAN TIN THONG TIN KH&CN SO 01/2011
KHOA HOC - CONG NGHE - MOI
TRJCJNGtrong lap nhat. Thinh phin chinh cua Idp nhdt Ii cac hgp chit eao phin tir nhu pectin, cellulose vi cie loai dudng. Dudng tan trong nudc nhung pectin vi cellulose khdng tan trong nudc. Vi vay trong cic thi nghiem, ehung tdi chi sir dung cie ehe pham xiic tic chiia enzyme peetinase hoac cellulase, hoac chira hdn hgp eic enzyme thude hai nhdm niy. Mue tieu nghien ciru nhim xie djnh loai che phim enzyme phu hgp de phin buy ldp nhdt cua hat ci phe vdi (Robusta) trong phuang phip ehe bien udt qua viee dinh gia hieu lue phin huy dich chiet ldp nhdt eua ehung.
2. PHUONG PHAP NGHIEN CUtJ 2.1. Quy trinh chuan hi mau
- Qui chin ci phe vdi (thu hoaeh thing 12 nim 2009 tai cic vudn ciy d Budn Ma Thudt) dugc tich vd bing miy xit trdng thdng thudng de lay ra 30kg ciphe thdc.
- Ci phe thdc dugc chia lim 6 phin, mdi phan 5kg, dung trong eic chiu i-nox; cho them vio mdi chiu 2,5 lit nudc (theo ti le 2 ei phe/1 nudc). Dung miy dinh trimg cd hai cinh khuiy de khuay dio ky ei phe d mdi chiu trong thdi gian 10 phut de chiet
xuat ldp nhdt. ; - Tich rieng hat ca phe ra; trdn chung djch
chiet cua 6 chau de ed miu ehung, sau dd lai ehia deu eho 6 chiu de ed miu thi nghiem xir ly enzyme.
Bang 1. Cac che pham enzyme thyc pham su* dung trong nghien cihi Che pham
Lumipect-CP
Peelzym®
Rohapect®
lOL
Rohament®
pulpex
Pectinex®
Ultra SP-L
Thanh phan enzyme Polygalacturonase (viet tit PG, thudc nhdm peetinase)
Enzyme nhdm cellulase, chu yeu ji-glucanase;
ngoii ra cdn cd xylanase vi hemicellulase nhung boat luc thap
Enzyme nhom peetinase, gom pectinesterase (PE), VG xk pectinlyase (PL) Enzyme dhbm. peetinase gdm PG, PE, PL; ngoai ra cdn cd mdt sd enzyme nhdm cellulase nhung boat tinh thap
PG, ngoai ra cdn cd hemicelllulase nhung boat tinh rit thip
Nguon goc
Sin pham eua Lumis Biotech P.Ltd,, An Do Sin pham cua Novo Nordisk Ferment Ltd, Dan Mach. Sin xuit bang gidng nim Aspergillus aculeatus.
Sin phim cua AB Enzymes, Phin Lan.
San xuit tir gidng nim Aspergillus niger.
Sin phim cua AB Enzymes, Phan Lan.
San xuat tir gidng nam Aspergillus niger.
Sin phim ciia Novo Nordisk Ferment Ltd, Dan Mach. Sin xuit tir gidng nim Aspergillus aculeatus.
Tinh chat Ldng, mau nau, tan trong nudc. Hoat luc tdi thilu: 720 LU/ml.
Ti trgng: l,25g/ml.
Ldng, miu nau, mui len men nhe, tan trong nudc. Hoat luc tdi thilu: lOOFBG/g. Ti trgng: l,23g/ml Ldng, miu nau va mui thgm, tan trong nudc.
Hoat luc tdi thieu: 260 PA. Ti trgng: l,18g/ml.
Bdt thd, miu nau nhat, tan trong nudc. Hoat luc tdi thilu: 600 PA.
Ldng, mau nau, miii len men nhe, tan trong nudc. Hoat luc: 9500 PGU/ml. Ti trgng:
l,I6g/ml
BAN TIN THONG TIN KH&CN SO 01/2011
KHOA HOC - CONG NGHE - MOI TRLfCJNG
- Chuin bj 5 ehl phim enzyme (neu d bing 1), mdi loai enzyme xir ly djch nhdt d mdt chiu rieng biet vdi lilu lugng 20 phin trieu (ppm) tinh theo khdi lugng e i phe thdc udt. Djch chiet trong chiu edn lai li ddi chiing (khdng xii Iy enzyme).
2.2. Danh gia hieu lire ciia enzyme
Sir dung hai loai vi nhdt ke VT-03F v i VT- 04F eiia Nhit Bin de do do nhdt eua djch chiet ngay tai thdi diem trudc khi xir ly enzyme v i sau mdi 6
phiit. Sd lin do eho mdi edng thire xir Iy enzyme Ii 15 lin. Don vj do nhdt tinh bing mPa.s (Millipaseal-giiy). Miie do giim do nhdt nhanh hay ehim thi hien hieu lue cua enzyme eao hay thip.
3. KET QUA VA THAO LUAN
Ket qui trong bing 2 eho thiy c i nim loai enzyme deu cd tie dung lim giim nhanh do nhdt eua djch triet Idp nhdt cua hat ei phe.
Bang Thdi gian
xii' ly (phut)
0 6 12 18 24 30 36 42 48 54 60 66 72 78 84
2. D o nhdt (dvt: mPa.s) cua dich chiet ldp nhdt ca phe sau khi xir ly enzyme Cac cong thirc xw ly enzyme
Lumipeet - C P
270 230 185 155 110 95 80 70 65 60 55 50 50 45 28
Peelzym®
270 60 50 20 17 15 13 10 10 10 10 8 8 7 7
Rohapect®
lOL
270 70 60 35 25 17 16 15 14 13 11 10 12 10 10
Rohament®
Pulpex
270 155 118 60 40 17 9 6 5 5 5 5 4 4 4
Pectinex®
Ulha SP-L
270 140 100 65 29 26 23 20 19 17 17 16 16 16 16
Ddi chirng
270 270 270 265 265 260 260 260 250 250 250 250 250 250 250
Sd lieu cbi ra ring, do nhdt cua djch chiet ldp nhdt trudc khi xir ly enzyme l i 270 mPa.s; sau khi xir ly, Peelzym® lim giim do nhdt nhanh nhit (sau 18 phut do nhdt djch chiet cdn 20 mPa.s) v i tilp theo li Rohapect® IOL (sau 24 phut do nhdt djch chiet cdn 25 mPa.s).
Ciing thiy ring Rohament® Pulpex d giai
doan diu khdng lim giim do nhdt nhanh nhu Peelzym® v i Rohapect® lOL, nhung d giai doan sau lim do nhdt cua djch chiet giim triet de nhit:
sau 48 phut, do nhdt djch chiet cdn 5 mPa.s, chi bing 1/2 so vdi djch chiet xir ly Peelzym® v i bing 1/3 so vdi djch chiet xir ly Rohapect® I OL. Luu y ring do nhdt cua nude (lay tir gieng khoan) sir dung trong thi nghiem nay la 0,7 mPa.s.
BAN TIN THONG TIN KH&CN SO 01/2011
KHOA HOC - CONG NGHE - MOI
TRUCJNGHinh 4 bilu diln qui trinh giim do nhdt cua dich chiit bing sd tuong ddi (%). Trong khi ddi ehiing khdng xir ly enzyme, sau 3 0 phut do nhdt van cdn 94,7% thi cic edng thiie xir ly enzyme, do nhdt chi cdn 5-33%. Sau 84 phut, do nhdt cua ddi ehiing van cdn 87,7%, trong khi do nhdt cua cic cdng thiie xir ly ehi cdn dudi 10%).
Trong 5 ehl phim, Lumipeet CP ed hieu luc thap (sau 30 phiit do nhdt van cdn 30%o). Cdn lai 4 enzyme khic ed hieu lue tdt, sau 30 phut lim do nhdt dich ehiit giim edn 5-10%; tuy viy trong 4 loai niy, Pectinex® Ultra SP-L eho hieu lue kem hon ba loai edn lai.
120.0
100.0
80.0
60.0
40.0 - 20.0
- L\imipect- CP - Peelzym®
-Rohapect® lOL - Rohament® Pulpex - Pectinex® Ultra SP-L - Doi chung Dich nhot - Doi chiing Nuac
0 6 12 18 24 30 36 42 48 54 60 66 72 78 84 Thoi gian xii ly (phut)
H.4. Dd thi bieu dien miec do gidm do nhdt tuang ddi cua dich triet ldp nhdt cd phe sau khi xie ly bang cdc logi enzyme
Che phim Peelzym® cd hieu luc rit eao, Iam giim do nhdt eiia dich triet ldp nhdt nhanh nhit: ehi sau 18 phut xir ly, do nhdt eua djch triet ldp nhdt ehi eon 7%. Rohapect® lOL cho cdng ed hieu lire rit eao; sau 24 phut xir ly, do nhdt cua djch chiet cdn dudi 10%,.
Rohament® Pulpex eiing eho bieu lue cao.
Theo edng bd, boat lue peetinase eua ehe pham niy li 600 PA, eao ban 2 lin so vdi Rohapect® I OL (260 PA); tuy viy ket qui nghien cim lai cho thiy hieu lire phin buy djch nhdt cua Rohapect® lOL eao hon. Cd le do bio ehe d dang bdt nen Rohament®
Pulpex khd hda tan vio djch nhdt hon, vi viy phit huy tie dung ehim hon. Vdi uu tien lua chgn che phim lim giim do nhdt dich triet xudng dudi 10%) nhanh nhit, nen Rohament® Pulpex khdng dugc ehgn cho eic nghien cim tiep theo.
4. KET LUAN VA KIEN NGHI 4.1. Ket luan
- Trong cie ehe phim enzyme peetinase v i cellulase, ehe phim chira hdn hgp nhieu loai
enzyme cd hieu luc cao ban che pham chi chiia enzyme don le.
- Hai ehe phim Peelzym® (hdn hgp eic enzyme nhdm cellulose) v i Rohapect® lOL (hdn hgp cic enzyme rdiovti peetinase) cd hieu luc phan huy djch ehiet ldp nhdt eua qui ea phe eao nhit.
4.2. Kien nghi
Nghien ciiu xie djnh dieu kien tdi thich ve lieu lugng, pH v i nhiet do mdi trudng de hai che phim Peelzym® v i Rohapect® lOL phan buy ldp nhdt c i phe tdt nhat./.
TAI LIEU THAM KHAO CHINH 1. Carlos H. J. Brando (2004), Harvesting and Green Coffee Processing, in Coffee Growing, Processing, Sustainable Production, Wiley-VCH Veriag Gmbh & Co. KgaA, Gennany 605-714 2. Tai lieu ky thuat (2009) eua cie hang AB Enzyme, Novozyme va Greentech.
3.Reinhold Urlaub (2008), Enzyme application in coffee processing, R-D Reports of AB
Enzymes, EDSTACHEM (M) SDN, BHD.