TAP CH PHAT TRIEN KH&CN, TAP 11, SO 12 - 2008

NGHIN CU CH TO CH PHM BIOCOFFEE-1 T ASPERGILLUS NIGER V NG DNG LN MEN CC LOI C PH

L Hng Ph(1), Nguyn c Lng(2), i Ngha(3) (1) Trng i hc Quc T, HQG -HCM (2)Trng i hc Bch Khoa, HQG -HCM (3)Trng i hc Nng Lm Tp.HCM
(Bi nhn ngy 30 thng 01 nm 2008, hon chnh sa cha ngy 18 thng 08 nm 2008)

TM TT: Trong ch bin c ph hin nay, vn tch chit hu ht cc cht ha tan c trong ht c ph c cc nh sn xut quan tm. Tuy nhin, qu trnh tch chit cc cht ha tan c trong ht c ph gp tr ngi do hai thnh phn pectin v cellulose chim ch yu trong ht c ph gy ra, pectin (52,62-55,14%), cellulose (15,29-17,04%). Chng ti ch to thnh cng ch phm biocoffee-1 cha hot tnh pectinase v cellulase cao vi thnh phn mi trng ti u l 9% c rt, 15% v tru v nhng thnh phn khc l 75% cm v 1% NH4(SO4)2 v cc yu t khc nh sau: iu kin nhit khong 27-300C (nhit phng), m t 56-64%W v thi gian t 40-44 gi. Qu trnh nhn ln ca ch phm, hai loi bt m v bt sn nhn thy l tt nht v t l tt nht l bt m: bt sn = 2:1, vi m 56% v thi gian 30 gi. Ngoi ra, chng ti xc nh cc iu kin ti u cho qu trnh ln men c ph l c ph nn ngm nc 1 gi v ro, ln men 16 gi v t l ch phm: trng lng c ph s dng l 16/1000. Ch phm ny va gii quyt hiu qu tr ngi c cp t u ng thi nng cao cht lng c ph bng cng ngh ln men, mt hng mi trong ch bin c ph hin nay.[3], [4], [5] 1.GII THIU C ph l cy cng nghip quan trng v c gi tr kinh t i vi cc quc gia nhit i, trong c Vit Nam. Hin nay c ph trong nc c gi tr xut khu ng hng th hai ch sau la go. C ph c s dng rng ri n mc ph thng v khng ch c ng dng trong ngnh cng nghip thc ung m cn c ng dng trong cc cng ty bnh ko, thm ch trong cc cng ty dc phm. Trong cc ngnh ny hu nh sn phm cha c ph c s dng dng ha tan.[1]. Tuy nhin, hin nay vn trch ly cc cht ha tan c trong ht c ph gp tr ngi ln nu s dng phng php thng thng. C ngha l mt lng ln cc cht ha tan vn cn nm li trong b sau qu trnh tch chit. Trong nhiu nghin cu cho thy rng, s cn tr ny ch yu do hai thnh phn pectin v cellulose c trong c ph gy nn. Hai thnh phn ny l hai thnh phn chim t l ln v c cu trc sinh hc kh bn vng v chng vn l lp che ch t bo v cht lin kt t bo khi tri c ph cn sng [5]. Nhn thy cu trc ny khi trng thi mt nc cng tr nn bn vng, chng ch c th b ph hy khi dng cc tc nhn axit, baz mnh m nhng tc nhn ny khng c khuyn khch trong ch bin thc phm ngy nay [4], [5]. gii quyt vn ny, chng ti nhn thy c th ng dng cc ch phm vi sinh vt c hot tnh pectinase v cellulase tc ng mt cch c hiu ln c cht pectin v cellulose. Ngy nay vic hng n s dng ch phm vi sinh vt trong ch bin l hng i mi, hiu qu v nng cao cht lng c ph v ng thi tng kh nng trch cc cht ha tan.[1], [3], [4]. Hin nay Vit Nam c 3 lai c ph c trng kh ph bin l c ph vi (C. robusta), ch (C. arabica) v mt (C. excelsa). Sn phm c ph c bn trn th trng hin nay hu ht c s dng phng php truyn thng (phng php kh) cho c ph cht lng trung bnh. Trn th gii mt phng php mi ang c s quan tm ca cc nh ch bin c ph,

Trang 53

Science & Technology Development, Vol 11, No.12 - 2008 l phng php ch bin t, cho c ph cht lng cao [5]. Nghin cu ny gp phn vo vic nng cao cht lng c ph Vit Nam p ng nhu cu thc tin xut khu c ph. c bit song song vi vic nng cao cht lng c ph, chng ti cng trch ly hu ht cc cht ha tan c trong ht nhn c ph. Bi bo ny chng ti tp trung vo 4 mc tiu ln nh sau: 1. Phn tch cc thnh phn c trong nhn c ph 2. Ti u ha cc iu kin sn xut ch phm vi sinh vt sinh tng hp pectinase v cellulase 3. Kho st s dng mt vi dng tinh bt trong nhn ging ch phm vi sinh 4. Kho st ti u ha iu kin ln men 2.VT LIU V PHNG PHP NGHIN CU 2.1.Vt liu Chng vi sinh vt Aspergillus niger c nhiu kho st c kh nng sinh tng hp enzyme cellulase cao (trong mt ti tin s) v c kh nng sinh tng hp pectinase cao (mt ti thc s). Chng vi sinh vt ny c cung cp t phng th nghim Cng ngh Sinh hc, H Bch Khoa TP.HCM Cc loi c ph bao gm c ph vi (C. robusta), ch (C. arabica) v mt (C. excelsa) c trng c Lc. 2.2.Cc phng php nghin cu [2] 1) nh lng pectin bng phng php canxi- pectat 2) nh lng cellulose bng phng php axit mnh 3) Xc nh hot tnh pectinase bng phng php so mu c s hin din thuc th Antron 4) Xc nh hot tnh cellulase bng phng php quang ph k c s hin din thuc th DNS (3,5 dinitrosalicylic acid) 5) nh lng ng tng s bng phng php so mu c s hin din thuc th Antron 6) nh lng ng kh bng phng php Schaffer Hartmann 7) nh lng tinh bt bng phng php quang ph k c s hin din thuc th DNS (3,5 dinitrosalicylic acid) 8) nh lng m bng phng php Kjeldahl 3.KT QU V BN LUN 3.1.Thnh phn ha hc c bn trong c ph nhn Ht c ph s dng dng kh bo qun c m thp hn 11W%, c th m ln lt C. robusta, C. arabica v C. excelsa l 7,86, 7,26 v 10,60. Sau khi phn tch thnh phn ha hc trong ht c ph chng ti nhn thy thnh phn pectin v cellulose cao mt cch ng k. Kt qu c trnh by nh trong bng 1 sau y:

Trang 54

TAP CH PHAT TRIEN KH&CN, TAP 11, SO 12 - 2008 Bng 01.Thnh phn ha hc c bn trong c ph nhn
THNH PHN 1. ng tng s ha tan 2. ng kh 3. Pectin 4. Cellulose 5. Tinh bt 6. m
PHN TRM THEO TRNG LNG HT C PH KH (%)

C. robusta 16,03 2,95 54,71 15,29 6.46 1.82

C. arabica 17,09 3,00 52,62 16,80 6.25 1.73

C. excelsa 15,05 4,17 55,14 17,04 6,04 1,96

3.2.Thnh phn mi trng nui cy, nhit , thi gian ti thch cho sinh tng hp pectinase v cellulase Chng ti nhn thy rng c rt c nh hng n vic sinh tng hp pectin v v tru c nh hng n sinh tng hp cellulase. Mi trng nui cy ti u cho vic sinh tng hp pectinase v cellulase l 9% c rt, 15% v tru v nhng thnh phn khc l 75% cm v 1% NH4(SO4)2. Nhit , m v thi gian l nhng yu t quan trng cho vic sinh tng hp pectinase v cellulase. Chng ti tin hnh xc nh cc yu t ny v kt lun rng, iu kin nhit khong 27-300C (nhit phng), m t 56-64%W v thi gian t 40-44 gi l iu kin tt nht cho vic sinh tng hp pectinase v cellulase. 3.3.Cht mang v cc iu kin cho qu trnh nhn ln ca ch phm vi sinh Khi ln men phm vi ln hn, chng ti nhn thy rng ch phm vi sinh cn s dng vi lng ln hn rt nhiu th mi p ng cho nhu cu sn xut cng nghip. V th, chng ti nghin cu v xc nh bt m v bt sn l hai cht ph hp nht cho qu trnh nhn ln ca ch phm vi sinh. Chng ti kt lun rng iu kin tt nht cho qu trnh ny l s dng t l bt m: bt sn l 2:1, thi gian 30 gi v m 56%
2.5
2.277 2.475 2.310 2.376 2.343

Hot tnh enzyme cellulase (UI/g CT)

2.013

2.0

1.5

1.0

0.924

0.5 1 2 3 4 5 Mu (t l bt m : bt sn) 6 7

th 3.1: T l phi trn bt m:bt sn cho sinh tng hp enzyme cellulase

Trang 55

Science & Technology Development, Vol 11, No.12 - 2008

3.5
3.036

3.0 Hot tnh cellulase (UI/g CT) 2.5 2.0

1.551 1.947

2.310 2.145 1.980 1.881

1.5
1.386 1.122 1.023 0.858

1.0 0.5 12 18 24 30 36 42 48 54 60

66

72

Thi gian (gi)

th 3.2: Thi gian ti thch cho qu trnh nhn ging cho sinh tng hp enzyme cellulase
390

370

367.00

Hot tnh enzyme pectinase (UI/g CT)

350

350.00 341.50

330
312.60

310

304.10

306.48

300.70

290 1 2 3 4 5 6 7

Mu (t l bt m : bt sn)

th 3.3: T l phi trn bt m:bt sn cho sinh tng hp enzyme pectinase

390
377.54

370 350
Hot tnh enzyme pectinase (UI/g CT)

330
312.60

310 290 270 250 12

301.38 289.48 283.02

297.30

296.28 288.80 278.94 263.64 256.50

18

24

30

36

42

48

54

60

66

72

Thi gian (gi)

th 3.4: Thi gian ti thch cho qu trnh nhn ging cho sinh tng hp enzyme pectinase

Trang 56

TAP CH PHAT TRIEN KH&CN, TAP 11, SO 12 - 2008 Chng ti kho st iu kin ln men ti u cho vic thu nhn cc cht ha tan. Cc yu t c quan tm bao gm l thi gian ngm ht c ph trong nc, thi gian ngm ht to m ti u, thi gian ln men v t l ch phm. Trong , khi lng cht ha tan c nh ngha l lng cht ha tan trch ra t 10 g c ph qua ln men v t l nc nng s dng l 50ml/10g c ph ln men. Chng ti kt lun rng m c ph cao nht sau 3 gi ngm ht trong nc, tuy nhin lng cht trch ha tan thu c cao nht l sau mt gi ngm ht v thi gian ln men tt nht l 16 gi v t l ch phm / trng lng c ph l: 16/1000.
Bi
50 45 m ht (W%) ) 40 35 30 25 20 15 0.5 1 1.5 2 2.5 3 3.5 4 Thi gian ngm (gi)

Mokka

th 3.5: m theo thi gian ngm ht c ph

Bi
2.3 2.2 Khi lng cht tan (g) 2.1 2.0 1.9 1.8 1.7 0.04

Mokka

0.08

0.12 0.16 T l ch phm (%)

0.20

0.24

th 3.6: Thi gian ti thch cho qu trnh ln men ht c ph

Trang 57

Science & Technology Development, Vol 11, No.12 - 2008

3 Bi S Mokka

Khi lng cht tan (g)

2.5

1.5

1 12 16 20 24 Thi gian ln men (gi) 28 32

th 3.7: T l ch phm ti thch cho qu trnh ln men

Hnh 3.1: C. robusta

Hnh 3.2: C. arabica

Hnh 3.3: C. excelsa (Ngun thu nhn ba loi c ph c Lc )

Trang 58

TAP CH PHAT TRIEN KH&CN, TAP 11, SO 12 - 2008

Hnh 3.4 Mt mu c ph ang ln men

Hnh 3.5 Mt mu c ph sau ln

Hnh 3.6 Mt mu c ph sau khi rang

Hnh 3.7 Mt mu c ph sau khi xay

4.KT LUN Tm li, chng ti kt lun rng biocoffee-1 c th ng dng mt cch hiu qu trong ln men ba loi c ph ph bin Vit Nam, c bit trong vic tng cng trch ly cc cht ha tan trong ht c ph. Nghin cu ny rt hu ch cho cc cng ty sn xut v ch bin lin quan n c ph, c bit cc cng ty ch bin thc ung ha tan.

SYNTHESIZING BIOCOFFEE-1 AND APPLYING IT IN COFFEE FERMENTATION

Le Hong Phu(1), Nguyen Duc Luong(2), Do Dai Nghia(3) (1)International University, VNU-HCM (2) University of Technology, VNU-HCM (3) Nong lam University of HCMc ABSTRACT: In coffee processing today, attracting all the dissolved substances from the coffee bean is a matter that so many manufacturers are interested in. However, the dissolved substrate extraction is recognized by the two major components in the coffee bean: pectine and cellulose as from this research. Pectine contains 52.62-55.14% and cellulose contains 15.29-17.04%. We have successfully created the bio-product named Biocoffee-1, which has high enzymatic activity of pectinase and cellulase. The optimal chemical components of

Trang 59

Science & Technology Development, Vol 11, No.12 - 2008 culture for this bio-product synthesis includes carrot (9 %), rice husk (15%), mash (75%), NH4(SO4)2 (1%) and other factors such as temperature 27-300C (about room temperature), humidity 56-64%W, and time 40-44 hours. On a large scale, when the bio-product is needed for industrial production, it also needs to produce a larger quantity as well. We find that the wheat powder and cassava are the most appropriate for this process. We concluded that the ratio wheat powder: cassava powder is 2:1, humidity 56 %W and bio-synthesis time is optimal at 30 hours. Moreover, we also investigated the optimal fermented conditions for the highest quantity of dissolved substance weight. The dissolved substance weight is defined that the dry quantity of dissolved substance extracted from 10g of fermented coffee and dissolved rate is done by putting 10g of coffee powder in 50ml of hot water. We concluded that coffeebean humidity gets most after 3 hours in water. However; the highest quantity of dissolved substances weight was determined 1 hour after putting coffee in water, and the optimal fermented time was 16 hours and the ratio bio-product weight/ coffee weight is16/1000. Thus, this bio-product can solve the problem as early mentioned and get the high quality of coffee with the fermented method, a new way in coffee processing today. TI LIU THAM KHO [1]. Nguyn c Lng v cc tc gi, Cng ngh enzyme, NXB HQG TP. HCM, (2004) [2]. Nguyn Vn Mi, Thc hnh sinh ha, NXB KH&KT H Ni, (2002) [3]. Gerhartz, Wolfgang, Enzymes in Industry production and applications, VHC publisher, New York, USA, (1990) [4]. Helmut Uglig, Industrial enzymes and their applications, John Wiley & Sons Inc., UK, (1998) [5]. R.J.Clarke, R. Macrae, Coffee, Volume 1: Chemistry, Volume 2: Technology, volume 5:Related beverages, volume 6: Commercial and Technico-legal Aspects, Elsevier applied science publishers London and New York, (1985)

Trang 60