Mt s kt qu bc u trong vic s dng ch phm enzyme c cha pectinase trong ln men ht cacao Th.S Phan Thanh Bnh1 , PGS.

TS Nguyn Th Hoi Trm2, KS. Trn Th Hong Anh1, TS. Nguyn Vn Thng1, KS. H Vn Khm1, KS.Trn Th Thm H1, CN. Phm Vn Thao1, Th.S Hn Vn Trung3, Th.S Th Loan4, Th.S Ch Thnh5. Abstract Using pectinase in cocoa fermentation Using pectinase to reduce the mucilage of fresh cocoa beans and TA, increasing pH in cocoa beans is one of the ways of cocoa fermentation. Enzyme doses, days of fermentation, time of adding enzyme and number of turning during fermentation were done. Norms of Temperature, mucilage volume reducing, color beans, pH, TA, cup testing was collected and evaluated. The results show that: Using 80ppm enzyme dose (ppm/fresh cocoa beans) is better: temperature increased faster and higher (500C), brown beans 66.3%, purple beans 0.7%; pH =5.19, TA=1.26mgNaOH 0.1N/g; cup testing was better. Added pectinase when fermentation starting is better than other: temperature increased faster and higher (48.90C), brown beans 69.3%, purple beans 2.7%; pH =5.2, TA=1.16mgNaOH 0.1N/g; cup testing was better. Fermentation with 2 turns is better than other: temperature increased faster and higher (46.60C), brown beans 65.0%, purple beans 3.0%; pH =5.15, TA=1.28mgNaOH 0.1N/g; cup testing was better. Fermentation during 6 days are better than other: temperature increased faster and higher 49.20C), brown beans 73.0%, purple beans 1.7%; pH =5.27, TA=1.11mgNaOH 0.1N/g; cup testing was better. 1. t vn : Mt trong nhng cng on quan trng nht ca qu trnh s ch ht cacao cho sn phm t cht lng cao chnh l qu trnh ln men. Trong qu trnh ny c cht cho vi sinh vt v enzyme hot ng l ng trong lp cm nhy, tuy nhin ti lp cm nhy cn c cha mt lng cc cht khc nh nc, pectine, protein, cellulo trong pectine chnh l mt thnh phn c tc dng lin kt gia cc phn t li vi nhau to thnh th nhy (keo). Lng cm nhy ny c khi lng kh ln v ngn cn s pht sinh pht trin ca cc vi sinh vt, hot ng ca cc enzyme c trong . Mt kha cnh khc trong qu trnh ln men l khi c lng c cht ln th s to ra mt lng ln acid v t s cho sn phm c t l acid cao, pH thp (chua) nn sn phm t cht lng thp v khng c a chung khi pH di 5.
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Vin KHKT NLN Ty Nguyn Vin CN Thc phm 3 Trng Cao ng ngh TNDT Daklak 4 Trng Cao ng Lng thc, Thc phm H ni 5 Trng Cao ng Lng Thc, Thc phm Nng

Da trn nhng c s trn ni dung Nghin cu s dng ch phm enzyme pectinase b sung vo qu trnh ln men ht cacao c t ra nhm to ra c sn phm c cht lng cao, pH thp. 2. Ni dung v phng php nghin cu 2.1 Ni dung: - Xc nh liu lng, thi im b sung ch phm enzyme pectinase - Xc nh s ln o, thi gian ln men khi b sung ch phm enzyme pectinase 2.2 Phng php nghin cu: 2.2.1 B tr th nghim: Cc th nghim c b tr 1 yu t 3 ln lp li, c i chng trong tng th nghim trnh sai s do mi trng. TN1: Liu lng enzyme: cc cng thc 60ppm; 80ppm; 100ppm v i chng TN2: Thi im b sung enzyme: 0h, 24h, 48h v i chng TN3: S ln o: 1 ln o, 2 ln o, 3 ln o v i chng khng enzyme 2 ln o. TN4: Th nghim s ngy ln men: 4 ngy; 5 ngy; 6 ngy v i chng 6 ngy khng c enzyme. 2.2.2 Ch tiu v phng php theo di: - Tc gim ca hm lng cm nhy: % lng cm nhy/khi lng ht. - Nhit qu trnh ln men: o bng nhit k in t hin s Hana. - pH qu trnh ln men: o bng pH k bn Hana - TA (ml NaOH 0.1N/g) chun bng NaOH 0.1N pH 7 - nh gi mu sc ht bng quan st sau khi cuttest, tnh gi tr %. - nh gi cht lng th nm theo phng php ca AusAid. 3. Kt qu cc th nghim 3.1 Kt qu th nghim xc nh liu lng (nng CT1: 60ppm; CT2: 80ppm; CT3: 100ppm so vi trng lng ht ti): a/ S bin thin nhit Nhit l mt ch tiu xc nh mc ln men ca ht cacao. Kt qu biu 3.1 cho thy: nhit ca khi c s dng enzyme pectinase cao hn so vi i chng, tc tng nhit nhanh hn v c tnh n nh cao, iu ny l do qu trnh phn hy pectin lm tng nhanh mc thong kh v cc phn ng c s tham gia ca vi sinh vt hiu kh xy ra nhanh hn lm nhit trong tng cao. Trong cc cng thc th CT2 cho tin trin tt hn cc cng thc khc. th 3.1: S bin thin nhit trong khi ht 2

b/ S bin thin hm lng cm nhy Hm lng cm nhy gim mnh cc cng thc c s dng enzyme so vi i chng, sau 72-84h th mc gim c chm li v nm trong khong 7-9%, iu ny l do tc dng ca enzyme ch vi pectin nn cc c cht khc vn cn hin din trong qu trnh ln men lp cm nhy. Gia cc cng thc khng c s khc nhau r rt v tc gim hm lng cm nhy. th 3.2: S bin thin ca hm lng cm nhy c/ Kt qu nh gi mu sc bng phng php cuttest Cht lng mu sc ca cc cng thc cho thy s khc bit so vi i chng. T l ht nu cao hn i chng, t l ht tm thp hn (iu ny s lm tng cht lng ht cacao trong qu trnh bo qun). Trong cc cng thc th nghim, cng thc 2 (80ppm) cho kt qu tt hn vi 66,3% ht nu v ch 0,7% ht tm (ln men cha hon ton). Biu 3.3: Kt qu nh gi mu sc ht cacao d/ Kt qu xc nh pH v TA Kt qu xc dnh pH v TA cho thy cc cng thc u c pH cao hn (TA thp hn) so vi i chng. Gia cc cng thc khng c s khc bit nhiu(chnh nhau khong 0,04), mc d cng thc 2 vn cho pH cao nht. Biu 3.4: Kt qu xc nh pH v TA

e/ Kt qu nh gi th nm Kt qu nh gi th nm cho thy s khc nhau gia mu i chng v cc cng thc khng nhiu. Hng cacao ca CT3 l tt nht, v chua cng thc 1 cao nht. CT 2 (80ppm) vn cho kt qu trung bnh gia cc cng thc v c s hi ha, trong v chua c nh gi l thp nht (y l mc tiu ca vic s dng enzyme pectinase). Biu 3.5: Kt qu nh gi th nm cht lng ht cacao

3.2. Kt qu xc nh thi im s dng enzyme pectinase

a/ Kt qu xc nh nhit Kt qu nhit cho thy cc cng thc c tc tng nhit xut pht im nhanh hn v cao hn so vi i chng trong thi gian u ca qu trnh ln men. Gia cc cng thc khng c s khc bit nhiu. CT1 cho kt qu tng nhit nhanh hn cc cng thc khc.

th 3.6: Kt qu xc nh nhit trong qu trnh ln men b/ Kt qu xc nh hm lng cm nhy trong qu trnh ln men (thi im s dng E pectinase). Cc cng thc u cho hm lng cm nhy gim rt nhanh trong giai on u v sau 60- 72h th tc gim chm li. Tc gim ny ph hp vi cc tc ng ca enzyme. CT1 cho tc gim nhanh hn cc cng thc khc giai on u, sau 60h cc cng thc l tng ng. th 3.7: Bin thin hm lng cm nhy trong qu trnh ln men

c/ Kt qu xc nh mu sc ht bng cuttest Cc cng thc u cho s khc bit vi i chng, cng thc 2 (thi im b sau 24h) cho t l ht nu cao (71,7%) v t l ht c mu tm thp (1,7%). Cng thc i chng cho t l ht tm cao 5,7%, y l loi ht kh bin i tip tc mu sc trong qu trnh bo qun. Biu 3.8: Kt qu xc nh mu sc ht bng cuttest d/ Kt qu nh gi pH v TA Cc cng thc cho pH cao hn (TA thp hn) so vi i chng, cng thc 1 cho pH cao nht pH=5,2 so vi cc cng thc khc, tuy nhin s khc bit l khng nhiu. So vi cacao ca Ghana th gi tr pH thp hn nhiu (pH Ghana = 5,6-5,8)

Biu 3.9: Kt qu nh gi pH, TA e/ Kt qu th nm Kt qu th nm cho thy cc cng thc so vi i chng th khng c s khc bit r rng, so snh vi Ghana thi c s khc bit ln. Trong cc cng thc th CT1 cho kt qu kh quan hn cc cng thc khc. Mc hi ha, chua, cht, ng c gim hn so vi cc cng thc khc, c bit hng cacao th hin r hn. Biu 3.10: Kt qu nh gi th nm cc cng thc 5

3.3. Kt qu th nghim s ln o ca enzyme pectinase a/ Kt qu xc nh nhit Kt qu o nhit cho cho thy nhit tng kh u cc cng thc, i chng c thp hn cc cng thc nhng cng t nhit trn 450C. Cng thc 2 cho nhit nhnh hn cc cng thc khc t cao nht 49,20C 120 h. Trong thi gian ny nhit mi trng giao ng trong khong 27,9-31,20C nn qu trnh ln men cng din ra thun li hn.

th 3.11: Din bin nhit trong th nghim s ln o b/ Kt qu xc nh s bin thin hm lng cm nhy Hm lng cm nhy ca cc cng thc gim nhanh hn so vi i chng, iu ny ph hp vi tc ng ca enzyme vo lp cm nhy trong khi ht lm cho qu trnh thot dch cm nhy nhanh hn. Sau 60-72h th hm lng cm nhy c tc gim chm li v i chng bt kp vi tc gim ca cc cng thc.

th 3.12: S bin thin hm lng cm nhy trong qu trnh ln men

c/Kt qu xc nh mu sc ht 6

Mu sc ht kh cho thy mu tm ca cc cng thc c ci thin hn so vi i chng, cng thc 2 th mu nu c ci thin nhiu nht (65%) v mu nu tm cng thc 3 c gi tr cao nht. tit kim cng v chi ph chng ti chn s ln o l 2.

Biu 3.13: Kt qu xc nh mu sc ca ht kh d/ Kt qu xc nh pH, TA pH v TA ca cc cng thc khng cao hn i chng, tr cng thc 2, s chnh lch l khng ng k. Nh vy pH v TA khng nhng ph thuc vo qu trnh ln men m cn ph thuc vo hm lng cm nhy, loi ging, v cc iu kin khc.

Biu 3.14: Kt qu nh gi pH v TA ca ht kh e/ Kt qu xc nh cht lng th nm Cht lng th nm cho thy cng thc 2 c mc hi ha gia cc ch tiu hn cc cng thc khc. Mc d v ng cng thc 1 cho gi tr thp hn, v chua cng thc i chng cho gi tr thp hn. Nh vy vi 2 ln o cho ta gi tr th nm c mc hi ha cao hn cc cng thc khc. Biu 3.15: Kt qu xc nh cht lng th nm 4/ Kt qu th nghim thi gian ln men enzyme pectinase 7

a/ Kt qu xc nh nhit Nhit trong qu trnh ln men din bin khng t yu cu, sau 48h nhit ch t khong 400C, thm ch i chng cn ch t 37,80C. Qu trnh ln men chu nh hng ca nhit mi trng (Ch giao ng trong khong 20-240C).

Biu 3.20: Nhit trong qu trnh ln men th nghim thi gian ln men b/ Kt qu xc nh s bin thin hm lng cm nhy Kt qu xc nh hm lng cm nhy cho thy qu trnh ln men c s dng enzyme c tc thot cm nhy tt hn so vi i chng, cc cng thc khc nhau th khng c s khc nhau. Thi gian ln men di ch lm gim mt phn lng cm nhy nh, t nh hng ti cc kt qu v s hao ht khi lng ca ht. Bng 3.21: Kt qu xc nh bin thin hm lng cm nhy c/Kt qu xc nh mu sc ht Kt qu mu sc ht cho thy mu nu cao ln theo thi gian ln men v mu tm thp dn theo thi gian ln men. Cc cng thc u cho mu tm thp hn nhiu so vi i chng (1,7% so vi 11,7%) y l loi ht kh chuyn thnh mu mu trong qu trnh bo qun.

Biu 3.24: Kt qu nh gi mu sc ht cacao d/ Kt qu xc nh pH, TA Kt qu cho thy pH tng theo s ngy ln men, TA gim theo s ngy ln men. Ln men c s dng enzyme 6 8

ngy vn cho pH cao v TA thp hn so vi cc cng thc khc v so vi i chng (5,09; 1,25). Nh vy chng t trong qu trnh ln men ngoi sn sinh nhiu acid th cn c qu trnh chuyn ha khc v qu trnh bay hi ca cc acid c th bay hi. Biu 3.25: Kt qu nh gi pH v TA e/ Kt qu xc nh cht lng th nm Kt qu biu 2.26 cho thy cht lng v hng v cng tt khi s ngy ln men cng nhiu (trong thi gian 6 ngy). V chua gim xung, v ng v cht cng c ci thin ng k. Hng tri cy th din bin ngc li, iu ny c th do qu trnh phn hy xy ra khi ln men di ngy.

Biu 2.26: Kt qu xc nh cht lng th nm Kt lun: Vi cc kt qu ca th nghim th vic s dng enzyme pectinase s lm tng cht lng trong mu sc, v chua (pH, TA) l 2 ch tiu c ci thin ng k. iu ny gip cho sn phm cacao Vit Nam c v chua thp hn hin ti. T cc kt qu th nghim cho thy chng ta c th s dng enzyme nh sau: - Nng 80ppm khi s dng ch phm enzyme pectinase (Ultrazyme AFPL) - Thi im x l l ngay t lc bt u ln men do c pH cao, kt qu th nm tt, kt qu mu sc ht khng km nhiu so vi cc cng thc khc - S ln o l 2 ln trong sut qu trnh ln men. - Thi gian ln men l 6 ngy khi ln men khng s dng thit b.