Luan Van Sua Chua Bo Sung Tao

NGHIN CU QUY TRNH SN XUT SA CHUA U
NNH TO SPIRULINA
Tc gi
H TH L
Kha lun c trnh p ng yu cu cp bng k s ngnh

Bo Qun Ch Bin Nng Sn Thc Phm v Dinh Dng Ngi
Gio vin hng dn:

Ts. TRNG THANH LONG
Th.S. NGUYN MINH KHANG
7-2009
i
LI CM N
Lng bit n xin chn thnh gi n:
Ban gim hiu v qu thy c trng i Hc Nng Lm tn tnh ging
dy, truyn t nhng kin thc b ch, nhng kinh nghim qu bu cho ti. c bit
l thy Trng Thanh Long tn tnh hng dn, gip ti rt nhiu trong sut thi
gian thc hin lun vn ny.
ng cm n thy Nguyn Minh Khang, trng phng th nghim vi sinh thc
phm trng Cao ng Kinh T Cng Ngh, to iu kin thun li, cho ti nhng
kin b ch trong lun vn ny.
V cng bit n ngi thn, bn b ng vin, gip nhit tnh khi ti thc
hin ti ny.
Do bc u tin hnh lm th nghim ch bin sn phm nn khng trnh
khi nhng thiu st. Knh mong qu thy c v bn b ng gp kin lun vn
c hon chnh hn.
ii
TM TT
ti Nghin cu quy trnh sn xut sa chua u nnh to spirulina
c tin hnh ti Phng th nghim Vi sinh thc phm Trng Cao ng Kinh T
Cng Ngh TP. H Ch Minh, thi gian thc hin t thng 3 n thng 7 nm 2008.
Th nghim c b tr theo kiu hon ton ngu nhin vi 3 ln lp li.
Qua qu trnh th nghim, chng ti thu c mt s kt qu sau
3 Trong mi trng MRS broth, sinh khi vi khun Lactobacillus acidophilus
t cc i sau 15 gi nui cy, sinh khi vi khun Lactobacillus bulgaricus t cc
i sau 18 gi nui cy.
3 Trong mi trng sa, sinh khi vi khun Lactobacillus acidophilus v
Lactobacillus bulgaricus t cc i sau 15 gi nui cy.
3 Thi gian thu men ci vi khun l 6 gi .
3 Cc thng s ca qu trnh ln men
# T l phi ch gia sa u nnh v sa ti UHT l 8:2
# Thi gian ln men 3 gi, 43OC
# Cy 10% men ci vi t l ging cy gia Lactobacillus acidophilus v
Lactobacillus bulgaricus l 6:4
# Hm lng ng saccharose b sung l 8%
# Hm lng to b sung l 6%, gelatin 5%
3 Thi gian bo qun sn phm ti u l 10 ngy 4OC.
iii
MC LC
Trang
Trang ta....................................................................................................................... i
Li cm n..................................................................................................................... ii
Tm tt.......................................................................................................................... iii
Mc lc ......................................................................................................................... iv
Danh sch cc bng ................................................................................................... viii
Danh sch cc hnh ..................................................................................................... ix
Chng 1.M U.....................................................Error! Bookmark not defined.
1.1.t vn ..............................................................................................................1
1.2. Mc tiu ti ......................................................................................................2
1.3.Cng vic thc hin...............................................................................................2
Chng 2.TNG QUAN TI LIU............................................................................3
2.1.Tng quan v u nnh v sa chua u nnh ......................................................3
2.1.1.Lch s pht trin................................................................................................3
2.1.2.Cy u nnh ......................................................................................................3
2.1.3.Ht u nnh.......................................................................................................4
2.1.3.1.Tnh cht c l v cu trc ca ht u nnh .................................................4
2.1.3.2.Thnh phn ha hc ca ht u nnh ............................................................5
2.1.4. Cc sn phm t u nnh v li ch ca sa chua u nnh .........................10
2.2.Tng quan v to spirulina ..................................................................................11
2.2.1.Lch s ..............................................................................................................11
2.2.2.Phn loi ...........................................................................................................12
2.2.3.c im sinh hc ca Spirulina .....................................................................12
2.2.3.1.c im cu to t bo ca to spirulina .....................................................12
2.2.3.2. Sinh sn ca to Spirulina: ...........................................................................13
2.2.3.3.Thnh phn ha hc ca to spirulina ...........................................................14
2.2.3.4.Tnh hnh sn xut, tiu th v ng dng ca to spirulina...........................17
2.3. Tng quan v vi sinh vt ....................................................................................19
2.3.1. Probiotic ..........................................................................................................19
2.3.2. Vi khun ln men sa chua .............................................................................19
iv
2.3.2.1. Lactobacillus acidophilus ............................................................................20

2.3.2.2. Lactobacillus bulgaricus .............................................................................20
2.4. Tng quan v sa v cht n nh ......................................................................21
2.4.1. Sa ..................................................................................................................21
2.4.2. Cht n nh ...................................................................................................21
2.5. Cc yu t nh hng n qu trnh ln men ....................................................21
Chng 3.VT LIU V PHNG PHP NGHIN CU .................................22
2.1. Thi gian v a im th nghim.......................................................................22
2.2. Nguyn liu ........................................................................................................22
2.3. Dng c, thit b v ha cht s dng ................................................................22
2.4.Quy trnh ch bin sa chua u nnh to Spirulina...........................................23
2.4.1.Quy trnh...........................................................................................................23
2.4.2.Gii thch quy trnh...........................................................................................24
2.5. Phng php th nghim.....................................................................................26
2.5.1.Xy dng ng cong tng trng ca Lactobacillus acidophilus,
Lactobacillus bulgaricus trn mi trng MRS. ......................................................27
Lactobacillus bulgaricus trn mi trng dch sa (sa u nnh v sa ti vi t
l 1:1).........................................................................................................................28
2.5.3.Lm men ci. ....................................................................................................29
2.5.4. Kho st qu trnh ln men..............................................................................30
2.5.4.1. Kho st thi gian ln men. ..........................................................................30
2.5.4.2. Kho st t l phi ch gia sa u nnh v sa ti tit trng khng
ng.........................................................................................................................30
2.5.4.3. Kho st hm lng ng saccharose b sung. .........................................30
2.5.5. Kho st t l phi ch gia dch sa ( sa ti UHT + sa u nnh) vi dch
to spirulina...............................................................................................................31
2.5.6. Kho st thi gian bo qun sn phm............................................................31
2.6. Phng Php Phn Tch Cc Ch Tiu...............................................................32
2.6.1. Mt t bo vi sinh vt: ................................................................................32
2.6.2. Xc nh pH ca sn phm:.............................................................................32
v
2.6.3. Phng php chun acid.............................................................................32

2.6.4. Phng php nh gi cm quan sn phm ....................................................32
2.6.5. Phng php b tr th nghim v x l kt qu .............................................32
Chng 4.KT QU V THO LUN...................................................................33
4.1. ng cong tng trng ca Lactobacillus acidophilus, Lactobacillus
bulgaricus trn mi trng MRS broth....................................................................33
4.1.1. Hnh thi khun lc trn mi trng MRS agar ..............................................33
4.1.2. ng cong tng trng ca Lactobacillus acidophilus, Lactobacillus
bulgaricus trn mi trng MRS broth....................................................................33
bulgaricus trn mi trng dch sa (sa u nnh v sa ti vi t l 1:1) ........36
4.3. Kho st qu trnh lm men ci. .........................................................................38
4.4. Kho st qu trnh ln men.................................................................................39
4.4.1. Kho st thi gian ln men ..............................................................................40
4.4.2. Kho st t l phi ch gia sa u nnh v sa ti tit trng khng ng
...................................................................................................................................41
4.4.3. Kho st hm lng ng saccharose b sung .............................................42
4.4.4. Kho st t l ging cy ...................................................................................43
4.5. Kho st hm lng to b sung ........................................................................44
4.6. Kho st thi gian bo qun sn phm...............................................................45
4.7. Kt qu phn tch cc ch tiu vi sinh v ha hc ..............................................49
4.7.1. Kt qu phn tch ch tiu vi sinh ....................................................................49
4.7.2. Kt qu phn tch ch tiu ha hc ..................................................................49
Chng 5.KT LUN V NGH .......................................................................50
5.1. Kt lun...............................................................................................................50
5.2. ngh ...............................................................................................................50
TI LIU THAM KHO...........................................................................................51
PH LC .....................................................................................................................53
vi
DANH SCH CC BNG

Bng 2.1: Thnh phn ha hc trung bnh ca cc hp phn trong ht u nnh
Bng 2.2: Thnh phn ha hc cc cht trong ht u nnh
Bng 2.3: Thnh phn cc acid amin khng thay th trong u nnh v mt s thc
phm quan trng
Bng 2.4: Thnh phn ha hc ca Spirulina
Bng 2.5: Thnh phn vitamin trong Spirulina
Bng 2.6: Thnh phn khong trong Spirulina
Bng 2.7: Thnh phn acid amin trong Spirulina
Bng 2.8: Cc cht mu trong Spirulina
Bng 2.9: Tnh hnh sn xut Spirulina trn th gii
Bng 4.1: Kt qu tng trng ca vi khun L. acidophilus theo thi gian nui cy
trong mi trng MRS
Bng 4.2: Kt qu tng trng ca vi khun L. acidophilus theo thi gian nui cy
trong mi trng dch sa
Bng 4.3: Kt qu tng trng ca vi khun L. bulgaricus theo thi gian nui cy
Bng 4.4. Kt qu pH men vi khun L. acidophilus v L. bulgaricus qua thi gian kho
st
Bng 4.5: M t cm quan men vi khun qua thi gian kho st
Bng 4.6: Kt qu pH ca dch sa theo thi gian ln men
Hnh 4.8: Kt qu pH ca dch sa theo thi gian ln men
Bng 4.6: M t cm quan dch sa theo thi gian ln men
Bng 4.7: im trung bnh kt qu nh gi cm quan sn phm khi thay i t l phi
ch gia u nnh v sa ti
Bng 4.8: im trung bnh kt qu nh gi cm quan sn phm khi thay i hm
lng ng b sung
Bng 4.9: im trung bnh kt qu nh gi cm quan sn phm khi thay i t l
ging cy
Bng 4.10: im trung bnh kt qu nh gi cm quan sn phm khi thay i hm
lng to b sung
vii
Bng 4.11: Kt qu kho pH, acid v s lng vi sinh vt theo thi gian bo qun
Bng 4.12: im nh gi th hiu trung bnh ca ngi th cho cc c tnh ca sn
phm
Bng 4.13: Kt qu kim tra ch tiu vi sinh sn phm
Bng 4.14: Kt qu kim tra cc ch tiu ha hc
viii
DANH SCH CC HNH

Hnh 1. 1: nh hng ca pH n kh nng ha tan ca protein u nnh
Hnh 2.2: S vng i ca Spirulina
Hnh 4.1: L. acidophilus
Hnh 4.2: L. bulgaricus
Hnh 4.3: ng cong sinh trng ca L. acidophilus trong mi trng MRS broth.
Hnh 4.4: ng cong sinh trng ca L. bulgaricus trong mi trng MRS broth
Hnh 4.5: ng cong sinh trng ca L. acidophilus trong mi trng dch sa
Hnh 4.6: ng cong sinh trng ca L. bulgaricus trong mi trng dch sa
Hnh 4.7: pH men vi khun L. acidophilus v L. bulgaricus qua thi gian kho st
Hnh 4.9: b sung 6% to
Hnh 4.12: Kt qu PH v s lng vi khun lactic theo thi gian bo qun sn phm
Hnh 4.13: Kt qu acid ca sn phm theo thi gian bo qun
Hnh 4.14: im nh gi th hiu ca ngi th cho hai sn phm
ix
Chng 1
M U
1.1.t vn
Ngy xa, khi t nc cn ngho v lc hu chng ta thng nghe cu ni n
no mc . Nhng ngy nay, t nc ang trn pht trin th cu ni n no mc
c thay th bng cu n ngon mc p. Tuy nhin,nhu cu con ngi khng
ch dng n ngon m h cn i hi cao hn l n ngon b dng v a dng
thc phm.
Sa u nnh l mt loi sa lm t u nnh, theo cc nh nghin cu vic
kin tr dng sa u nnh hng ngy s lm gim c cholesterol trong mu, h
dc huyt p v phng x cng ng mch ngi ln tui. (Tm Diu,
1998).Ngoi lng ln protein, trong sa u nnh cn cha n 8 loi acid amin an
trng i vi c th con ngi ( leucine, isleucine, lysine, methionine, phenyl alanine,
threonine, tryptophane, valine). Acid bo khng no trong u nnh cn c tc dng
phng chng li cc mm ung th nh: Bowman-Bird Inhibitor (BBI), protease
inhibitor, phytate, phytosterol, isoplavones, cht phenolic acid,(Tm Diu, 1998).
Sa l mt loi thc phm giu dinh dng, cung cp gn nh y cc
vitamin v khong cht cho c th. Nh nhng tc dng c ch m sa v cc sn
phm t sa rt c mi ngi a chung. Th trng sa v cc sn phm t sa
kh phong ph v chng loi, p ng cho mi ngi mi tui.
Sa chua, sn phm ln men t sa, ngi ta da vo quy lut u tranh sinh
tn t nhin tng cng sc kho cho con ngi, cc vi khun lactic c a vo
rut s cnh tranh vi vi khun gy thi v ln t cc vi khun ny. Theo cc nghin
cu th sa chua rt c li cho c th, n c tc dng chng lo ha, gim cholesterol
trong mu, hn ch ri lon ng rut, lm gim tc dng ph do s dng thuc
khng sinh, lm tng interferon, gim vim lot d dy, ngn nga long xng
To l nhm cc vi sinh vt c kh nng quang t dng, chng tham gia tng
hp ngun nng lng sng s cp t nh sng mt tri. To Spirulina c nhiu u
im vt tri c bit v gi tr dinh dng v c cha nhiu hp cht c gi tr trong

phng v iu tr mt s bnh.
Chnh v nhng l do trn, mc ch ca ti Nghin cu quy trnh sn xut
sa chua u nnh to spirulina nhm to to ra sn phm mi a dng v cm quan
v c tc dng b dng.
1.2. Mc tiu ti
To ra sn phm sa chua va c gi tr dinh dng va c gi tr cm quan.
1.3.Cng vic thc hin
3 Xy dng ng cong tng trng ca Lactobacillus acidophilus,
Lactobacillus bulgaricus trn mi trng MRS.
3Xy dng ng cong tng trng ca Lactobacillus acidophilus,
Lactobacillus bulgaricus trn mi trng dch sa.
3Kho st cc yu t ca qu trnh ln men: kho st thi gian ln men, t l
phi ch gia sa u nnh v sa ti, kho st hm lng ng saccharose b
sung, kho st t l ging cy.
3Kho st hm lng to b sung.
3Kho st thi gian bo qun sn phm.
Chng 2
TNG QUAN TI LIU
2.1.Tng quan v u nnh v sa chua u nnh
2.1.1.Lch s pht trin
u nnh c xem l mt trong nhng cy trng c nht ca nhn loi. Nhiu
nghin cu chng minh rng u nnh c ngun gc t Trung Quc. N c trng
trn nhiu loi t khc nhau v c phm vi kh hu rng khp, tri di t vng nhit
i Brazil n hn o Hokkaido y tuyt nm pha bc Nht Bn. Vi s pht
trin ca khoa hc k thut, con ngi tm thy vai tr ca u nnh trong nhiu
lnh vc, c bit l trong lnh vc thc phm. V vy, u nnh c lan rng ngy
cng nhiu trn khp th gii.[ 23]
Yogurt ting Th Nh K c ngha l s ln men, thc phm c lng acid nh
c ln men bi vi khun lactic nh Lactobacillus bulgaricus, Streptococcus
thermophilus, Lactobacillus acidophilus. Vo cui nhng nm 1970, M, sn phm
t sa u nnh c gi theo nhiu cch khc nhau: soymilk yogurt, soy yogurt,
soyogurt, soygurt. Nm 1912, ti Pari, Li Yu Ying v Grand Voinnet ch bin
sa u nnh ln men. Nm 1974, nghin cu ca Wang v Konishi tch ging L.
acidophilus thch hp ch bin soy yogurt. Nm 1977, Sterm, Hesseltine, Wang v
Konishi chn ging L. acidophilus tn dng Raffinones, Stachyose, hn hp ng
trong sa u v dng L. acidophilus lm soy yogurt c v ngon (pH = 4,2), gim y
hi. Nm 1981 ti Nht Bn ln u tin sn xut soy yogurt s lng ln. [ 24]
2.1.2.Cy u nnh
Cy u nnh c tn khoa hc l Glycine max (L) Merrill, thuc h
Leguminosae, chng Papilionoidae, l cy bi nh, cao trung bnh di 1m, c lng
ton thn. L c 3 cht hnh bu dc. Chm lng mc nch l, bng c mu trng
hoc tm. Tri c nhiu lng vng, di 3 4 cm, rng 0,8 cm, mi tri c t 3 5 ht.
Cy u nnh l cy ngn ngy, pht trin tt nht vng nhit i, a sng, a nhit,
chu hn.[ 1], [ 21]
iu kin cy u nnh pht trin tt :
9 pH ca t trng: 6,0 6,5.
9 Nhit : 25 30oC.
9 Lng ma: 500 700 mm.
9 t trng c nhiu mn hu c vi m trung bnh.
9 Thi k trng: cui ma xun, u ma h.
2.1.3.Ht u nnh
2.1.3.1.Tnh cht c l v cu trc ca ht u nnh [ 7]
Tnh cht c l nh hng rt ln n quy trnh ch bin cng nh cht lng
sn phm. Mt s tnh cht ch yu l:
3 Hnh dng: ht u nnh c hnh dng trn, bu dc, trn di, trn dtty
tng ging.
3 ln ca ht cng rt khc nhau ty c im ca ging v k thut trng
trt, thng t 220 300g /1000 ht.
3 Kch thc ht: ty thuc vo ging v iu kin canh tc.
3 V ht nhn phn ln mu vng, mt s c mu vng m, xanh, nu hoc
en. V ch chim khong 8% khi lng ca ht
Cu trc ht u nnh gm c 3 b phn chnh: v, tr di l mm, l mm.
3V: l lp ngoi cng ca ht u nnh. V ht u nnh c nhiu loi mu
sc v l yu t cho vic xc nh ging u nnh. V ht c tc dng bo v phi
mm chng li nm v vi khun.
3Tr di l mm: l r mm, phn sinh trng ca ht khi ht ny mm.
3L mm: gm hai l mm tch tr dinh dng cho ht. N chim phn ln
khi lng ca ht, cha hu ht cc protein, lipid ca ht, c mu xanh trc khi ht
chn, v chuyn thnh mu vng khi ht chn.
Bng 2.1: Thnh phn ha hc trung bnh ca cc hp phn trong ht u nnh
(% so vi khi lng cht kh ca ht) [ 9]
Protein
( Nx 6,25)
Lipid
Glucid
( C x)
Khong
Tng
L mm
43
23
29
90
8,8
86
4,3
Tr di l mm
41
11
43
4,4
Ht nguyn
40
20
35
4,9
100
2.1.3.2.Thnh phn ha hc ca ht u nnh

So vi cc ngun thc phm chnh hin nay, u nnh thc s l thnh phn
giu dinh dng. u nnh va c ngun protein cao nht trong cc loi u, ht; giu
cht bo, carbohydrate, cc cht x, vitamins, cht khong va l dc liu qu cho
vic ngn nga v cha tr cc bnh mn tnh.
Bng 2.2: Thnh phn ha hc cc cht trong ht u nnh
(so vi khi lng cht kh) [ 21]
Thnh phn
PROTEIN
AMINOACID
n v
Hm lng
Hm lng trung
bnh
%
g/100g ht
30 50
40
1,49 1,87
2,45 3,49
3,87 4,98
6,10 8,72
1,88 2,02
0,56 0,66
1,88 2,61
1,81 2,32
1,69
2,90
4,48
7,26
1,69
0,60
2,02
2,07
0,89 1,08
1,46 2,12
2,71 3,20
2,35 2,86
0,49 0,66
1,70 - 2,08
1,33 1,79
0,47 0,54
1,12 1,62
1,52 2,24
1,04
1,76
3,03
2,58
0,54
1,95
1,58
0,49
1,43
1,83
12 30
20
4 - 23
3 - 30
11
4
Khng cn thit
Alanine
Arginine
Aspartic acid
Glutamic acid
Glycine
Cysteine
Proline
Serine
Cn thit
Histidine
Isoleucine
Leucine
Lysine
Methionine
Phenylalanine
Threonine
Tryptophan
Tyrosine
Valine
LIPID
ACID BO
%
%so vi lipid tng
Palmitic acid
Stearic acid
5
Oleic acid
Linoleic acid
Linolenic acid
25 - 86
25 - 60
1 - 15
25
53
7
26 - 38
34
2,5 - 8,2
0,1 - 0,9
1,4 - 4,1
5,5
0,9
3,5
4,61 - 5,94
5,0
g/g
g/g
g/g
6,26 - 6,85
0,92 - 1,19
10,9 - 28,4
ISOFLAVONE
0,2
SAPONINS
0,1 - 0,3
PHYTATE
1,0 - 1,5
1,1
TRYPSIN
INHIBITORS
mg/g
16,7 - 27,2
22,3
PHYTOSTEROLS
mg/g
0,3 - 0,6
CARBOHYDRATE
Sucrose
Raffinose
Stachyose
TRO
VITAMIN
Thiamine
Riboflavin
Vitamin E
Protein
Trong thnh phn ha hc ca u nnh, thnh phn protein chim mt t l rt
ln. Thnh phn acid amin trong protein ca u nnh, ngoi methionin v tryptophan
ra cn cc acid amin khc, c s lng kh cao tng ng lng acid amin c trong
tht.
Bng 2.3: Thnh phn cc acid amin khng thay th trong u nnh v mt s
thc phm quan trng (g/100g protein)
u nnh
Trng
Tht b
Sa b
Go
Gi tr c ngh
bi FAO - OMS
Leusine
7,84
8,32
8,00
10,24
8,26
4,8
Isoleusine
4,48
5,60
5,12
5,60
3,84
6,4
Lysine
6,40
6,24
2,12
8,16
3,68
4,2
Phenylalanine
4,96
5,12
4,48
5,44
4,80
2,8
Loi acid amin
Threonine
3,84
5,12
4,64
4,96
3,36
2,8
Tryptophane
1,28
1,76
1,21
1,44
1,28
1,4
Valine
4,80
7,52
5,28
7,36
5,76
4,2
Methionine
1,28
3,20
2,72
2,88
2,08
2,2
Da vo chc nng sinh hc thc vt, protein trong ht u nnh c chia

lm 2 loi: protein chuyn ha (metabolic protein) v protein d tr. Protein chuyn
ha gm enzyme v cu trc. Protein d tr cng vi du d tr c tng hp trong
qu trnh pht trin ca ht u. Protein u nnh ch yu l protein d tr. Hai loi
protein d tr chnh trong u nnh l glycinin (11S) v beta-conglycinin (7S) chim
khong 65 80% protein tng trong ht.[ 15], [ 21]
Nu phn loi theo tnh tan trong nc, protein u nnh cha 85 95%
globuline, mt lng nh albumine, mt lng khng ng k prolamine v glutelin.
Kh nng ha tan trong nc ca protein u nnh nh hng nhiu bi pH. Khong
80% protein trong ht u nnh c th c trch ly pH trung tnh hoc kim. pH
ng in ca protein u nnh l 4,2 4,6. [ 21]
% Protein ha tan
Hnh 1. 1: nh hng ca pH n kh nng ha tan ca protein u nnh

Cc cht phi dinh dng trong thnh phn protein u nnh: trypsin inhibitors
v hemagglutinins (lectins) [ 21]
3 Trypsin inhibitors l hp cht khng bn nhit cao. Cht ny c kh
nng kt hp vi trypsin lm trypsin khng c hp thu. Thiu trypsin s lm c th
7
chm pht trin v nh hng nhiu n cc chc nng khc ca c th. u nnh
cha 2 loi trypsin inhibitors: Bowman Brik inhibitor v Kunitz inhibitor
3 Lectin (hemagglutinins) l protein c kh nng kt dnh cc t bo hng cu.
N c xem nh l cht c c tnh cao, tuy nhin n cng d dng b v hot di
tc dng ca nhit cao.
Cc tnh cht chc nng ca protein u nnh: hp th v gi nc, to gel, to
kt cu, to nht, to nh, to bt.
3Kh nng hp th v gi nc: Kh nng hp th v gi nc ca protein da
trn tng tc gia protein protein v protein nc. Nng protein tng, kh
nng hp th nc tng. pH thay i th s tch in ca protein cng thay i, pH
ng in s ht nc l thp nht v tng tc gia protein protein rt cht ch,
pH cao s hp th v gi nc cng tng. Nhit tng, kh nng hp th nc gim
v lm gim lin kt hydro.
3Kh nng to gel: Khi protein b bin tnh, cc cu trc bc cao b ph hy,
cc mch polypeptide dui ra, tin li gn nhau, tip xc v to nn mng li khng
gian tng i cht v pha phn tn cha trong n.
Cc yu t to gel
+ S dng nhit: Khi dch sa protein u nnh c nng cao c un nng
pH trung tnh, th s to gel.
+ S dng pH ng in: pH ng in protein c kh nng to gel.
+ S dng cc mui ca ion kim loi ha tr hai: Do lin kt gia Ca2+ v nhm
carboxyl.
3Kh nng to kt cu: Protein u nnh c kh nng to si tt. Khi protein b
phn ly, chui polypeptide dui mch, v vy khi cho qua khun c, s nh hng
c cc phn t protein v cc si hnh thnh.
3Kh nng to nht: Cc yu t nh hng n nht ca dung dch gm:
pH, nhit , ion Ca2+ v nng protein.
3Kh nng to nh: Trong phn t protein u nnh c hai u a nc v a
bo, nn c kh nng nh ha vi cht bo.
3Kh nng to bt: Kh nng to bt l do s khuch tn protein n b mt
tip xc gia pha kh v nc.
Lipid
Cht bo chim khong 20% trng lng cht kh ca ht u nnh. N nm
ch yu trong phn t dip ca ht. Cht bo cha khong 6,415,1% acid bo no
(acid stearic, acid archidic) v 8093,6% acid bo khng no (acid linoleic, acid
linolenic, acid oleic trong acid linolenic chim t l tng i cao). Cht bo khng
no c li cho sc khe hn l cht bo no. Tuy nhin cc acid bo khng no d b oxy
ha v gim cht lng.
Carbohydrate
Hydratcarbon chim khong 34% ht u nnh. Phn hydratcarbon c th chia
ra lm 2 loi: loi tan trong nc v khng tan trong nc. Loi tan c trong nc
ch chim khong 10% ton b hydratcarbon.
Trong cc nhm ng ca u nnh ta cn lu hai loi ng raffinose v
stachyose. C th con ngi thiu enzyme galactosidase thy phn stachyose v
rafinose. Hai loi ng ny thng di chuyn t rut non sang rut gi l ni c th
ln men ng bi vi khun ng rut. Qu trnh ln men to ra cc kh gm CO2, H2
v CH4 lm cho c th ngi cm thy y hi. S loi b hay gim hin tng ny
i vi cc thc phm t u nnh cn phi c quan tm. Trong qu trnh ln men
sa u nnh c th lm gim hm lng raffinose v stachyose. [ 11]
Vitamin v khong cht
Mc d u nnh khng c xem nh l ngun rt giu mt loi vitamin no
nhng n em li mt ngun dinh dng ton din. Cc vitamin c trong u nnh
nh l thiamine, riboflavin, niacin, pantothenic acid, biotin, folic acid, inositol,
choline, vitamin A v E, tr vitamin C v vitamin D.
Thnh phn khong chim khong 5% trng lng kh ca ht u nnh. Cc
cht khong ch yu c trong u nnh l K, Na, Ca, Mg, S, v P. Hm lng cht
khong c th thay i nhiu do loi u cng nh iu kin trng trt khc nhau.[ 21]
Enzyme
V mt cng ngh, enzyme quan trng ca u nnh l lipoxygenase, c bit
n l lipoxydase. Enzyme ny xc tc cho phn ng oxy ho acid bo khng bo ho
bi O2, gy mi cho u nnh. Kh mi u nnh da trn nguyn tc v hot enzyme
lipoxygenase bng cch s dng ring r hay kt hp cc phng php nhit (sy,
chn), phng php ha hc (NaHCO3).
Enzyme urease c tm thy vi hm lng ln u nnh sng. N s phn
hy ure thnh ammoniac, l mt hp cht c i vi c th ngi. Enzyme ny s
chng li s hp th cc cht m qua mng rut, do khng nn n u nnh sng.

Hn na, enzyme urease d dng b v hot bi nhit.[ 6]
Isoflavone
Isoflavones l hp cht phenolic c xem nh l phytoestrogens, hay
estrogens thc vt v chng c cng thc ha hc, chc nng v nh hng n c th
con ngi nh estrogen.
Probiotic vi khun lactic khi pht trin c th chuyn ha glycoside isoflavone
thnh dng aglycone m khng cn phi b sung thm cht dinh dng no. Trong sa
u nnh cha ln men giu glycoside, dng aglycone thp hn. Ngc li, sa u
nnh sau ln men li c hm lng aglycone cao hn so vi glycoside do cc vi khun
lactic c kh nng tit ra enzyme - glucosidase chuyn ha glycoside thnh
aglycone. Isoflavone aglycone hp thu nhanh hn v nhiu hn so vi dng glucoside
trong c th ngi, gip ngn chn cc bnh mn tnh nh ung th v cc bnh v tim
mch. [ 12]
Li ch ca isoflavone u nnh [ 25]
3Ngn cn v cha tr ung th v, ung th tuyn tin lit v ung th rut.
3Gim kh nng hot ng ca hormone gii tnh hormone ny kch thch
kh nng pht trin ung bu c nam v n.
3Gim nguy c bnh tim mch, chng long xng v tng cng kh nng t
min dch.
2.1.4. Cc sn phm t u nnh v li ch ca sa chua u nnh
Cc sn phm t u nnh [ 23]
3Sa u nnh: u nnh ti c ngm vi nc, sau ly dch lc un 20
pht nhit 900C s c sa u nnh n gin. Sa u nnh rt giu protein,
isoflavones v vitamin B, c th l thc ung thay th cho nhng ngi d ng vi sa
b.
3Tofu: c to thnh bi s ng t sa u nnh bng cht lm ng. Tofu
c ba dng: dng rn rt giu protein, cht bo v calcium, dng nu, nng, chin;
dng mm v dng mn dng trn lm thc n, nh Nht, ngi ta dng dng
mm v mn lm nc xt u nnh.
3Yoghurt: l sn phm ca s ln men sa u nnh bng vi khun.
10
3Tempeh: l dng bnh c to thnh bi s nu u v trn vi vi khun, lo

ha (ageing) mt hay hai ngy. Tempeh rt giu protein v khong cht, isoflavones
(100 g tempeh c 53 mg isoflavones).
3Miso: l sn phm ca s kt hp u nnh vi mui, mc v sau trong
thng g 1 3 nm.
3Shoyu: l cht lng mu nu sm, c lm t u nnh qua qu trnh ln
men.
Li ch ca sa chua u nnh [ 18]
3 Vi khun ln men sa u nnh c th s dng c cc ng raffinose,
stachyose c trong sa u nnh lm gim cc triu chng y hi do cc ng ny
gy ra.
3vi khun trong sa ln men cn sng gip tng cng h min dch v to ra
cc enzyme thy phn protein thnh cc polypeptid, thy phn tinh bt thnh cc
maltodextrin lm sn phm tr nn d tiu ha hn.
3 Probiotic trong sa ln men cn lm cn bng h vi sinh vt trong ng
rut, tng cng cc vi khun c li, gim cc vi khun c hi gy bnh cho ngi.
Nhiu nghin cu chng minh rng mt s vi khun lactic c kh nng chuyn ha
isoflavone glucoside (c nhiu trong u nnh) thnh cc isoflavone aglycone l cht
c hot tnh sinh hc, c tc dng lm gim nguy c gy bnh ung th, tim mch, gim
cc triu chng ph n mn kinh nh bc ha, long xng,
2.2.Tng quan v to spirulina
2.2.1.Lch s
Spirulina l tn gi do nh to hc ngi c Deurben t vo nm 1827 da
trn hnh thi c trng nht l dng si xon c vi khong 5-7 vng u nhau khng
phn nhnh. V cng vo nm 1827, Turpin ln u tin phn lp c Spirulina t
ngun nc t nhin.
Nm 1973, T chc Nng lng Quc t (FAO) v T chc Y t Th gii
(WHO) chnh thc cng nhn Spirulina l ngun dinh dng v dc liu qu, c
bit trong chng suy dinh dng v chng lo ha.
11
Nm 1977, Vin sinh vt hc l ni tin phong trong vic nui trng Spirulina
Vit Nam theo m hnh ngoi tri, khng mi che, c sc kh CO2 ti x nghip nc
sui Vnh Ho (Bnh Thun).
2.2.2.Phn loi[ 2], [ 5], [7]
To (algae) l mt nhm vi sinh vt, nhng chng khc vi vi khun v nm
men ch chng c dip lc v c kh nng tng hp c cc cht hu c t cc
cht v c di tc dng ca nh sng mt tri.
To chia lm 9 ngnh: to lam (Cyanophyta); to lc (Chorophyta); to silic
(Diatomea); to vng nh (Chysophyta); to gip (Pynophyta); to mt
(Euglenophyta); to roi lch (Hererocontac); to (Rhodophyta); to nu.
To Spirulina thuc:
Ngnh: Cyanophyta
Lp: Cyanophyceae
B: Oscillatoriales
H: Oscillatoniaceae ( Nostocales)
Ging: Spirulina
2.2.3.c im sinh hc ca Spirulina [ 2], [ 5]
To Spirulina c dng xon l xo khong 5 - 7 vng u nhau khng phn
nhnh. ng knh xon khong 35 - 50 m, bc xon 60 m, chiu di thay i c
th t 0,25 mm.
To l trung gian gia vi khun v to nhn thc. Ngi ta cho rng to
Spirulina ging vi vi khun hn, do to Spirulina cn c tn l vi khun lam.
To c kh nng vn chuyn theo hnh thc trt xung quanh trc ca chng.
Vn tc vn chuyn ca chng c th t 5 micron/ giy.
2.2.3.1.c im cu to t bo ca to spirulina
L to lam a bo dng si, gm nhiu hnh tr xp khng phn nhnh. mi t
bo ca si c chiu rng 5 m, di 2 mm.
Khng c lc lp m ch cha thylacoid phn b u trong t bo.
Khng c khng bo. Khng c nhn in hnh, vng nhn khng r, trong
c cha DNA (Hedeskog v Hifsten A.1980).
Thnh t bo c cu trc nhiu lp cha mucopolymer, pectin v cc loi
polysaccharide khc.Mng t bo nm st ngay di thnh t bo v ni vi mng
quang hp thylacoid ti mt vi im.
12
B my quang hp ca Spirulina:
3Phycobilisome: cha phycobiliprotein v protein lin kt c gn vo b mt
ngoi ca thylacoid. Phycobilisome c khi lng khong 7 triu dalton v c th tch
nguyn vn nghin cu. i vi phycobilisome c c phycoerythin v phycocianin
th lp ngoi cng l phycoerythin, tip theo l phycocianin v phn trong cng l
allophycocyanin.
3Phycobilisome hot ng nh mt anten thu nhn nng lng mt tri
chuyn vo PS II. Con ng truyn nng lng bt u t phycoerythin sang
phycocyanin v cui cng n allophycocyanin trc khi t ti PS II.
3C khong 50% nng lng nh sng mt tri, Spirulina nhn c nh
phycobilisome.
3Cc sc t quang hp gm chlorophylla, carotenoid, phycocianin v thng
c carotenoid-glycoside nh myxoxanthophyll, oscillaxanthin.
Spirulina c cha 3 nhm sc t chnh:
3Chlorophyll hp ph nh sng lam v .
3Carotenoid hp th nh sng lam v lc.
3Phycobilin hp th nh sng lc, vng v da cam.
2.2.3.2. Sinh sn ca to Spirulina: [ 3], [ 5], [ 13]
Spirulina c phng thc sinh sn v tnh, t mt c th m trng thnh (gi
l trichome), t phn chia thnh nhiu mnh, mi mnh gm mt s vng xon (2-4 t
bo, gi l hormogonia). to thnh cc hormogonia, si Spirulina s hnh thnh cc
t bo chuyn bit cho s sinh sn (gi l on Necridia). Cc necridia hnh thnh cc
a lm 2 mt v to ra hormogonia bi s chia ct ti v tr cc a. Khi pht trin,
dn dn phn u hormogonia b tiu gim v tr nn trn nhng vch t bo vn c
chiu dy khng i. Cc hormogonia pht trin, trng thnh v chu k sinh sn lp
li m bo vng i ca Spirulina
Thng thng Spirulina sinh sn bng cch gy ra tng khc. Trong trng hp
gp iu kin khng thun li, Spirulina cng c kh nng to bo t ging nh vi
khun. Chu k pht trin ca Spirulina rt ngn. Chu k ny thng din ra trong 24
gi nh ca to Chlorella
13
Hnh 2.2: S vng i ca Spirulina [Ciferri, 1985]

Spirulina l sinh vt phiu sinh sng t do trong nc, pht trin tt trong kh
hu m, mi trng kim v c bit l nhng ni c nhiu nng nh Kona, Hawai
2.2.3.3.Thnh phn ha hc ca to spirulina
Spirulina cha hm lng protein rt cao v cha y cc vitamin.
Spirulina c gi tr dinh dng cao v cha hm lng protein cao v cc cht
c hot tnh sinh hc khc. Gi tr protein trung bnh ca Spirulina l 65%, cao hn so
vi nhiu loi thc phm. V d, hm lng protein ca c v tht l 15-20%, nc
tng l 35%, sa c c l 35%, trng l 12% v ca ng cc l 8-14% (R. Herehson,
Earth Food Spirulina, Konore Press,1977).
Bng 2.4: Thnh phn ha hc ca Spirulina [ 5]
STT
Thnh phn
S lng (% cht kh)
Protein tng s
60 70
Glucid
13 16
Lipid
78
Acid nucleic
4,29
Dip lc
0,76
Carotene
0,23
Tro
45
Spirulina l ngun giu vitamin B12 nht. Nu hng ngy chng ta s dng 1g
Spirulina th s p ng nhu cu vitamin B12 hng ngy. Ngoi ra, Spirulina cn
cha cc vitamin khc nh A, B1, B2, B6, E v H (Fox, 1986). Spirulina cung cp 21%
thiamin v riboflavin so vi nhu cu hng ngy. Provitamin A (-caroten) (chim
14
0,1% cht kh) cao hn 20 ln so vi trong carrot. Thnh phn cc vitamin ca

Spirulina c lit k trong Bng 2.2.
Bng 2.5: Thnh phn vitamin trong Spirulina [ 17]
Vitamin
Trn 10g
23000 IU
Nhu cu hng
ngy cho php
5000
% so vi nhu cu
hng ngy cho php
460
Vitamin A ( -carotene)
Vitamin B1 (Thiamine)
0,31 g
1,5
21
Vitamine B2 (Riboflavin)
0,35 g
1,7
21
Vitamin B3 (Niacin)
1,46 g
20
Vitamin B6 (Pyridoxine)
80 g
2,0
Vitamin B12
32 g
6,0
533
Citamine E (-tocoferol)
1 IU
30
Folacin
1 g
400
0,04
Panthothenic acid
10 g
10
Biotin
0,50 g
Inositol
6,40 g
(Cyanocobalamine)
Spirulina giu st v calcium, h tr tt cho mu, cho xng v rng. Lng

calcium ca Spirulina cao hn trong sa (Fox, 1986). Lng st trong Spirulina cao
hn 12 ln so vi trong cc loi thc phm khc. Ngoi ra, Spirulina giu magnesium,
potassium. Nhng khong a lng bao gm sodium, calcium, magnesium, potassium,
chlorine, sulfur v phosphorous. Cc khong vi lng gm iodine, zinc, copper,
selenium, molybdenum, fluoride, manganese, boron, nickel v cobalt. Lng Ka v Ca
chim lng ln nht trong cc khong a lng (160 g v 100 g/10g Spirulina),
trong cc khong vi lng th Mn chim hm lng cao nht (500 g/10g Spirulina).
Bng 2.6: Thnh phn khong trong Spirulina [ 17]
Khong
Trn 10g
Nhu cu hng ngy
% so vi nhu cu hng ngy
Calcium
100 g
1000 g
10
Iron
15 g
18 g
83
Zinc
300 g
15 g
15
Phosphorous
90 g
1000 g
Magnesium
40 g
400 g
10
Copper
120 g
2 g
Sodium
60 g
2 - 5 g
Potassium
160 g
6 g
Manganese
500 g
3 g
17
Selenium
2 g
100 g
Spirulina cha 18 trong s 20 loi amino acid c bit (Fox, 1986). Mt s

amino acid c hm lng cao trong Spirulina nh glutamic acid (14,6%); aspartic acid
(9,8%); leucine (8,7%); aniline (7,6%)
Bng 2.7: Thnh phn acid amin trong Spirulina [ 5]
Acid amin
Hm lng
Cc Acid amin
Hm lng
thit yu
trong 10g
khc
trong 10g
Phenylalanine
280 g
4,5 %
Glycine
320 g
5,2 %
Threonine
320 g
5,2 %
Histidine
100 g
1,6 %
Tryptophan
90 g
1,5 %
Proline
270 g
4,3 %
Valine
400 g
6,5 %
Serine
320 g
5,2 %
Isoleucine
350 g
5,6 %
Tyrosine
300 g
4,8 %
Leucine
540 g
8,7 %
Alanine
470 g
7,6 %
Lysine
290 g
4,7 %
Arginine
430 g
6,9 %
Methionine
140 g
2,3 %
Aspartic Acid
610 g
9,8 %
Cystine
60 g
1,0 %
Glutamic Acid
910 g
14,6 %
%/tng
%/tng
Spirulina c mu xanh lam-lc l do Spirulina cha nhiu sc t vi hm lng

cao nh chlorophyll, phycocyanin, beta-caroten, xanthophyll (Estrada v cng s,
2001)
16
Bng 2.8: Cc cht mu trong Spirulina [ 5

Cht mu
Mu sc
Hm lng trong 10g
% Spirulina
Phycocyanin
Xanh da tri
1400 g
14 %
Chlorophyll
Xanh l cy
100 g
1,0 %
Carotenoids
Mu vng cam
47 g
0,47%
2.2.3.4.Tnh hnh sn xut, tiu th v ng dng ca to spirulina

Spirulina c coi l ngun thc n ca con ngi trong nhiu th k. Spirulina
sp. l ging c bit n nhiu nht v gi tr dinh dng ca n. Sinh khi to c
s dng ch yu nh ngun cung cp protein cho ngi v gia sc. Nht Bn, ngi
ta sn xut sinh khi to c nhn hiu Linablue A, M c Phycobiliprotein.
Bng 2.9: Tnh hnh sn xut Spirulina trn th gii [ 2]
Cng ty
a im
Din tch
Sn lng
Gi thnh
(ha)
(tn kh)
(USD/kg)
Sosa-Texcoco
Mehico
12
300
Earthrise
Hoa K
5,5
340
Cyanotech
Hoa K
170
Siam Algae
Thi Lan
3,8
480
Blue Continent
i Loan
480
Nippon Spirulina
Nht Bn
1,5
40
Bnh Thun
Vit Nam
0,5
Nhiu cng ty
Trung Quc
200
2798
Nhiu cng ty
12,2
260
Nhiu cng ty
Cu Ba
40
17
5 - 18
ng dng ca to spirulina:
3Spirulina c s dng nh mt ngun thc phm t hng ngn nm nay
cung cp dinh dng v ci thin sc khe cho con ngi v Spirulina cha nhiu cht
dinh dng nh protein, acid amin, acid bo cn thit, vitamin, cht khong [ 20]
3 Nm 1970-1980, nhiu nghin cu s dng Spirulina phng v tr bnh
suy dinh dng tr em c tin hnh Mehico, Php, Roumani, Nht Bn, Hoa
K, Peru [ 3]
3Bt kh Spirulina c dng nh mt thc n cao cp giu protein, acid amin
v nhiu vi lng b dng khc. Nu xt v cht lng protein, ch cn lng 30 40g Spirulina/ngy/ngi l nhu cu acid amin. Spirulina dng ti tt hn
Spirulina kh do cn nguyn vn cc dng cht, cc enzyme ang hot ng gip ch
cho c th nh superoxydedismutase, protease, lipase,v vic s dng Spirulina
dng ti cn mang li ch kinh t v khng phi sy kh, bo qun, thch hp vi m
hnh nui Spirulina h gia nh hoc lin kt vi x nghip, sn xut vi trng hc,
nh tr, khoa dinh dng ca bnh vin [ 3]
3Theo Gloria v cng s (2004), khi b sung thm sinh khi to S. platensis
dng kh vi hm lng (6 mg/ml) vo sa s kch thch s pht trin ca Lactococcus
lactis ln 27%, do b sung Spirulina thc y qu trnh ln men lactic [ 16]
3Ch phm Enalac (bt dinh dng b sung 5% Spirulina) l mt ch phm do
Trung tm Dinh dng tr em Tp.HCM sn xut. Enalac cha 16,5% protein; 7%
lipid; 74% glucide, vitaminvi s phi ch ca 5% bt Spirulina. Khi s dng, ch
phm c pha long trong nc m (50-60oC) c s dng n trc tip hoc
nui qua sonde. Kt qu nghin cu cho thy ch phm c tc dng tt cho bnh nhn
suy dinh dng mi la tui [ 3]
3 Mt s nghin cu cho thy Spirulina c tc dng tt trong iu tr bnh bo
ph, bnh tiu ng, bnh thiu mu, bnh lot d dy t trng, vim ty, bnh c
thy tinh th v suy gim th lc, bnh rng tc. Trong iu tr bnh bo ph, ch phm
Spirulina c ch nh ung vo trc cc ba n khong 1 gi v Spirulina c cha
nhng cht vi lng tc dng iu ha ni tit t (hormon), gip duy tr lng glucose
trong mu mc cn thit, gip c th vn khe mnh v khng c cm gic thm n [
3]
18
2.3. Tng quan v vi sinh vt

2.3.1. Probiotic [ 14]
Probiotics l thut ng ny xut hin trong nhiu thp k qua v ang tr
nn quen thuc vi con ngi. Probiotic l cc vi khun sng khi c cung cp vi
liu lng thch hp s em li li ch sc khe cho vt ch (theo nh ngha ca FAO
v WHO 2001).
Vai tr ca probiotic
3Tng thnh bo v min dch, mt s c kh nng kch thch c min dch
c hiu v khng c hiu.
3Ngn cn s pht trin ca cc tc nhn gy bnh trong ng rut nh:
Staphylococcus, Salmonella, Yersinia, Clostridia.
3C kh nng xm chim ng rut, bm vo mng nhy rut.
3C kh nng chu c acid d dy, chu c mui mt.
3Phng v cha mt s bnh ng tiu ha: tiu chy, to bn, ung lot d
dy
3Hn ch s hnh thnh cc khi u. Enzym c tit ra bi cc vi khun c hi
s gp phn to ra cc cht gy ung th. Probiotic c kh nng lm gim ti mc ti
thiu hot ng ca cc enzym ny v gim nguy c ung th rut.
3Mt s c th khng s dng c lactose do khng c enzym lactase hoc
lactase hot ng khng tt, iu ny dn ti s kh tiu khi dng cc sn phm sa.
Probiotic c kh nng sn xut ra enzym lactase gip tiu ho lactose d dng.
3Vic s dng nhiu thuc khng sinh l nguyn nhn gy ra cc bnh tiu
chy, nhim Candida do h vi sinh vt c ch trong ng rut b tiu dit. Probiotic
gip ti thit lp h vi sinh vt c ch nhanh chng sau khi dng khng sinh.
2.3.2. Vi khun ln men sa chua
Vi khun ln men sa chua l vi khun lactic in hnh, thng l cc loi
Lactobacillus bulgaricus , Lac. Acidophilus, Streptococcus lactis, streptococcus
cremosus Nhng vi sinh vt ny khi ln men sa to ra sn phm sa chua c li
cho sc khe ca con ngi, l nhng Probiotics.
19
2.3.2.1. Lactobacillus acidophilus [ 22]

L. acidophilus l vi khun Gram dng, c dng hnh que, di, mnh c kch
thc t 0,6-0,9 x 1,5-6 m, thng xp theo cp hoc to thnh chui ngn. Khun
lc trn mi trng thch c kch thc 2-5 mm, li, c, u v khng sinh cht to
mu. c im chung ca h vi khun lactic l khng sinh bo t.
L. acidophilus l loi t c kh nng di ng, khng phn gii protein, phn ng
catalase v chuyn ha nitrat thnh nitrit m tnh. L. acidophilus ln men ng
lactose, v cc loi ng glucose, saccharose, maltose, mannose.
L. acidophilus l loi vi khun vi hiu kh.
Nhit ti u l 370C.
pH ti u cho s tng trng dao ng trong khong 5,5 6. Nhu cu dinh
dng ca L. acidophilus phn nh bn cht rt kh nui cy ca vi khun ny. Mi
trng nui cy chun thng phi rt giu acit amin v vitamin nh peptone, trypton,
dch chit nm men, dch chit tht b, ngoi ra cn cha sorbitol, monooleate (tween
80), sodium acetate v mui magne kch thch s tng trng. Mi trng nui cy
hay c s dng l MRS (De Man, Rogosa, Sharpe) lng hay agar.
2.3.2.2. Lactobacillus bulgaricus
Lactobacillus bulgaricus l trc khun gram dng, hnh que, di, mnh, thng
xp thnh i hay chui di t 5- 20 m, khng c kh nng di ng, c th to ra
2,7% acid lactic.
Khun lc ca Lactobacillus bulgaricus phng, hi vng, ng knh 2- 3mm.
Khun lc cng gi th gia cng tr nn sm mu. Trn mi trng thch- huyt
thanh sa cho khun lc hnh trn dng si bng ri.
Lactobacillus bulgaricus khng dch ha gelatin, lm ng t sa 37OC.
Chng ln men c ng glucose, lactose v galactose. Hm lng acid do vi khun
ny to ra trong mi trng sa rt cao (3-3,5%, tng ng 30-45OT). N thuc loi k
kh khng bt buc. Nhit ti u cho s pht trin ca chng l 40-48OC , pH thch
hp 5,4. Khi ln men chng khng c kh nng s dng ng maltose v to mt
lng nh acid bay hi v aldehyde, to cho sn phm c mi v c trng, thng
dng sn xut sa chua ng mn.
20
2.4. Tng quan v sa v cht n nh

2.4.1. Sa [ 8]
Sa l mt loi thc phm rt giu cht dinh dng. V vy n l loi thc phm
qu, c bit i vi ngi gi v tr em.
Trong sa c khong 90% l nc v c cc cht dinh dng nh protein, cht
bo, ng lactoza, cc vitamin nh A, D, E, B1, B6, B12, v cc cht khong. Trong
sa cha un cn c mt s enzym gip cho vic tiu ha c d dng.
Hin nay sn phm sa ch yu l sa b vi cc loi a dng nh sa ti, sa
hp, sa hp pastuer, sa c, kem, b, phomat, sa chua, sa bt.
2.4.2. Cht n nh [ 4]
Cc cht n nh thng dng: gelatin, pectin, tinh bt, agar-agar. Chng lin kt
vi nc, tng nht, ngn s tch nh thanh. Yoghurt bnh thng khng cn thm
cht n nh. Yoghurt tri cy c th thm cht n nh mc 0,1-0,5%.
2.5. Cc yu t nh hng n qu trnh ln men [ 11]
Trong cng nghip sn xut sa chua, cc giai on x l sn phm sau qu trnh
ln men thng n gin. Sn phm cui cng bao gm c sinh khi vi sinh vt, cc
sn phm trao i ngoi bo do vi sinh vt tit ra v c cht st. Thnh phn v hm
lng tng cht s gp phn hnh thnh nn hng v cho sn phm. Do , sn
xut c sa chua vi cht lng mong mun, chng ta cn phi:
- Chn ging vi khun thch hp.
- S dng mi trng ln men vi thnh phn c cht y theo t l ti u.
- Ln men trong iu kin thch hp nh: lng ging cy, nhit v thi gian
ln men. Ngoi ra, ta cn phi quan tm n nhng vn khc nh vic cung cp
oxy, s khuy trn
21
Chng 3
VT LIU V PHNG PHP NGHIN CU
2.1. Thi gian v a im th nghim
Cc th nghim c thc hin ti phng th nghim vi sinh thuc khoa Cng
Ngh Sinh Hc trng Cao ng Kinh T Cng Ngh s 547/M8 Nguyn Tri
Phng, Phng 14, Qun 10, Thnh ph H Ch Minh.
Thi gian thc hin ti t thng 03 n thng 07 nm 2009.
2.2. Nguyn liu
Ht u nnh c chn t u nnh Phng Lm. Nguyn liu c chn mua
siu th Nguyn Vn C, ng V Vn Ngn, qun Th c, Tp.HCM.
Sa c chn mua l sa tit trng khng ng ca Vinamilk, mua siu th
Nguyn Vn C, ng V Vn Ngn, qun Th c, Tp.HCM
To spirulina c chn l bt to thng mi c ngun gc Nht Bn, c
ng gi ti cng ty Chu i Dng ( 79/3X Phan Ty H, Phng 7, Qun Ph
Nhun, Tp.HCM), loi hp 50g.
ng c chn mua l ng ct trng tinh luyn, mua siu th Nguyn
Vn C, ng V Vn Ngn, qun Th c, Tp.HCM.
Ging vi khun Lactobacillus acidophilus, Lactobacillus bulgaricus ca vin
Pasteur Tp. H Ch Minh.
2.3. Dng c, thit b v ha cht s dng
Dng c
Thit b
Ha cht
3Nhit k
3Khc x k ATAGO 0
3Mi trng MRS agar (
3Ni nu
32oBx
ph lc)
3Bp un
3T sy
3Mi trng MRS broth
3Vi lc
3Cn in t
(Ph lc)
3Ray
3Cn ng h
3Cn
3Que cy tri
3My xay sinh t
3Paraffin ca cng ty AR
22
3a Petri
3My ng ha
Trung Quc
3Bnh tam gic c chia
3T gi lnh ca cng ty
3Phenoltalein 1%
vch
Toshiba Nht Bn
3Dung dch NaOH 0,1N
3a thy tinh
3T m( c th chnh nhit
3Micropipette, u tip
thnh t sy) ca cng
3Buret
ty Membert- c.
3ng nghim
3Ni autoclave SA 252F

ca cng ty Sturdy i
Loan
3T cy ca cng ty Huy
Hong Vit Nam
3My o pH pH211 ca
cng ty Hanna Italia
2.4.Quy trnh ch bin sa chua u nnh to Spirulina

2.4.1.Quy trnh
u nnh
La chn
Bt to
X l
Dch sa
X l
Phi trn
Dch to
ng
Sa UHT
khng ng
ng ha
23
Thanh trng
Vi khun
lactic
Nhn ging
Cy ging
Rt h
Sn phm
Bo qun
2.4.2.Gii thch quy trnh

La chn nnh: u c chn thng l u gi, rut vng, v mng, da
lng. Loi b nhng ht h hng ( su, mt, mi...), ht km phm cht ( ht non, ht
khng bnh thng v mu sc v hnh dng...) do nhng ht ny s lm cho sn phm
c mi v rt km.
X l: X l u nnh to ra dch sa bao gm cc cng on sau
3Ngm: trong giai oi ngm, ht s ht nc v trng ln, khi cc phn
t nc c tnh lng cc s tc ng ln cc cu phn a nc trong ht.
Kt thc giai on ngm l thi im m ht t 55 -60% l tt nht(Nguyn
c Lng ,1996). Thi gian ngm u c chn l 4 gi.
Giai on ngm cn gip cho cng on loi b v trong iu kin khng c
my tch v mt cch d dng hn.
3Bc v: bc v bng cch i th cng trong nc. Bc v nhm mc ch
lm cho qu trnh chn hiu qu hn v sa u nnh thu dc trng hn.
3Chn: chn nng ht u trong dung dch Natribicarbonat nhm loi b yu
t LOX ( lipoxigenase, l nguyn nhn gy ra mi kh chu), h tr qu trnh trng
n, ly trch protein, c ch cht khng dinh dng. Chn dung dch u trong dung
24
dch NaHCO3 0,2% nhit 80OC trong vng 5 pht. T l dung dch/ u l 5/1.
Sau khi chn, u c ra li bng nc nng 80OC.
3Xay v lc: xay t u l qu trnh ph v t bo gii phng protein, lipit,
gluxit v.v...ng thi dng nc ha tan cc cht v chuyn chng sang dng huyng
ph. Sau lc huyn ph gm dung dch keo v cht rn khng tan thu dch sa.
Xay u trong nc lnh trong thi gian 1 pht 30 giy vi t l nc/u khi xay l
6/1. Sau khi xay u c lc qua vi lc.
3X l nhit: c tin hnh ngay sau khi lc hn ch nh hng ca s tip
xc oxi khng kh, m, vi sinh vt.Gia nhit sa u nnh c cc yu cu sau cn t:
tiu dit vi sinh vt, dit enzyme, dit cht khng dinh dng, c t, kh mi tanh
u nnh sng. X l nhit 95OC trong vng 15 pht.
X l bt to: Bt to c cn v pha vo nc theo t l 3g to / 50 ml nc
ct m ( khong 40- 50OC), lc ly dung dch to b sung vo dch sa.
Phi trn: Phi trn sa u nnh, sa ti, dch to, ng saccharose theo
nhng t l nht nh.
Trong qu trnh th nghim, t l phi trn gia cc thnh phn nh sau: dch
sa l s phi trn gia sa u nnh v sa ti vi t l 1:1, lng dch to b sung
l 6% (v/v), lng ng saccharose b sung l 6% (w/v), ngoi ra b sung thm 5%
gelatin dng bt ( w/v).
ng ha: nhm to nn mt hn hp ng nht, ng ha tc 4 trong
vng 5 pht.
Thanh trng:thanh trng 80OC trong vng 15 pht.
Nhn ging vi khun: Trc khi nhn ging ta phi lm thun v gi ging vi
khun.
3Lm thun ging l cng vic tch ri mt loi t bo t qun th sinh vt
c phn lp, cy chng ln b mt mi trng thch dinh dng chn lc (hoc trn
thch vo trc khi ng ri ra a Petri). Sau khi cc iu kin thch hp, cc
t bo s phn chia nhiu ln to thnh nhng qun th ring bit gi l khun lc. Mt
t t bo trong khun lc c cy vo ng mi trng gi ging v cc iu kin
nh trn chng pht trin thnh mt qun th mi.Qun th t bo trong ng
nghim ny c gi l mt chng hoc ng ging.
25
Lm thun v gi ging c tin hnh nh sau: Vi khun trong ng ging mua

t vin Pastuer c cy ria trn mi trng MRS agar, 37oC trong t m 2 ngy
cho n khi thun chng, tin hnh gi ging trong ng thch nghing hoc thch su.
3Nhn ging:
Ging vi khun
c lm thun
Nhn ging cp 1
(MRS lng)
37oC, 15 gi
Cho ging vi khun lm

quen mi trng sa
Nhn ging cp 2
( 1 sa u nnh + 1 sa
UHT khng ng)
10% ging, 43oC, 6 gi
Men ci
Cy ging: Cho vo dch sa men ci vi t l 10% ( T l gia men L.
acidophilus v L. Bugaricus l 1:1)
: Sa c 43OC trong thi gian khong 4- 8 gi.
Bo qun: Sn phm c bo qun nhit 4OC.
2.5. Phng php th nghim
Trong nghin cu ny, chng ti tin hnh nghin cu quy trnh ch bin sa
chua u nnh to Sprirulina vi tm lc phng php nghin cu theo s sau:
26
Xy dng ng cong sinh

trng ca vi khun trn mi
trng MRS
Xy dng ng cong sinh

trng ca vi khun trong dch
sa ( sa ti UHT v sa u
nnh).
Kho st thi gian ln men
Lm men ci
Kho st t l phi ch gia sa
ti UHT v sa u nnh
Kho st qu trnh ln men
Kho st hm lng ng
saccharose b sung
Kho st hm lng to b sung
Kho st t l ging cy
Kho st thi gian bo qun sn

phm
Xc nh cc ch tiu vi sinh v
ha hc

Lactobacillus bulgaricus trn mi trng MRS.
a/ Mc ch
Xc nh thi gian nui cy tt nht thu c s lng vi khun nhiu nht.
b/ B tr th nghim
27
Thi gian
(gi)
10
15
20
Vi khun
Lactobacillus bulgaricus
Lactobacillusacidophilus
c/Ch tiu theo di

S lng vi khun sau cc khong thi gian nui trong mi trng MRS lng.
d/ Phng php thc hin
- Ging vi khun trong mi trng MRS agar cy chuyn qua mi trng MRS
lng, 37OC trong cc khong thi gian b tr nh trn.
-S lng vi khun c xc nh bng phng php a.( Ph lc 1.1)
Lactobacillus bulgaricus trn mi trng dch sa (sa u nnh v sa ti vi
t l 1:1)
a/ Mc ch
Kho st s lng vi khun cn sng trong mi trng sa sau cc khong thi
gian nui cy, t chn ra khong thi gian nui cy tt nht thu c s lng vi
khun cao nht.
b/ B tr th nghim
Thi gian
(gi)
10
15
20
Vi khun
c/Ch tiu theo di

S lng vi khun sau cc khong thi gian nui trong mi trng sa u
nnh v sa ti vi t l 1:1.
28
d/ Phng php thc hin

- Ging vi khun trong mi trng MRS agar cy chuyn qua mi trng MRS
lng, 37OC trong thi gian ti u.( ng dng kt qu th nghim 2.5.1).
-Ly vi khun trong mi trng MRS lng cho vo mi trng sa ti khng
ng, 37OC trong 4h.
-Vi khun t mi trng sa ti c cy vo mi trng sa ti v sa tit
trng phi trn vi t l 1:1, 37OC qua cc khong thi gian v xc nh s lng
vi khun.
-S lng vi khun c xc nh bng phng php a.
2.5.3.Lm men ci.
a/ Mc ch
To ra h men t yu cu v s lng vi khun v gi tr cm quan.
b/ B tr th nghim
Xc nh gi tr pH v nh gi cm quan sn phm qua bng sau:
Thi gian
(gi)
Men ci vi
khun
c/ Ch tiu theo di
-gi tr pH
-nh gi cm quan sn phm v mu sc, mi v, cu trc.
d/ Phng php thc hin
-Vi khun t trong mi trng MRS lng c cy chuyn qua mi trng sa
ti UHT v 43OC trong 4 gi.
-Tip tc cy chuyn vi khun t mi trng sa ti sang dch sa (sa u
nnh v sa ti UHT vi t l 1:1), 43OC trong cc khong thi gian b tr nh
trn.
-Kho st cm quan sn phm theo thi gian.
29
2.5.4. Kho st qu trnh ln men.

2.5.4.1. Kho st thi gian ln men.
Chun b sa u nnh ging mc 2.4.2 v ng dng kt qu cc th nghim
trn
Theo di pH, c im sn phm theo thi gian v kt hp vi ng cong sinh
trng ca vi khun trong sa u nnh, t chn thi gian ln men thch hp sao
cho sn phm m bo lng vi khun cn thit n ng vai tr nh l probiotic
2.5.4.2. Kho st t l phi ch gia sa u nnh v sa ti tit trng khng
ng.
a/ Mc ch
Phi ch gia sa u nnh v sa ti nhm to ra dch sa tng hm lng
protein v lm cho sn phm sa chua to ra c mi thm hn, v bo hn.
b/ B tr th nghim
Th nghim mt yu t gm 3 nghim thc
Yu t th nghim l t l phi ch gia sa u nnh v sa ti UHT nh sau :
4:6, 5:5, 6:4.
S ln lp li: 3 ln
c/ Ch tiu theo di
-pH ca sn phm.
-Kt qu nh gi cm quan v cu trc, mi v v sn phm theo phng php
so hng.
d/ Phng php thc hin
Mi th nghim thc hin theo quy trnh nu. Nhng t l phi ch gia sa
u nnh v sa ti UHT thay i theo cc thng s nu nh trn.
2.5.4.3. Kho st hm lng ng saccharose b sung.
trn
Ln lt b sung ng saccharose vo sa theo cc t l kho st khc nhau
6%, 8%, 10%(w/v)
Theo di pH v nh gi cm quan sn phm theo phng php so hng. T
chn hm lng ng thch hp b sung vo sa.
30
2.5.4.4.Kho st t l ging cy.

trn
Thay i t l ging cy t l gia Lactobacillus acidophilus v Lactobacillus
bulgaricus thay i nh sau 6:4, 5:5, 4:6 (v/v).
Theo di pH v nh gi cm quan sn phm theo phng php so hng. T
chn t l ging cy thch hp b sung vo sa ln men.
2.5.5. Kho st t l phi ch gia dch sa ( sa ti UHT + sa u nnh) vi
dch to spirulina.
a/ Mc ch
Kho st nh hng ca hm lng to phi ch hng ln n mu, mi v v
sn phm.
b/ B tr th nghim
Th nghim mt yu t gm 3 nghim thc
Yu t th nghim l t l phi ch gia dch to spirulina v dch sa (sa u
nnh + sa ti UHT) vi cc t l nh sau:6%, 8%, 10%(v/v)
S ln lp li: 3 ln
c/ Ch tiu theo di.
-Kt qu nh gi cm quan v cu trc, mu, mi v v sn phm.
d/ Phng php thc hin
Mi th nghim thc hin theo quy trnh nu. Nhng t l phi ch gia dch
to v dch sa (sa u nnh + sa ti UHT) thay i theo cc thng s nu nh
trn. ng dng kt qu cc th nghim trn.
2.5.6. Kho st thi gian bo qun sn phm
Sn phm c bo qun nhit 4OC
Theo di PH, acid v s lng vi khun lactic trong sn phm theo cc ngy
bo qun chn ra c thi gian bo qun ti u cho sn phm ( sn phm va t
v gi tr cm quan va t c gi tr dinh dng v probiotic).
Sau thi gian bo qun ti u, tin hnh nh gi cm quan sn phm theo
phng php cho im th hiu nhm xc nh mc chp nhn sn phm ca ngi
tiu dng.
31
2.6. Phng Php Phn Tch Cc Ch Tiu

2.6.1. Mt t bo vi sinh vt:
c xc nh theo cng thc sau:
N
C (CFU/g) =
n1xV1xf1 ++ n1xV1xf1
Trong :
C (CFU/g) l s t bo vi khun trong 1g mu

N ( CFU) l tng s khun lc m c trn cc a
ni l s a cy nng th i
Vi (ml) l th tch mu cy trong mi a
fi l pha long th i
2.6.2. Xc nh pH ca sn phm:
o bng my o pH Hanna 210.
2.6.3. Phng php chun acid
Cho 10ml sa vo bnh tam gic 100ml, sau cho vo khong 3 git
phenoltalein 1%. Dng NaOH 0,1N chun ( dung dch xut hin mu hng phn
trong 10 giy).
% acid= n. 0,09
n: s ml NaOH 0,1N
2.6.4. Phng php nh gi cm quan sn phm
3Phng php so hng. ( ph lc 1.3.1 )
3Phng php cho im. (ph lc 1.3.2 )
2.6.5. Phng php b tr th nghim v x l kt qu
3B tr th nghim 1 yu t.
3Kt qu thu c x l bng phn mm statgraphic7.0.
32
Chng 4
KT QU V THO LUN
bulgaricus trn mi trng MRS broth
4.1.1. Hnh thi khun lc trn mi trng MRS agar
Hnh 4.1: L. acidophilus
Hnh 4.2: L. bulgaricus
Khun lc L. acidophilus trn mi trng MRS agar c dng hnh trn, kch
thc 2 - 5 mm, li,mu trng c, u v khng sinh cht to mu.
Khun lc L. bulgaricus trn mi trng MRS agar phng, hi vng, ng
knh 2 - 3mm.
4.1.2. ng cong tng trng ca Lactobacillus acidophilus,
Lactobacillus
bulgaricus trn mi trng MRS broth

Chng ti tin hnh dng ng cong sinh trng ca vi khun Lactobacillus
acidophilus, Lactobacillus bulgaricus trn mi trng MRS broth, nhm mc ch
nghin cu nh tnh vi sinh vt theo thi gian v xc nh thi im mt t bo t
cc i t tip tc tng sinh trong mi trng sa u nnh.
Vi khun c nui cy trong mi trng MRS nhit 370C, pH = 5,7.
Ging c cy trc tip t ng gi ging thch nghing.
33
Bng 4.1: Kt qu tng trng ca vi khun L. acidophilus theo thi gian nui
cy trong mi trng MRS
Thi gian
(gi)
0
3
6
9
12
15
18
21
24
27
30
33
36
Mt vi khun
L. acidophilus
Log (CFU/ml)
1,89
2,33
4,81
6,08
7,3
8,33
8,37
8,34
8,3
8,28
8,22
7,8
7,31
Mt vi khun
L. bulgaricus
Log (CFU/ml)
5
5,3
6
6,95
7,08
7,15
8,2
8,08
8,18
8
7,7
6,96
6
9
8
Log( CFU/ml)
7
6
5
4
3
2
1
0
0
10
20
30
40
Thi gian( gi)
Hnh 4.3: ng cong sinh trng ca L. acidophilus

trong mi trng MRS broth.
34
Nhn xt:
T ng cong sinh trng ca vi khun L. acidophilus chng ti nhn thy:
pha tim pht ko di trong khong 2 - 3 gi u do s lng t bo vi khun tng ln
khng ng k. di pha tim pht tng i ngn chng t cc t bo vi khun cn
tr, nhanh chng thch nghi vi mi trng MRS broth. T 3 - 15 gi, vi khun sinh
trng v pht trin theo ly tha, vi khun sinh sn mnh, sinh khi tng nhanh, y
l pha cp s ca vi khun. S lng t bo vi khun t cc i 15 gi, l thi im
bt u pha cn bng. Pha cn bng ko di t 18 - 30 gi, trong pha ny s sinh sn
vn tip tc din ra mt s t bo trong mi trng nui cy, tng s t bo mi
c sinh ra bng vi tng s t bo cht b mt i, cc cht dinh dng trong mi
trng bt u cn kit, cng vi s tch ly nhiu acid lactic c th gy c ch s pht
trin ca chnh vi khun. Sau 30 gi mt t bo gim dn, bt u pha suy vong, do
hm lng cht dinh dng trong mi trng cn kit, s cnh tranh cc cht dinh
dng ca vi khun din ra, cng vi sn phm ca qu trnh trao i cht c ch
s pht trin ca vi khun nn s lng t bo sinh ra t hn s lng t bo mt i.
9
8
Log( CFU/ml)
7
6
5
4
3
2
1
0
0
10
20
30
40
Thi gian( gi)
Hnh 4.4: ng cong sinh trng ca L. bulgaricus

trong mi trng MRS broth
Nhn xt:
T ng cong sinh trng ca vi khun L. acidophilus chng ti nhn thy:
pha tim pht ca L. bulgaricus cng ko di trong khong 2 - 3 gi u. Trong 3 - 18
35
gi tip theo l pha cp s. V s lng t bo t cc i 18 gi. Pha cn bng din

ra trong 18 - 30 gi. Sau 30 gi l pha suy vong ca vi khun.
Kt lun:
Chng ti thu nhn sinh khi vi khun L. acidophilus sau 15 gi nui cy v thu
nhn sinh khi L. bulgaricus sau 18 gi nui cy trong mi trng MRS broth.
bulgaricus trn mi trng dch sa (sa u nnh v sa ti vi t l 1:1)
Chng ti tin hnh xy dng ng cong tng trng ca Lactobacillus
acidophilus, Lactobacillus bulgaricus trong mi trng dch sa nhm kho st s
lng vi sinh vt trong mi trng dch sa trong cc khong trong thi gian nui cy
ng dng vo lm men ci cho qu trnh sn xut sa chua.
Bng 4.2: Kt qu tng trng ca vi khun L. acidophilus theo thi gian nui
cy trong mi trng dch sa
Mt vi khun L. acidophilus
Log (CFU/mL)
7,86
8,92
10,69
11.36
10.26
9.08
12
10
pH
4
6
3
4
2
1
0
0
10
15
20
Log( CFU/ ml)
Thi gian
(gi)
0
5
10
15
20
25
pH
6,35
5,9
5,1
4,2
4
3,8
pH
Log s lng
( CFU/ ml)
25
Thi gian( gi)
Hnh 4.5: ng cong sinh trng ca L. acidophilus trong mi

trng dch sa
36
Bng 4.3: Kt qu tng trng ca vi khun L. bulgaricus theo thi gian nui cy
Mt vi khun L. acidophilus
Log (CFU/mL)
7,56
8,85
10
10,6
9,9
9,48
12
10
pH
4
6
3
4
Log( CFU/ml)
Thi gian
(gi)
0
5
10
15
20
25
pH
6,36
6,1
5,15
4,4
4,1
3,9
pH
Log s lng
( CFU/ ml)
1
0
0
0
10
15
20
25
Thi gian( gi)
Hnh 4.6: ng cong sinh trng ca L. bulgaricus trong mi trng

dch sa
Nhn xt:
T ng cong sinh trng ca L. acidophilus v L.bulgaricus chng ti nhn
thy: s lng t bo vi khun ngay ti thi im cy vo mi trng dch sa > 107
CFU/ml. S lng t bo tng nhanh trong 15 gi tip theo kt hp vi s gim mnh
gi tr pH do acid lactic ca vi khun sinh ra trong qu trnh trao i cht. S lng t
bo vi khun t cc i 15 gi. Sau 15 gi s lng t bo gim do hm lng cc
37
cht dinh dng trong mi trng gim v lng acid lactic trong mi trng nhiu c
ch s pht trin ca vi khun.
Kt lun:
Trong mi trng dch sa, vi khun vn sinh trng v pht trin mnh, s
lng t bo vi khun > 107 CFU/ml trong cc khong thi gian kho st m bo
probiotic cung cp cho c th l .
4.3. Kho st qu trnh lm men ci.
Bng 4.4. Kt qu pH men vi khun L. acidophilus v L. bulgaricus qua thi gian
kho st
Thi gian
(gi)
0
2
4
6
8
10
pH men vi khun
L. acidophilus
6,33
6,23
6,09
5,98
5,67
5,14
pH men vi khun
L. bulgaricus
6,34
6,2
6,13
5,9
5,64
5,15
7
6
pH
5
4
pH men L.
acidophilus
pH men L.
bulgarucus
2
1
0
0
10
Thi gian( gi)
Hnh 4.7: pH men vi khun L. acidophilus v L. bulgaricus qua

thi gian kho st
38
Bng 4.5: M t cm quan men vi khun qua thi gian kho st

Thi gian
Men vi khun
Men vi khun
( gi)
L. acidophilus
L. bulgaricus
0
2
4
Cu trc lng
Cu trc lng
Mu trng c
Mu trng c
Mi u v mi bo
Mi u v mi bo
Cha ng, dng lng
Cha ng, dng lng
Cu trc st di y h, mi
Cha ng, c mi chua thong
u nh, v hi chua v bo
v mi chua nh
nhiu.
6
8
10
Cu trc sn phm c li, b
Cu trc sn phm st, khng
mt mn, c v chua kt hp vi
c, c mi sa chua mnh, v
v u.
hi chua kt hp vi v u.
Sn phm b tch mt lp nc
Cu trc sn phm c li, mi
trn b mt.
chua v c mi u nnh b h.
Sn phm b tch nc hon
Sn phm b tch nc hon
ton.
ton.
Bng nh gi cm quan, chng ti nhn thy i vi men L. acidophilus ti

thi im 6 gi t v mt gi tr cm quan cu trc v mi, nhng v v th khng
chua lm ( pH=5,98). Cn i vi men vi khun L. bulgaricus, ti thi im 6 gi
cm quan v cu trc khng t nhng t v mi, do vi khun ny c tc dng sinh
hng trong sn xut sa chua.
Kt lun:
Kt hp cm quan v pH sn phm, ng cong sinh trng ca vi khun trong
mi trng dch sa ( 4.2), chng ti chn men ci cho ng dng vo sn xut sa
chua l men vi khun ti 6 gi .
4.4. Kho st qu trnh ln men
Qu trnh ln men sa chua s b tc ng bi nhiu yu t. Trong ti ny
chng ti tin hnh kho st cc yu t sau: thi gian ln men, t l phi ch gia sa
39
u nnh v sa ti, t l ging cy ( t l gia L. acidophilus v L. bulgaricus), hm

lng ng saccharose b sung.
4.4.1. Kho st thi gian ln men
Bng 4.6: Kt qu pH ca dch sa theo thi gian ln men
Thi gian ( gi)
PH
6.45
6.3
5.92
5.36
5.02
4.73
4.63
7
6
5
4
pH
3
2
1
0
0
Thi gian( gi)
Hnh 4.8: Kt qu pH ca dch sa theo thi gian ln men
Bng 4.6: M t cm quan dch sa theo thi gian ln men

Thi gian( gi)
0
2
3
4
5
c im sn phm
Cha ng, dng lng
Cu trc bt u st di y h, mu xanh nht ca
to, mi chua nh, v u v ngt nhiu.
Cu trc sn phm c li, b mt lng, mu xanh
nht, mi chua mnh, v chua nh,v u v bo nhiu.
Sn phm b tch mt lp nc b mt, mu xanh
nht, mi chua mnh, v chua nh, v u v bo nhiu
Sn phm b tch nc trn b mt v di y h,
mi chua mnh kt hp vi mi i ca u nnh, v
40
chua v v u mnh, v bo gim.

6
Sn phm b tch nc hon ton, mi i chua ca u

nnh.
Bng nh gi cm quan chng ti nhn thy sn phm sau 3 v 4 gi ln men

c cm quan v mi v v ging nhau nhng khc nhau v cu trc sn phm. Kt hp
3 yu t: pH, cm quan v ng cong sinh trng ca vi khun trong mi trng dch
sa chng ti chn thi gian ln men sn phm l 3 gi.
Theo Gloria v cng s (2004), khi b sung thm sinh khi to S. platensis vo
sa s thc y qu trnh ln men lactic. Do hi gian ln men ca sa chua u
nnh to spirulina ( 3 gi) ngn hn so vi thi gian ln men ca cc sn phm sa
chua khc ( 4 - 6 gi).
Mt khc, pH ca sn phm sa chua u nnh to spirulina ( 5,36) l cao hn
nhiu so vi cc sn phm sa chua thng thng ( 4,0 - 4,6), chng ti chp nhn gi
tr pH ny v nu pH ca sn phm trong qu trnh kho st ny nu < 5 th sn phm
s khng t v mt cu trc. L do lm xy ra hin tng ny c th c gii thch
nh sau: pH ng in ca protein u nnh nm trong khong 4,2 - 4,6, kh nng ht
nc ca protein l thp nht ti pH ng in ca chng, nn pH ca sn phm cng
tin v gi tr pH ng in ny th sn phm cng b tch nc.
4.4.2. Kho st t l phi ch gia sa u nnh v sa ti tit trng khng
ng
Nhm lm gim bt mi u nnh, chng ti tin hnh kho st t l phi ch
gia sa u nnh v sa ti tit trng vi cc t l nh sau
D: t l 4 phn sa u nnh v 6 phn sa ti
E: t l 6 phn sa u nnh v 4 phn sa ti
F: t l 8 phn sa u nnh v 2 phn sa ti
T kt qu nh gi cm quan ph lc 2.1, v kt qu phn tch thng k ph
lc 3.1 chng ti rt ra c Bng 4.7
41
Bng 4.7: im trung bnh kt qu nh gi cm quan sn phm khi thay i t

l phi ch gia u nnh v sa ti
T l phi ch
Kt qu nh gi cm quan
71
63
46
-0,31a
-0,03a
0,39b
Tng im theo th t so hng

im trung bnh cm quan ( 0,32)
Ghi ch: Trong cng mt hng, cc s mang cng ch ci th khc bit khng
c ngha tin cy 95% theo trc nghim LSD ( P>0,05).
Qua Bng 4.7 chng ti c nhn xt v kt lun nh sau
T l phi ch gia sa u nnh v sa ti khc nhau nh hng c ngha
n kt qu nh gi cm quan ca sn phm tin cy 95% ( P<0,05). V nghim
thc F( t l 8 phn sa u nnh v 2 phn sa ti) c im trung bnh cm quan
cao nht v khc bit c ngha tin cy 95% theo trc nghim LSD ( P<0,05) so
vi 2 nghim thc cn li nn chng ti chn nghim thc F ( t l 8 phn sa u
nnh v 2 phn sa ti) lm thng s c nh cho cc th nghim tip theo.
4.4.3. Kho st hm lng ng saccharose b sung
Hm lng ng b sung nh hng n qu trnh ln men v v ca sn
phm, trong nghin cu ny chng ti nghin cu nh hng ca hm lng ng b
sung n cm quan ca sn phm. Chng ti tin hnh b sung nh sau
G: hm lng ng b sung l 6% (w/v)
H: hm lng ng b sung l 8% (w/v)
I: hm lng ng b sung l 10% (w/v)
Bng 4.8: im trung bnh kt qu nh gi cm quan sn phm khi thay i
hm lng ng b sung
Hm lng ng b sung
Kt qu nh gi cm quan
42
72
42
66
-0,34a
0,51b
-0,17a
Hm lng ng b sung vo sa khc nhau nh hng c ngha n kt
qu nh gi cm quan ca sn phm tin cy 95% (P<0,05). Nghim thc H (
hm lng ng b sung vo l 8%) c im trung bnh cm quan cao nht v khc
bit c ngha tin cy 95% theo trc nghim LSD ( P<0,05) so vi 2 nghim thc
cn li nn chng ti chn nghim thc F ( b sung 8% ng) lm thng s c nh
cho cc th nghim tip theo.
4.4.4. Kho st t l ging cy
Chng ti tin hnh thay i t l ging cy ( t l gia L. acidophilus v L.
bulgaricus) nh sau
J: 4% ging L. acidophilus v 6% ging L. bulgaricus trong 10% ging cy vo
sa ( v/v)
K: 5% ging L. acidophilus v 5% ging L. bulgaricus trong 10% ging cy
vo sa ( v/v)
L: 6% ging L. acidophilus v 4% ging L. bulgaricus trong 10% ging cy
vo sa ( v/v)
Bng 4.9: im trung bnh kt qu nh gi cm quan sn phm khi thay i t
l ging cy
T l ging cy
Kt qu nh gi cm quan
60
63
57
-0,56a
-0,85a
0,85a
43
T l ging cy vo sa ln men nh hng khng c ngha n kt qu

nh gi cm quan sn phm tin cy 95% ( P<0,05). Tuy nhin nghim thc L
(6% ging L. acidophilus v 4% ging L. bulgaricus) c im trung bnh cm quan
cao nht nn chng ti chn nghim thc L lm thng s c nh cho cc th nghim
tip theo.
4.5. Kho st hm lng to b sung
Nhm kho st nh hng ca hm lng to b sung n gi tr cm quan ca
sn phm, chng ti tin hnh b sung to vo sa vi cc nghim thc nh sau
A: hm lng to b sung 6% ( v/v)
B: hm lng to b sung 8% ( v/v)
C: hm lng to b sung 10% ( v/v)
Bng 4.10: im trung bnh kt qu nh gi cm quan sn phm khi thay i
hm lng to b sung
Hm lng to b sung
Kt qu nh gi cm quan
44
60
76
0,45a
0b
-0,45c
Ghi ch: Trong cng mt hng, cc s mang cc ch ci khc nhau th khc

bit c ngha tin cy 95% theo trc nghim LSD ( P>0,05).
Hm lng to b sung vo sa ln men nh hng c ngha n kt qu
nh gi cm quan sn phm tin cy 95% ( P< 0,05). V nghim thc A(hm
lng to b sung 6%) c im trung bnh cm quan cao nht v khc bit c ngha
tin cy 95% theo trc nghim LSD ( P<0,05) so vi 2 nghim thc cn li nn
chng ti chn nghim thc A lm thng s c nh cho cc th nghim tip theo.
44
4.6. Kho st thi gian bo qun sn phm

Chng ti tin hnh bo qun sn phm 4OC trong thi gian 16 ngy. Trong
thi gian bo qun chng ti theo di cc ch tiu pH, acid v s lng vi khun
lactic trong sn phm v th c kt qu nh sau
Bng 4.11: Kt qu kho pH, acid v s lng vi sinh vt theo thi gian bo
qun
Thi gian ( ngy)
Log s lng vi
sinh vt (CFU/ml)
pH
acid ( %)
8,18
5,2
0,28
8,53
5,13
0,3
9,08
0,31
9,64
4,9
0,35
9,83
4,88
0,4
10
9,85
4,84
0,42
12
9,91
4,8
0,5
14
9,8
4,75
0,53
16
9,7
4,7
0,59
45
11
5,3
10
5,2
5,1
7
6
4,9
Log s lng
(CFU/ml)
4,8
pH
4,7
4,6
4,5
1
0
4,4
0
10
12
14
16
Thi gian( ngy)
Hnh 4.12: Kt qu PH v s lng vi khun lactic theo thi

gian bo qun sn phm
acid(%)
0,7
0,6
0,5
0,4
acid(%)
0,3
0,2
0,1
0
0
10
15
20
Thi gian ( ngy)
Hnh 4.13: Kt qu acid ca sn phm theo thi gian bo

qun
Qua Bng 4.11, Hnh 4.12 v Hnh 4.13 chng ti c nhn xt nh sau
S lng vi khun lactic trong sn phm l trn 108 CFU/ml trong thi gian
chng ti kho st, m bo s lng vi khun probiotic cung cp cho c th ( > 107
CFU/ml).
46
pH ca sn phm c s gim mnh trong 8 ngy u bo qun, ng thi l s

tng ln ca acid, c th gii thch iu ny nh sau: bo qun sn phm 4OC c
tc dng hn ch tc sinh trng v pht trin ca vi khun ch khng kim hm
hon ton s pht trin ca chng, mt khc vi khun cng c s thch nghi vi iu
kin mi trng mi nn s lng t bo ca chng tng ln, lng acid lactic sinh ra
trong qu trnh trao i cht ca vi chng lm gim pH sn phm v lm tng acid.
Tuy nhin, do pH ng in ca protein u nnh khong 4,2 - 4,6, quan st
cm quan sn phm trong qu trnh bo qun chng ti nhn thy sau 10 ngy bo
qun sn phm c hin tng tch nc b mt. Do chng ti chn thi gian bo
qun ti u cho sn phm l 10 ngy.
Nhm xc nh mc chp nhn ca ngi tiu dng i vi sn phm sa
chua u nnh to spirulina, chng ti tin hnh cho nh gi cm quan theo phng
php cho im th hiu gia 2 mu nh sau
Mu 206: sa chua u nnh to spirulina
Mu 207: sa chua Vinamilk
Kt qu nh gi th hiu 30 ngi th c th hin Bng 4.12
Bng 4.12: im nh gi th hiu trung bnh ca ngi th cho cc c tnh ca
sn phm
Ch tiu
Mu
Mu 206
Mu 207
a thch v a thch v a thch v a thch v

mu
mi
v
cu trc
7,5
7,8
6,8
7,47
6,73
8,13
47
7,03
8,17
a thch
chung
7,03
7,97
9
8
im trung bnh
7
6
5
Mu 206
Mu 207
3
2
1
0
Mu sc
Mi
Cu trc
Chung
Ch tiu cm quan
Hnh 4.14: im nh gi th hiu ca ngi th cho hai sn

phm
i vi sn phm nghin cu, ch tiu v mu c ngi th a thch nht v
thp nht l ch tiu v mi v v v.
im trung bnh i vi cc ch tiu ca sn phm nghin cu dao ng trong
khong t 6 - 7. Nh vy, mc a thch ca ngi th i vi mu nghin cu
trong khong hi thch n thch. Ch tiu v mi v v l thp nht, thp hn nhiu so
vi sn phm sa chua trn th trng, iu ny c th l do sn phm vn cn mi
u v v cha chua lm, vi khun L. bulgaricus to c cc hp cht hng cha
kh c mi u hon ton, c tnh ca protein u nnh s b tch nc nu sn
phm chua hn, do cn nghin cu loi cht n nh thch hp lm cho sn phm
nghin cu c ci thin v v hn.
Nhn chung, c hai sn phm u t im trn 6 c ngha l ngi th cng
u chp nhn c sn phm ang nghin cu v sn phm ang c yu thch trn th
trng.
48
4.7. Kt qu phn tch cc ch tiu vi sinh v ha hc

4.7.1. Kt qu phn tch ch tiu vi sinh
Bng 4.13: Kt qu kim tra ch tiu vi sinh sn phm
Ch tiu
Tng s vi khun hiu kh
Tng s nm men, nm mc
n v
Mc cho php
Kt qu kim tra
Coliforms
E.coli
4.7.2. Kt qu phn tch ch tiu ha hc
Bng 4.14: Kt qu kim tra cc ch tiu ha hc
Ch tiu
n v
Kt qu kim tra
m tng s
ng tng s
Cht bo
pH
acid
49
Chng 5
KT LUN V NGH
5.1. Kt lun
Qua qu trnh tin hnh th nghim, chng ti rt ra nhng kt lun sau
3 Trong mi trng MRS broth, sinh khi vi khun Lactobacillus acidophilus
t cc i sau 15 gi nui cy, sinh khi vi khun Lactobacillus bulgaricus t cc
i sau 18 gi nui cy.
3 Trong mi trng sa, sinh khi vi khun Lactobacillus acidophilus v
Lactobacillus bulgaricus t cc i sau 15 gi nui cy.
3 Thi gian thu men ci vi khun l 6 gi .
3 Cc thng s ca qu trnh ln men
# T l phi ch gia sa u nnh v sa ti UHT l 8:2
# Thi gian ln men 3 gi, 43OC
# Cy 10% men ci vi t l ging cy gia Lactobacillus acidophilus v
Lactobacillus bulgaricus l 6:4
# Hm lng ng saccharose b sung l 8%
# Hm lng to b sung l 6%, gelatin 5%
3 Thi gian bo qun sn phm ti u l 10 ngy 4OC.
5.2. ngh
Do hn ch v thi gian cng nh trang thit b nn chng ti khng th kho
st ht cc th nghim c lin quan cho sn phm c cht lng tt hn nn chng
ti c mt s ngh sau
3 Theo di nht sn phm theo thi gian ln men
3 Nghin cu cht n nh b sung thch hp sn phm t ci thin hn v
v v cu trc.
3 Kho st thm mt s hng liu b sung v sa chua ci thin mi u
nnh.
50
TI LIU THAM KHO

TING VIT
1) L Hong , Cy u nnh, NXB Khoa hc k thut, 1997.
2) ng nh Kim, ng Hong Phc Hin, Cng ngh sinh hc vi to, NXB
Nng nghip, 1999.
3) L Vn Lng, Spirulina - Nui trng, S dng trong y dc v dinh dng,
NXB Y hc, 1999.
4) Trng Thanh Long, Gao trnh Bo qun v ch bin trng sa, tr201
5) Nguyn c Lng, Cng ngh vi sinh tp 2 Vi sinh vt hc cng nghip,
NXB i hc quc gia TP.HCM.
6) Nguyn c Lng, Cng ngh vi sinh - Tp3: Thc phm ln men truyn
thng, NXB i hc Quc gia TPHCM, 2006, trang 15-33.
7) Phm Vn Thiu, Cy u tng- k thut trng v ch bin sn phm, NXB
Nng nghip, H Ni.
8) Chu Th Thm, Phan th Li, Nguyn Vn T, Ga tr dinh dng v phng
php ch bin sa, NXB Lao ng, trang 5.
9) L Ngc T, Ha hc thc phm, NXB Khoa hc K thut, H Ni, 2001,
trang 42- 131.
10) H Duyn T v Lu Dun, nh gi thc phm bng phng php kim
nghim cm quan, NXB Khoa hc K thut, H Ni, 1998.
TING NC NGOI
11) C. r. Rekha, G. Vijayalakshmi, Biomolecules and Nutritional Quality of
Soymilk Fermented with Probiotic Yeast and Bacteria, Central Food
Technological Research Institute, 2008.
12) D. O. Otieno, J. F. Ashton, v Nagendra P. Shah, Stability of -glucosidase
Activity Produced by Bifidobacterium and Lactobacillus spp. in Fermented
Soymilk During Processing and Storage, Journal of Food Science, Vol. 70,
2005.
13) Dillon, J.C., Phuc, A.P., Dubacq, J.P., 1995. Nutritional value of the alga
Spirulina, World Rev.Nutr.Diet, Vol 77, 32-46.
51
14) Elina Tuomola, Ross Crittenden, Martin Playne, Erika Isolauri, and Seppo
Salminen, Quality assurance criteria for probiotic bacteria, The American
Journal of Clinical Nutrition, 2001.
15) Food and Agriculture Organization of the United Nations (FAO), Technology of
production edible flours and protein from soybeans Chapter 1: The soybean,
1999)
16) Gloria, Z.C., Josa, L.P., Mara, C.Z., Manica, M.S.C, 2004. Effect of S.platensis
biomass on the growth of lactic acid bacteria in milk, World Journal of
Microbiology and Biotech nology, Vol 16, 563-565.
17) Henrickson, R. 1989. Earth Food Spirulina. Laguna Beach, California: Ronore
Enterprises Inc, 2342.
18) Kalidas Shetty, Gopinadhan Paliyath, Anthony Pometto, Robert E. Levin, Food
Biotechnology, second Edition, Published by CRC Press, 2006.
19) KeShun Liu, Soybeans: Chemistry, Technology & Utilization, An Aspen
Publication, 1999.
20) Paul, P.B., Kirk, E.A., 1999. Commercial developments in microalgal
biotechnology, Microalgal biotechnology, Vol 35, 215-226.
21) Peter Golbitz and Joe Jordan, Soy Application in Food, Chapter 1- Soyfoods:
Market and Products, Published by CRC Press Taylor & Francis Group, 2006.
22) Tadao Saito, Selection of useful probiotic lactic acid bacteria from the
Lactobacillus acidophilus group and their applications to functional foods,
Animal Science Journal , 2004.
TI LIU INTERNET
23) www.soya.be
24) www.soyinfocenter.com/HSS/fermented_soymilk.php
25) www.mdidea.com/products/herbextract/soy/data.html
52
PH LC
PH LC 1: PHNG PHP PHN TCH CC CH TIU
1.1. nh lng vi khun bng phng php m khun lc
a) Nguyn tc
Cy mt th tch xc nh mu huyn ph vi sinh vt ln mi trng thch trong
hp petri. Mi trng c chn c thnh phn c cht c trng v thch hp cho loi
vi sinh vt cn nh lng pht trin. Sau mt khong thi gian nui cy, trn b mt
thch s xut hin cc khun lc v ta c th quan st c bng mt thng. Da vo
s khun lc m c, th tch mu cy v h s pha long, ta c th suy ra s
khun lc c trong 1ml mu kho st ban u.
u im ca phng php ny l ta ch nh lng nhng t bo sng c trong
mu kho st ban u. Tuy nhin, phng php ny cng c nhc im l thi gian
di, tn km chi ph ha cht.
b) Cch tin hnh
xc nh mt t bo trn mt n v th tch trong mi trng MRS
broth, hay sa chua u nnh ln men, chng ti tin hnh phng php cy gt theo
cc bc nh sau:
Bc 1: Chun b mu v mi trng thch.
Mi trng MRS agar gieo cy, nc pha long c pha ch theo cng
thc v hp tit trng 1210C trong 15 pht.
Tin hnh pha long mu vi cc pha long khc nhau
9 Dung dch mu 10-1: Ht 1ml dung dch mu lng cho vo 9ml nc
pepton. Lc u 2-3 pht, thu c dung dch mu 10-1.
9 Dung dch mu 10-2, 10-3, 10-4 : Ht chnh xc 1 ml dung dch mu th
10-1 cho sang ng nghim cha sn 9 ml nc pepton. Lc u trong 2-3 pht, thu
c dung dch 10-2. Tip tc lm tng t nh vy, ta thu c cc dung dch mu
tng ng 10-3, 10-4 ..
53
Bc 2: Nui cy mu
Ghi k hiu mu v nng dung dch mu th ln a thch MRS.

Chn 2 hoc 3 nng pha long lin tip m d kin cha s khun lc nm
trong khong 25 250 khun lc trn mi a. Mi bc pha long dng micropipet v
trng ht chnh xc 0,1ml nh ln b mt a thch MRS c hong kh. Dng que
trang v trng dn u mu trn b mt thch. Tin hnh cy mu mi pha
long vi hai a thch ging nhau (lp li hai ln).
Tip theo, ch cho b mt thch kh, tin hnh bao hp petri, lt p hp v t
chng vo t m. Nui cy 370C trong thi gian t 2 ngy n 3 ngy.
c) Tnh kt qu
Kt thc thi gian , m s khun lc pht trin trn mi trng thch mi
a.
Mt t bo c tnh theo cng thc sau:
Trong :
C (CFU/ml): s t bo vi khun trong 1 ml mu.

N (CFU): tng s khun lc m c trn cc a.
ni: s a cy nng pha long th i.
Vi (ml): th tch mu cy trong mi a.
fi: pha long th i.
1.2. Phng php xc nh ng cong sinh trng

54
a) Nguyn tc
Da vo s pht trin ca vi khun cc thi im khc nhau
b) Cch tin hnh
Xy dng ng cong sinh trng ca Lactobacillus acidophilus,
Lactobacillus bulgaricus trong mi trng MRS.
9 Vi khun c nui trong mi trng MRS broth vi t l ging cy l
10% 370C.
9 Khong 3 gi ly mu cy trang trn mi trng MRS agar.
9 Dng phng php m khun lc xc nh mt t bo (CFU/ml) c
trong mu.
9 V th ng cong sinh trng gia log cfu/ml trc tung v thi
gian trc honh.
Xy dng ng cong sinh trng ca Lactobacillus acidophilus,
Lactobacillus bulgaricus trong mi trng sa u nnh v sa ti.
9 Vi khun Lactobacillus acidophilus c nui trong mi trng sa u
nnh vi t l ging cy c chn v nhit c kho st.
9 Khong 5 gi ly ra cy trang trn mi trng MRS agar.
9 Dng phng php m khun lc xc nh mt t bo (CFU/ml)
trong trong mu ging ban u.
9 V th ng cong sinh trng gia log(CFU/mL) trc tung v thi
gian trc honh.
1.3. Phiu nh gi cm quan
1.3.1.Php th so hng:
PHIU NH GI CM QUAN
Php th so hng
Tn sn phm: Sa chua u nnh to spirulina
H v tn:....................................
Ngy th: ..........................................
Bn nhn c 3 mu sa u nnh k hiu 123, 132, 321. Bn hy th nm cc mu

v sp xp mi v theo mc a thch. Mu yu thch nht s xp v tr th nht v
ght nht s xp v tr cui cng.
Mu:
123
132
321
55
Th t yu thch:
.....
......
......
Bnh lun:
...........................................................................................................................................
..............................................................................................................
Cm n s ng gp kin ca cc bn!
1.3.2 .Php th cho im th hiu
PHIU NH GI CM QUAN
Php th cho im th hiu
Tn sn phm: Sa chua
H v tn:....................................
Ngy th: ..........................................
Bn nhn c 2 mu sa chua u nnh to spirulina k hiu 206, 207. Bn hy th

nm v cho im theo thang im sau:
1: Rt ght
2: Kh ght
3: Ght
4: Hi ght
6: Hi thch
7: Thch
8: Kh thch
9: Rt thch
5: Khng ght, khng thch

Mu sc
Mu
206
207
206
207
206
207
206
207
im
Mi
Mu
im
V
Mu
im
Trng thi
Mu
im
56
Mc a thch chung
Mu
206
207
im
Bnh lun:
...........................................................................................................................................
..............................................................................................................
Cm n s ng gp kin ca cc bn!
PH LC 2: KT QU NH GI CM QUAN
2.1. Kt qu nh gi cm quan th nghim t l phi ch gia sa u nnh v
sa ti
Hng
im
Cm quan
Mu
Mu
Mu
vin
Mu 4:6 Mu 6:4 Mu 8:2
4:6
6:4
8:2
1
3
1
2
-0,85
0,85
0
2
3
2
1
-0,85
0
0,85
3
2
3
1
0
-0,85
0,85
4
3
2
1
-0,85
0
0,85
5
2
3
1
0
-0,85
0,85
6
3
1
2
-0,85
0,85
0
7
3
2
1
-0,85
0
0,85
8
1
3
2
0,85
-0,85
0
9
3
2
1
-0,85
0
0,85
10
2
3
1
0
-0,85
0,85
11
3
1
2
-0,85
0,85
0
12
1
3
2
0,85
-0,85
0
13
3
1
2
-0,85
0,85
0
14
3
2
1
-0,85
0
0,85
15
3
2
1
-0,85
0
0,85
16
2
3
1
0
-0,85
0,85
17
3
1
2
-0,85
0,85
0
18
3
2
1
-0,85
0
0,85
19
1
2
3
0,85
0
-0,85
20
3
2
1
-0,85
0
0,85
21
1
3
2
0,85
-0,85
0
22
3
1
2
-0,85
0,85
0
23
1
3
2
0,85
-0,85
0
57
24
25
26
27
28
29
30
Tng im
1
3
1
3
2
3
3
71
3
2
2
2
3
1
2
63
2
1
3
1
1
2
1
46
0,85
-0,85
0,85
-0,85
0
-0,85
-0,85
-0,85
0
0
0
-0,85
0,85
0
0
0,85
-0,85
0,85
0,85
0
0,85
2.2. Kt qu nh gi cm quan th nghim kho st hm lng ng b sung

Cm quan
vin
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
Mu 6%
3
2
3
3
3
1
2
2
3
3
3
1
1
3
3
2
2
1
3
3
2
1
3
3
3
3
3
Hng
Mu 8%
1
1
2
1
1
2
3
1
1
1
2
2
2
1
1
1
1
2
2
1
1
2
1
1
1
2
2
Mu 10%
2
3
1
2
2
3
1
3
2
2
1
3
3
2
2
3
3
3
1
2
3
3
2
2
2
1
1
58
Mu 6%
-0,85
0
-0,85
-0,85
-0,85
0,85
0
0
-0,85
-0,85
-0,85
0,85
0,85
-0,85
-0,85
0
0
0,85
-0,85
-0,85
0
0,85
-0,85
-0,85
-0,85
-0,85
-0,85
im
Mu 8%
0,85
0,85
0
0,85
0,85
0
-0,85
0,85
0,85
0,85
0
0
0
0,85
0,85
0,85
0,85
0
0
0,85
0,85
0
0,85
0,85
0,85
0
0
Mu 10%
0
-0,85
0,85
0
0
-0,85
0,85
-0,85
0
0
0,85
-0,85
-0,85
0
0
-0,85
-0,85
-0,85
0,85
0
-0,85
-0,85
0
0
0
0,85
0,85
28
29
30
Tng im
2
2
3
72
1
1
1
42
3
3
2
66
0
0
-0,85
0,85
0,85
0,85
-0,85
-0,85
0
2.3. Kt qu nh gi cm quan th nghim kho st t l ging cy

Cm quan
vin
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
Mu
4a:6b
3
2
1
1
3
2
1
2
3
1
3
2
1
3
2
3
1
3
2
1
3
1
2
2
1
2
3
2
3
1
Hng
Mu
5a:5b
2
3
2
3
2
3
2
3
2
2
2
3
2
2
3
1
2
2
3
2
1
2
3
1
2
1
1
1
2
3
Mu
6a:4b
1
1
3
2
1
1
3
1
1
3
1
1
3
1
1
2
3
1
1
3
2
3
1
3
3
3
2
3
1
2
59
Mu
4a:6b
-0,85
0
0,85
0,85
-0,85
0
0,85
0
-0,85
0,85
-0,85
0
0,85
-0,85
0
-0,85
0,85
-0,85
0
0,85
-0,85
0,85
0
0
0,85
0
-0,85
0
-0,85
0,85
im
Mu
5a:5b
0
-0,85
0
-0,85
0
-0,85
0
-0,85
0
0
0
-0,85
0
0
-0,85
0,85
0
0
-0,85
0
0,85
0
-0,85
0,85
0
0,85
0,85
0,85
0
-0,85
Mu
6a:4b
0,85
0,85
-0,85
0
0,85
0,85
-0,85
0,85
0,85
-0,85
0,85
0,85
-0,85
0,85
0,85
0
-0,85
0,85
0,85
-0,85
0
-0,85
0,85
-0,85
-0,85
-0,85
0
-0,85
0,85
0
Tng im
60
63
57
2.4. Kt qu nh gi cm quan th nghim kho st hm lng to b sung

Cm quan
vin
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
Tng im
Mu
6%to
1
1
2
2
1
3
1
1
2
2
1
1
1
2
1
1
1
3
1
2
1
1
1
2
2
1
1
1
1
3
44
Hng
Mu
8%to
2
3
1
1
2
2
2
3
1
1
2
3
3
1
2
3
2
1
2
1
3
2
2
1
3
2
2
3
2
2
60
Mu
10%to
3
2
3
3
3
1
3
2
3
3
3
2
2
3
3
2
3
2
3
3
2
3
3
3
1
3
3
2
3
1
76
Mu
6%to
0,85
0,85
0
0
0,85
-0,85
0,85
0,85
0
0
0,85
0,85
0,85
0
0,85
0,85
0,85
-0,85
0,85
0
0,85
0,85
0,85
0
0
0,85
0,85
0,85
0,85
-0,85
im
Mu
8%to
0
-0,85
0,85
0,85
0
0
0
-0,85
0,85
0,85
0
-0,85
-0,85
0,85
0
-0,85
0
0,85
0
0,85
-0,85
0
0
0,85
-0,85
0
0
-0,85
0
0
2.5. Kt qu nh gi cm quan th hiu sn phm sa chua u nnh to

spirulina v sa chua Vinamilk
60
Mu
10%to
-0,85
0
-0,85
-0,85
-0,85
0,85
-0,85
0
-0,85
-0,85
-0,85
0
0
-0,85
-0,85
0
-0,85
0
-0,85
-0,85
0
-0,85
-0,85
-0,85
0,85
-0,85
-0,85
0
-0,85
0,85
Cm
quan vin
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
im
trung
bnh
Sa chua u nnh to spirulina (206)

Mu
Cu
Chung
Mi V
sc
trc
6
5
6
7
6
8
5
7
6
7
8
7
7
8
7
7
6
8
9
7
8
7
8
7
7
9
7
5
7
7
8
8
7
8
8
7
6
7
7
7
9
7
5
7
7
8
7
5
8
8
7
8
6
5
6
8
7
6
6
7
8
7
8
8
7
7
8
6
6
6
8
7
7
8
7
8
7
7
6
7
8
7
7
8
8
7
7
7
8
7
7
5
6
5
6
8
6
7
6
7
7
7
8
7
7
7
6
6
7
7
8
7
7
8
8
8
8
8
8
8
5
7
5
7
6
7
8
8
7
8
8
7
8
6
7
7
8
6
7
7
6
5
6
7
6
8
7
8
7
8
7,5
6,8
6,73
7,03
7,03
Sa chua Vinamilk (207)

Mu
Cu
Mi V
Chung
sc
trc
7
7
8
8
8
7
6
9
9
8
8
7
9
9
8
7
8
8
8
8
9
8
9
9
9
9
8
9
9
9
8
7
8
8
7
7
8
8
9
8
9
8
9
8
8
8
8
9
9
8
7
7
7
7
7
8
7
9
7
8
9
8
9
8
9
7
8
7
8
8
8
8
8
8
8
9
7
6
9
8
7
8
8
8
8
9
7
7
9
8
9
8
9
7
8
8
8
9
7
8
7
8
8
9
8
8
8
9
9
9
7
8
8
9
8
8
5
5
8
5
6
7
8
7
7
8
7
9
7
8
8
8
8
8
8
7
8
8
8
8
7
7
8
8
8
8
7
8
8
9
7,8
7,47 8,13 8,17
7,97
PH LC 3: KT QU PHN TCH THNG K

3.1. Kt qu phn tch thng k nh gi cm quan th nghim t l phi ch gia
sa u nnh v sa ti
61
3.1.1.Bng ANOVA
One-Way Analysis of Variance
------------------------------------------------------------------------------Data: CQTYLE.Diem
Level codes: CQTYLE.Mau
Labels:
Means plot: LSD
Confidence level: 95
Range test: LSD
Analysis of variance
------------------------------------------------------------------------------Source of variation
Sum of Squares
d.f.
Mean square
F-ratio Sig.
level
------------------------------------------------------------------------------Between groups
7.626389
2
3.8131944
9.295
.0002
Within groups
35.691500
87
.4102471
------------------------------------------------------------------------------Total (corrected)
43.317889
89
0 missing value(s) have been excluded.
3.1.2. Bng trung bnh nghim thc

Table of means for CQTYLE.Diem by CQTYLE.Mau
------------------------------------------------------------------------------Stnd. Error Stnd. Error
95 % LSD
Level
Count
Average
(internal)
(pooled s)
intervals for
mean
------------------------------------------------------------------------------D
30
-.3116667
.1319544
.1169397
-.4760567
.1472766
E
30
-.0283333
.1187019
.1169397
-.1927234
.1360567
F
30
.3966667
.0975838
.1169397
.2322766
.5610567
------------------------------------------------------------------------------Total
90
.0188889
.0675152
.0675152
-.0760218
.1137995
3.1.3. Bng so snh s khc bit gia cc nghim thc

Multiple range analysis for CQTYLE.Diem by CQTYLE.Mau
------------------------------------------------------------------------------Method: 95 Percent LSD
Level
Count
Average
Homogeneous Groups
------------------------------------------------------------------------------D
30
-.3116667
X
62
E
30
-.0283333
X
F
30
.3966667
X
------------------------------------------------------------------------------contrast
difference
limits
D - E
-0.28333
0.32878
D - F
-0.70833
0.32878 *
E - F
-0.42500
0.32878 *
------------------------------------------------------------------------------* denotes a statistically significant difference.
3.2. Kt qu phn tch thng k nh gi cm quan th nghim kho st hm

lng ng b sung
3.2.1.Bng ANOVA
------------------------------------------------------------------------------Data: CQDUONGS.Diem
Level codes: CQDUONGS.Mau
Labels:
Means plot: LSD
Range test: LSD
Sum of Squares
d.f.
Mean square
F-ratio Sig.
level
------------------------------------------------------------------------------Between groups
12.138000
2
6.0690000
16.917
.0000
Within groups
31.212000
87
.3587586
------------------------------------------------------------------------------Total (corrected)
43.350000
89

Table of means for CQDUONGS.Diem by CQDUONGS.Mau
95 % LSD
Level
Count
Average
(internal)
(pooled s)
intervals for
mean
------------------------------------------------------------------------------G
30
-.3400000
.1195153
.1093555
-.4937283
.1862717
63
H
30
.5100000
.0874084
.1093555
.3562717
.6637283
I
30
-.1700000
.1181174
.1093555
-.3237283
.0162717
------------------------------------------------------------------------------Total
90
.0000000
.0631364
.0631364
-.0887551
.0887551

Multiple range analysis for CQDUONGS.Diem by CQDUONGS.Mau
Level
Count
Average
Homogeneous Groups
------------------------------------------------------------------------------G
30
-.3400000
X
I
30
-.1700000
X
H
30
.5100000
X
------------------------------------------------------------------------------contrast
difference
limits
G - H
-0.85000
0.30746 *
G - I
-0.17000
0.30746
H - I
0.68000
0.30746 *
------------------------------------------------------------------------------* denotes a statistically significant difference
3.3. Kt qu nh gi cm quan th nghim kho st t l ging cy

3.3.1.Bng ANOVA
------------------------------------------------------------------------------Data: CQGIONG.Diem
Level codes: CQGIONG.Mau
Labels:
Means plot: LSD
Range test: LSD
Sum of Squares
d.f.
Mean square
F-ratio Sig.
level
------------------------------------------------------------------------------Between groups
.497722
2
.2488611
.506
.6049
Within groups
42.820167
87
.4921858
------------------------------------------------------------------------------Total (corrected)
43.317889
89
64

Table of means for CQGIONG.Diem by CQGIONG.Mau
95 % LSD
Level
Count
Average
(internal)
(pooled s)
intervals for
mean
------------------------------------------------------------------------------J
30
-.0566667
.1284464
.1280867
-.2367267
.1233934
K
30
-.0850000
.1104887
.1280867
-.2650600
.0950600
L
30
.0850000
.1432214
.1280867
-.0950600
.2650600
------------------------------------------------------------------------------Total
90
-.0188889
.0739509
.0739509
-.1228466
.0850688

Multiple range analysis for CQGIONG.Diem by CQGIONG.Mau
Level
Count
Average
Homogeneous Groups
------------------------------------------------------------------------------K
30
-.0850000
X
J
30
-.0566667
X
L
30
.0850000
X
------------------------------------------------------------------------------contrast
difference
limits
J - K
0.02833
0.36012
J - L
-0.14167
0.36012
K - L
-0.17000
0.36012
3.4. Kt qu phn tch thng k nh gi cm quan th nghim kho st hm

lng to b sung
3.4.1. Bng ANOVA
------------------------------------------------------------------------------Data: CQTAO.Diem
Level codes: CQTAO.Mau
Labels:
65
Means plot: LSD
Range test: LSD
Sum of Squares
d.f.
Mean square
F-ratio Sig.
level
------------------------------------------------------------------------------Between groups
12.330667
2
6.1653333
17.292
.0000
Within groups
31.019333
87
.3565441
------------------------------------------------------------------------------Total (corrected)
43.350000
89

Table of means for CQTAO.Diem by CQTAO.Mau
95 % LSD
Level
Count
Average
(internal)
(pooled s)
intervals for
mean
------------------------------------------------------------------------------A
30
.4533333
.1057522
.1090174
.3000802
.6065865
B
30
.0000000
.1152708
.1090174
-.1532531
.1532531
C
30
-.4533333
.1057522
.1090174
-.6065865
.3000802
------------------------------------------------------------------------------Total
90
.0000000
.0629412
.0629412
-.0884807
.0884807

Multiple range analysis for CQTAO.Diem by CQTAO.Mau
Level
Count
Average
Homogeneous Groups
------------------------------------------------------------------------------C
30
-.4533333
X
B
30
.0000000
X
A
30
.4533333
X
------------------------------------------------------------------------------contrast
difference
limits
A - B
0.45333
0.30651 *
A - C
0.90667
0.30651 *
B - C
0.45333
0.30651 *
66

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NGHIN CU QUY TRNH SN XUT SA CHUA U

Kha lun c trnh p ng yu cu cp bng k s ngnh

Gio vin hng dn:

2.3.2.1. Lactobacillus acidophilus ............................................................................20

2.6.3. Phng php chun acid.............................................................................32

DANH SCH CC BNG

DANH SCH CC HNH

im vt tri c bit v gi tr dinh dng v c cha nhiu hp cht c gi tr trong

2.1.3.2.Thnh phn ha hc ca ht u nnh

Loi acid amin

Da vo chc nng sinh hc thc vt, protein trong ht u nnh c chia

Hnh 1. 1: nh hng ca pH n kh nng ha tan ca protein u nnh

chng li s hp th cc cht m qua mng rut, do khng nn n u nnh sng.

3Tempeh: l dng bnh c to thnh bi s nu u v trn vi vi khun, lo

Hnh 2.2: S vng i ca Spirulina [Ciferri, 1985]

S lng (% cht kh)

0,1% cht kh) cao hn 20 ln so vi trong carrot. Thnh phn cc vitamin ca

Spirulina giu st v calcium, h tr tt cho mu, cho xng v rng. Lng

Nhu cu hng ngy

% so vi nhu cu hng ngy

Spirulina cha 18 trong s 20 loi amino acid c bit (Fox, 1986). Mt s

Spirulina c mu xanh lam-lc l do Spirulina cha nhiu sc t vi hm lng

Bng 2.8: Cc cht mu trong Spirulina [ 5

Hm lng trong 10g

2.2.3.4.Tnh hnh sn xut, tiu th v ng dng ca to spirulina

2.3. Tng quan v vi sinh vt

2.3.2.1. Lactobacillus acidophilus [ 22]

2.4. Tng quan v sa v cht n nh

3Mi trng MRS agar (

3Mi trng MRS broth

3My xay sinh t

3Bnh tam gic c chia

3Dung dch NaOH 0,1N

thnh t sy) ca cng

3Ni autoclave SA 252F

2.4.Quy trnh ch bin sa chua u nnh to Spirulina

2.4.2.Gii thch quy trnh

Lm thun v gi ging c tin hnh nh sau: Vi khun trong ng ging mua

Cho ging vi khun lm

Xy dng ng cong sinh

Xy dng ng cong sinh

Kho st hm lng to b sung

Kho st thi gian bo qun sn

2.5.1.Xy dng ng cong tng trng ca Lactobacillus acidophilus,

c/Ch tiu theo di

c/Ch tiu theo di

d/ Phng php thc hin

2.5.4. Kho st qu trnh ln men.

2.5.4.4.Kho st t l ging cy.

2.6. Phng Php Phn Tch Cc Ch Tiu

C (CFU/g) l s t bo vi khun trong 1g mu

Hnh 4.1: L. acidophilus

Hnh 4.2: L. bulgaricus

bulgaricus trn mi trng MRS broth

Thi gian( gi)

Hnh 4.3: ng cong sinh trng ca L. acidophilus

Thi gian( gi)

Hnh 4.4: ng cong sinh trng ca L. bulgaricus

gi tip theo l pha cp s. V s lng t bo t cc i 18 gi. Pha cn bng din

Log( CFU/ ml)

Thi gian( gi)

Hnh 4.5: ng cong sinh trng ca L. acidophilus trong mi

Thi gian( gi)

Hnh 4.6: ng cong sinh trng ca L. bulgaricus trong mi trng