B GIO DC V O TO

TRNG I HC NNG LM TP.H CH MINH

B MN CNG NGH SINH HC
*************

KHA LUN TT NGHIP

BC U NGHIN CU SN XUT BT CACAO

BNG PHNG PHP LN MEN C B SUNG VI SINH VT

Ngnh hc: CNG NGH SINH HC

Nin kha: 2001-2005
Sinh vin thc hin: HONG VN TIN

Thnh ph H Ch Minh
Thng 8/2005

B GIO DC V O TO
TRNG I HC NNG LM TP.H CH MINH
B MN CNG NGH SINH HC
*************

BC U NGHIN CU SN XUT BT CACAO

BNG PHNG PHP LN MEN C B SUNG VI SINH VT

Gio vin hng dn:

Sinh vin thc hin:

PGS. TS. NGUYN C LNG

HONG VN TIN

Thnh ph H Ch Minh
Thng 8/2005

LI CM T

Cm n Ban Gim Hiu trng i hc Nng Lm TP.HCM v tt c cc Ging

Vin trong v ngoi B mn Cng Ngh Sinh Hc tn tnh truyn t kin thc
trong sut bn nm hc ti trng.
Chn thnh bit n thy hng dn tn tnh gip ti ht mnh trong nhng
lc gp kh khn khi lm lun vn.
PGS. TS. NGUYN C LNG
Chn thnh nh n cc thy c v anh ch ti B mn Cng Ngh Sinh Hc
trng i hc Bch Khoa TP.HCM nhit tnh gip ti trong lc ti thc tp ti
trng.
Do hn ch v thi gian cng nh kin thc nn lun vn ny khng th trnh
khi nhng thiu st nht nh, ti rt mong nhn c s gp chn thnh ca qu
thy c v ca cc bn.

TP. H CH MINH, thng 08/2005

HONG VN TIN

iii

TM TT
HONG VN TIN, i hc Nng lm TP. H Ch Minh. Thng 8/2005. BC
U NGHIN CU SN XUT BT CA CAO BNG PHNG PHP LN
MEN C B SUNG VI SINH VT.
Gio vin hng dn:
PGS. TS NGUYN C LNG.
Trong thnh phn ht ca cao ti, hm lng pectin v cellulose chim t l cao
(11% v 16%). Hai thnh phn kh phn gii ny lm cn tr chit cht ha tan ca
bt ca cao. Do , chng ti s dng phng php ln men b sung nm mc Asp.
niger c kh nng sinh pectinase v cellulase cao phn gii hai thnh phn trn gip
lm tng lng cht ha tan ca bt ca cao.
Nhng kt qu t c:
Tuyn chn c ging Asp. niger ly t phng th nghim B mn Cng
ngh sinh hc i hc Bch Khoa TP.HCM c kh nng sinh pectinase (650
vht/g MT) v cellulase (2.3 UI/g MT) thch hp s dng ln men.
Ch phm sinh hc s dng ln men ti u c t l ging Asp. niger : bt m
rang l 1 : 3
Lng ch phm s dng ti u l 1% khi lng ht ln men, m khi ln
men l 60 %, thi gian ln men 96 gi
ha tan ca bt ca cao c ln men c b sung Asp. niger (4.3) cao hn
ha tan ca bt ca cao c ln men khng b sung vi sinh vt (2.1).

iv

MC LC
TRANG

Ba....................................................................................................................................i
Trang ta........................................................................................................................ ii
Li cm t ..................................................................................................................... iii
Tm tt...........................................................................................................................iv
Mc lc ...........................................................................................................................v
Danh sch ch vit tt ................................................................................................ viii
Danh sch cc bng .......................................................................................................ix
Danh sch cc hnh .........................................................................................................x
PHN 1. M U ........................................................................................................1
PHN 2: TNG QUAN TI LIU .............................................................................3
2.1 GII THIU V CY CA CAO........................................................................3
2.1.1
Lch s .......................................................................................................3
2.1.2
Cc c im ca cy cacao ......................................................................5
2.1.2.1 Thc vt hc .................................................................................................... 5
2.1.2.2 c im hnh thi........................................................................................... 5
2.1.2.3 c im sinh thi ........................................................................................... 6
2.1.2.4 Thnh phn ha hc ca tri ca cao................................................................. 7

2.1.3
Tnh hnh trng v sn xut ca cao ti Vit Nam ......................................7
2.2 CNG NGH SN XUT BT CA CAO .....................................................10
2.2.1
Cng ngh ca cao th ..............................................................................10
2.2.1.1 ht ca cao ................................................................................................... 10
2.2.1.2 Cc phng php ht ca cao....................................................................... 12
2.2.1.3 Phi v sy ca cao ......................................................................................... 13

2.2.2

Qui trnh sn xut bt cacao....................................................................14

2.2.2.1
2.2.2.2
2.2.2.3
2.2.2.4
2.2.2.5
2.2.2.6
2.2.2.7
2.2.2.8
2.2.2.9
2.2.2.10

Loi b tp cht ............................................................................................. 14

X l nhit ..................................................................................................... 14
Rang............................................................................................................... 14
Nghin th v phn ly.................................................................................... 16
Kim ho ....................................................................................................... 17
Sy bt kim .................................................................................................. 18
Nghin mnh nhn......................................................................................... 18
p................................................................................................................... 18
Xay bnh du ................................................................................................. 18
Sng .............................................................................................................. 19

2.3

GII THIU V CELLULOSE, PECTIN, CELLULASE, PECTINASE.......19

2.3.1
Gii thiu cellulose .................................................................................19
2.3.2
Gii thiu enzyme cellulase ....................................................................20
2.3.3
Gii thiu v pectin .................................................................................21
2.3.4
Gii thiu enzyme pectinase ...................................................................22
PHN 3: NGUYN LIU V PHNG PHP NGHIN CU .........................25
3.1 THI GIAN V A IM THC HIN TI........................................25
3.2 NGUYN LIU ................................................................................................25
3.2.1
Tri cacao ................................................................................................25
v

3.2.2
Ging vi sinh vt .....................................................................................25
3.2.3
Nguyn liu lm mi trng ...................................................................26
3.3 MI TRNG NUI CY VI SINH VT ....................................................26
3.3.1
Mi trng thch malt.............................................................................26
3.3.2
Mi trng nhn ging............................................................................27
3.3.3
Ch phm sinh hc ..................................................................................28
3.4 DNG C, HA CHT V THIT B..........................................................29
3.4.1
Dng c v thit b ..................................................................................29
3.4.2
Ha cht cho phn tch ............................................................................30
3.5 PHNG PHP NGHIN CU.....................................................................31
3.5.1
Phng php thit k th nghim ............................................................31
3.5.1.1 Tuyn chn ging Asp. niger thch hp nht cho qu trnh ln men ............ 31
3.5.1.2 Nghin cu nh hng ca t l: ging Asp. niger/bt m rang ca ch phm
ln men n ha tan v cng mu ..................................................... 31
3.5.1.3 Nghin cu nh hng ca lng ch phm enzyme s dng ln men n
ha tan v cng mu.......................................................................... 31
3.5.1.4 Nghin cu nh hng ca m ca khi ln men n ha tan v cng
mu ........................................................................................................... 32
3.5.1.5 Nghin cu nh hng ca thi gian ln men n ha tan v cng
mu ................................................................................................................ 32
3.5.1.6 So snh ha tan v cng mu ca bt ca cao t phng php ln men
truyn thng v bt ca cao t phng php ln men c b sung vi sinh vt. 32

3.5.2

Phng php vi sinh vt ..........................................................................32

3.5.2.1 Phng php cy truyn v gi ging ...........................................................32

3.5.2.2 Phng php nhn ging ...............................................................................33

3.5.3

Phng php ha l.................................................................................33

3.5.3.1 Xc nh m.............................................................................................. 33
3.5.3.2 Phng php xc nh ha tan v cng mu ca dung dch ca cao .. 33
3.5.3.3 Phng php xc nh bc sng m dung dch ca cao hp thu mnh nht . 34

3.5.4
3.5.4.1
3.5.4.2

Phng php ha sinh .............................................................................34

Cc phng php xc nh cc thnh phn ch yu ca ht cacao ............... 34
Cc phng php xc nh hot tnh enzyme ca Asp. niger ....................... 39

3.5.5
Qui trnh sn xut bt ca cao trong phng th nghim ............................42
PHN 4: KT QU V THO LUN ....................................................................43
4.1 XC NH CC THNH PHN CH YU CA HT CA CAO..............43
4.2 TUYN CHN GING VI SINH VT THCH HP CHO QU TRNH LN
MEN HT CA CAO.........................................................................................44
4.3 NH HNG CA T L GIA GING VI SINH VT VI BT M
RANG N HA TAN, CNG MU CA BT CA CAO.........45
4.3.1
nh hng n ha tan ......................................................................45
4.3.2
nh hng n cng mu ................................................................46
4.4 NH HNG CA LNG CH PHM SINH HC S DNG LN
MEN N HA TAN, CNG MU CA BT CA CAO ...........47
4.4.1
nh hng n ha tan ......................................................................48
4.4.2
nh hng n cng mu ................................................................49
4.5 NH HNG CA M KHI LN MEN N HA TAN,
CNG MU CA BT CA CAO .........................................................50
4.5.1
nh hng n ha tan ......................................................................50
4.5.2
nh hng n cng mu. ...............................................................51
vi

4.6

NH HNG CA THI GIAN LN MEN N HA TAN, CNG

MU CA BT CA CAO ........................................................................52
4.6.1
nh hng n ha tan ......................................................................52
4.6.2
nh hng n cng mu ................................................................53
4.7 SO SNH HA TAN V CNG MU GIA BT CA CAO
C LN MEN C V KHNG B SUNG VI SINH VT TRONG QU
TRNH LN MEN ............................................................................................54
PHN 5: KT LUN V NGH.........................................................................56
5.1 KT LUN .......................................................................................................56
5.2 KIN NGH.......................................................................................................56
PHN 6. TI LIU THAM KHO ..........................................................................57

vii

Danh sch cc ch vit tt

ACDI: Agricultural Cooperative Development International
VOCA: Volunteers in Overseas Cooperative Assistance
USDA: United States Department of Agriculture
USAID: United States Agency for International Development
PSOM: Programme for Co-operation with Emerging Markets
FAO: Food and Agriculture Organization

viii

Danh sch cc bng

TRANG

Bng 2.1: Sn lng ca cao trn th gii trong cc nm (n v ngn tn) ....................4
Bng 2.2: Cc thnh phn ca ht cacao ti, tnh theo % trng lng ti..................7
Bng 2.3: Cc tnh trng ca cao Vit Nam...................................................................9
Bng 4.1: Cc thnh phn ch yu c trong ht ca cao ti .........................................43
Bng 4.2:So snh hot tnh enzyme cellulase v pectinase do hai ging Asp. niger ly
t trng i hc Khoa Hc T Nhin v i Hc Bch Khoa to ra........44
Bng 4.3: nh hng ca t l gia ging vi sinh vt vi bt m rang n ha tan45
Bng 4.4: nh hng ca t l gia ging vi sinh vt vi bt m rang n cng
mu................................................................................................................46
Bng 4.5: nh hng ca lng ch phm sinh hc n ha tan............................48
Bng 4.6: nh hng ca lng ch phm sinh hc n cng mu......................49
Bng 4.7: nh hng ca m khi ln men n ha tan ....................................50
Bng 4.8: nh hng ca khi ln men n cng mu .........................................51
Bng 4.9: nh hng ca thi gian ln men n ha tan ........................................52
Bng 4.10: nh hng ca thi gian ln men n cng mu ................................53
Bng 4.11: So snh ha tan v cng mu ca hai mu ca cao c v khng b
sung vi sinh vt trong qu trnh ln men.......................................................54

ix

Danh sch cc hnh

TRANG

Hnh 2.1: Cu trc ca cellulose....................................................................................19

Hnh 2.2: S tc ng enzyme cellulase ln cellulose theo Mandels v Reese .......21
Hnh 2.3: Cu trc ca axit polygalacturonic................................................................22
Hnh 2.4: S phn ct ca enzym pectinesterase. ....................................................22
Hnh 2.5: S phn ct ca enzym transeliminase.....................................................23
th 4.1: nh hng ca t l ging/bt m rang n ha tan..............................45
th 4.2: nh hng ca t l ging/bt m rang n cng mu........................47
th 4.3: nh hng ca lng ch phm sinh hc n ha tan ..........................48
th 4.4: nh hng ca lng ch phm n cng mu ..................................49
th 4.5: nh hng ca m khi ln men n ha tan...................................50
th 4.6: nh hng ca m khi ln men n cng mu ............................51
th 4.7: nh hng ca thi gian ln men n ha tan.......................................52
th 4.8: nh hng ca thi gian ln men n cng mu ................................53