CHNG I

T VN
1. t vn
Ru vang l mt loi ru nh c ln men t dch p tri cy, l mt thc ung c
gi tr dinh dng cao, hng v thm ngon v c li cho sc khe con ngi khi dng mt
cch iu .
Trn th gii, nht l cc nc Chu u, ru vang c t lu i v c dng
rt ph bin. Nhng nc ta, sn phm ru vang vn cha c quan tm nhiu, qui m
sn xut ch yu di dng th cng, ln men t nhin cha c c s khoa hc cng nh
phng php hp l dn n nng sut cng nh cht lng sn phm cha cao.
Vi s pht trin ca khoa hc k thut, cng ngh sn xut ru bia cng c s
pht trin mnh m. Trong qu trnh sn xut ru truyn thng, ru nho c ln men t
nhin do nhiu chng nm men c sn trn tri nho. sn phm n nh v t cht lng
cao, cc nghin cu cn phi c tin hnh nhm phn lp v chn nhng ging nm men ti
u. Phn lp mt vi sinh vt l qu trnh tch ring tng vi sinh vt t mu vt hoc qun
th vi sinh vt ban u thu nhn ging dng thun khit. Sau cc ging ny c cho
ln men trong dch nho v kho st c tnh v tc pht trin trong iu kin khc nhau.
T trc n nay, nhiu nghin cu sn xut ru nho t nm men thun chng
Saccharomyces.cerevisiaeel loi nm men c bn ngoi th trng sn xut bnh m.
Do ti chn loi ny nghin cu v so snh tc ln men cng nh cht lng sn
phm ru thu c t nm men Saccharomyces.cerevisiaeev nm men phn lp ging
Saccharomyces sp t t nhin (trn v tri nho).
Vic nghin cu xem trong 2 ging nm men trn ging no c hot lc ln men mnh
nht, cht lng cao nht, c th to ra mt ru c cht lng tt hn, ang l mt nhu cu
thit thc nhm nng cao hiu sut len men cng nh cht lng sn phm.
2. Mc tiu nghin cu
T nhng iu trnh by trn, mc tiu ca ti l:
2.1 Kho st tc pht trin ca nm men, cn to thnh cng s thay i pH ca
dch qu nho theo thi gian khi tin hnh ln men dch qu bng ging nm men
Saccharomyces sp c phn lp t t nhin (trn v tri nho) v Saccharomyces cerevisiae.
2.2 So snh tc pht trin, cn to thnh i vi hai ging nm men kho st
trn. Qua kho st nh hng ca yu t pH ban u ca dch qu n tc ca tng
ging nm men v so snh.

CHNG II

LC KHO TI LIU
1. Nguyn liu
1.1 Nho
Trong sn xut ru vang, nho l mt nguyn liu truyn thng rt c a thch v c
gi tr dinh dng cao. Nho c trng nhiu cc nc Chu u nht l Php v M.
nc ta nho cng c trng mt s tnh min Trung v ang pht trin ngh trng nho
vi y trin vng pha trc.
Trn qu nho c rt nhiu loi nm men c mt mt cch t nhin l nhn t chnh
tham gia vo qu trnh sn xut ru vang nho ln men t nhin truyn thng.
Qu nho p c 85 95% dch qu. Nc nho ln men to ra mt loi ru vang c
gi tr dinh dng cao, hng v c trng, thm ngon v mu sc rt p.
Nho c thnh phn ha hc trung bnh nh sau:
-

Nc

: 70 80%

ng

: 10 25% (trong ch yu l glucose, fructose v saccharose)

Acid hu c : 0,5 1,7% (ch yu l acid malic v factoric)

Protein

: 0,1 0,9%

Pectin

: 0,1 0,3%

Khong

: 0,1 0,5%

Vitamin

: C, B1, B2, PP

Cc hp cht mu: mu chnh l anthocyanin.

Cc hp cht thm v mt s hp cht khc.

Nho l loi qu l tng nht ln men ch bin ru vang v:

-

Qu nho c cht lng, gi tr dinh dng cao, to hng v m , c trng.

Thnh phn ha hc trong dch nho rt thch hp cho nm men pht trin.

T l dch nc p c cao.

1.2. Nm men
Nm men l tn chung ch nhm nm men c cu to v sinh sn bng cch ny chi
v phn ct. Chng phn b rng ri khp ni. c bit chng c mt nhiu t trng nho
v cc ni trng hoa qu. Ngoi ra thy chng c mt trn tri cy chn, trong nhy hoa, trong
khng kh v c ni sn xut ru vang.
1.2.1 Hnh dng v kch thc
- Hnh dng t bo nm men
Nm men thng c hnh dng khc nhau, thng c hnh cu, hnh elip, hnh trng,
hnh bu dc v c hnh di. Mt s loi nm men c t bo hnh di ni vi nhau thnh
nhng dng si gi l khun ty (Mycelium) hay khun ty gi (Pseudo mycelium). Tuy nhin
hnh dng ca chng khng n nh, ph thuc vo tui ca nm men v iu kin nui cy.

- Kch thc t bo nm men

T bo nm men thng c kch thc rt ln gp t 5 10 ln t bo vi khun.
Kch thc trung bnh
- Chiu di: 9 10 m
-

Chiu rng: 2 7 m

Kch thc cng thay i, khng ng u cc loi khc nhau, cc la tui khc
nhau v iu kin nui cy khc nhau.
1.2.2. Cu to t bo nm men
T bo nm men cng nh nhiu loi t bo khc c cu to ch yu t cc phn c
bn nh sau:
-

Thnh t bo: cu to t nhiu thnh phn khc nhau. Trong ch yu l: glucan,

manan, protein, lipid v mt s thnh phn nh khc nh kitin, volutin,

Mng nguyn sinh cht: gm cc hp cht phc tp nh protein, phospholipit

enzyme permeaza

Cht nguyn sinh: thnh phn cu to ch yu l nc, protit, gluxit, lipit v cc

mui khong, enzyme v c cc c quan trong .

Nhn t bo

Nhng thnh phn c quan con khc: khng bo, ty lp th, riboxom,

1.2.3.S sinh sn ca nm men

Nm men c mt s hnh thc sinh sn sau:
- Sinh sn bng cch ny chi.
- Sinh sn bng cch phn i.
- Sinh sn bng bo t v s hnh thnh bo t.
+ Tip hp ng giao
+ Tip hp d giao
+ Sinh sn n tnh
1.2.4. Phn loi nm men
Ch yu c hai lp: nm men tht (Ascomyces) v lp nm men gi (Fungi imporfecti)
- Lp nm men tht (lp Ascomyces lp nm ti):
+ Phn ln nm men dng trong cng nghip thuc lp Ascomyces, a s thuc
ging Saccharomyces.
+ Ging Endomyces
+ Ging Schizosaccharomyces
- Lp nm men gi (Fungi imporfecti nm men bt ton)
+ Crytococus (toscula, tornlopsis)
+ Mycoderma
+ Eandida

+ Geotrichum ( c xp vo lp nm mc)
+ Rhodotorula
1.2.5. Cc qu trnh sinh l ca t bo nm men
* Sinh dng ca nm men
Cu to ca t bo nm men thay i khc nhau ty theo loi, tui v mi trng
sng, nhng nhn chung bao gm:
- Nc: 75 85%
- Cht kh: 15 25% . Trong cht khong chim 2 14% hm lng cht kh.
Bng I.1. Thnh phn ha hc ca nm men
Cc cht

Thnh phn (% cht kh)

Cacbon

49,8

CaO

12,4

Nitro

6,7

Hydro

3,54

P2O5

2,34

K2O

0,04

SO3

0,42

MgO

0,38

Fe2O3

0,035

SiO

0,09
Ngun: Lng, Nguyn c, 1996

Nm men cng nh cc sinh vt sng khc cn oxy, hydro, cacbon, nit, phospho, kali,
magi,
- Dinh dng Cacbon
Ngun Cacbon cung cp l cc loi ng khc nhau: saccarose, maltose, lactose,
glucose
H hp hiu kh:
C6H12O6 + 6O2

6CO2 + 6H2O + 674 cal

H hp k kh
C6H12O6

2CH3CH2OH + 2CO2 + 33 cal

- Dinh dng oxy, hydro: c cung cp cho t bo t nc ca mi trng nui cy

hay dch.
- Dinh dng Nit:
Nm men khng c men ngoi bo phn gii protid, nn khng th phn ct
albumin ca mi trng m phi cung cp nit dng ha tan, c th l m hu c hoc v
c. Dng hu c thng dng l acid amin, pepton, amid, ur. m v c l cc mui amon
kh nitrat, sulfat
- Cc vitamin v cht khong:
Cht khong c nh hng to ln n hot ng sng ca nm men
4

+ Phospho: c trong thnh phn nucleoprotein, polyphosphat ca nhiu enzyme ca

sn phm trung gian ca qu trnh ln men ru, chng to ra lin kt c nng lng ln.
+ Lu hunh: tham gia vo thnh phn mt s acid amin, albumin, vitamin v enzyme.
+ Magi: tham gia vo nhiu phn ng trung gian ca s ln men.
+ St: tham gia vo cc thnh phn enzyme, s h hp v cc qu trnh khc.
+ Kali: cha nhiu trong nm men, n thc y s pht trin ca nm men, tham gia
vo s ln men ru, to iu kin phc hi phosphorin ha ca acid pyruvic.
+ Mangan: ng vai tr tng t nh magi.
* C ch vn chuyn cc cht dinh dng vo trong t bo nm men
Nm men hon ton khng c c quan dinh dng ring bit, cc cht dinh dng m
n s dng ch yu c vn chuyn qua thnh t bo theo hai con ng c bn
- Thm thu b ng: trn thnh t bo nm men c nhng l nh, nhng l ny c tc
dng lm cho cht dinh dng vn chuyn vo trong t bo t mi trng bn ngoi nh p
sut thm thu, ngc li cht thi trong qu trnh trao i cng c thi ra theo con ng
ny.
- Hp thu ch ng: cc thnh phn dinh dng khng c kh nng xm nhp vo t
bo theo con ng th nht th lp tc c h permeaza hot ha. Permeaza l mt protid hot
ng, chng lin kt vi cht dinh dng to thnh hp cht v hp cht ny chui qua thnh t
bo trong, ti y chng li tch ra v permeaza li tip tc vn chuyn tip.
* Qu trnh sinh trng v pht trin
- S sinh trng
Trong qu trnh nui cy, trong iu kin dinh dng y , t bo nm men tng
nhanh v kch thc v ng thi sinh khi c tch ly nhiu.
- S pht trin
Cc nm men sinh sn bng phng php nhn i thng cho lng sinh khi rt ln
sau mt thi gian ngn. Trong trng hp sinh sn theo phng php ny th trong dch nui
cy s khng c t bo gi. V rng t bo c phn chia thnh hai c nh vy t bo lc no
cng trng thi ang pht trin. T bo ch gi khi mi trng thiu cht dinh dng v t
bo khng c kh nng sinh sn na.
Tuy nhin a s nm men sinh sn bng phng php ny chi nn hin tng pht
hin t bo gi rt r. Khi chi non tch khi t bo m sng c lp th ni tch trn t
bo m to thnh mt vt so. Vt so ny s khng c kh nng to ra chi mi. C nh vy
t bo m s chuyn thnh t bo gi theo thi gian.
xc nh s lng t bo nm men pht trin theo thi gian hin nay ngi ta dng
nhiu phng php khc nhau nh:
+ Xc nh s lng t bo bng phng php m trc tip trn knh hin vi hay gin
tip trn mt thch.
+ o c ca t bo trong dung dch nui cy trn c s xy dng mt th chun
ca mt t bo
Qu trnh sinh trng ca nm men trong dch ln men tnh c th chia lm 5 giai
on:

t
H1.1.Qu trnh sinh trng v pht trin ca nm men

Giai on tim pht: giai on ny t bo lm quen vi mi trng, sinh khi cha

tng nhiu.

Giai on logarit: y l giai on pht trin rt nhanh, sinh khi tng o t, km

theo s thay i mnh m ca dch ln men.

Giai on chm dn: tc sinh trng nm men gim dn, thnh phn dch ln
men cn li t, cc sn phm ln men c tch t nhiu.

Giai on n nh: s lng t bo nm men khng tng na, tc sinh sn bng

tc cht.

Giai on cht: tc cht tng nhanh, tc sinh sn rt t do s lng t bo

nm men gim dn.

1.2.6. Cc hnh thc h hp ca nm men

nm men h hp l qu trnh h hp kh phc tp, n xy ra theo hai chiu hng
khc nhau. V th ngi ta phn thnh 2 loi h hp : h hp hiu kh v h hp ym kh.
2. Khi qut v qu trnh ln men ru
Ln men l mt qu trnh trao i cht di tc dng ca cc enzyme tng ng gi l
cht xc tc sinh hc. Ty theo sn phm tch t sau qu trnh ln men m ngi ta chia lm
nhiu kiu ln men khc nhau. Tuy nhin c hai hnh thc ln men chnh l ln men ym kh
v ln men hiu kh.
Ln men ru l qu trnh ln men ym kh vi s c mt ca nm men, chng s
chuyn ha ng ln men thnh ethanol v CO2.
Qu trnh ln men ru chia lm hai thi k chnh:
- Thi k pht trin sinh khi: giai on ny vi s c mt ca oxy, t bo nm men
pht trin sinh khi.
- Thi k ln men chuyn ng thnh ru v CO2: giai on ny nm men hp th
cc cht dinh dng v s dng cc enzyme sn c ca mnh thc hin xc tc sinh hc trong
qu trnh trao i cht duy tr s sng, to thnh ru v CO2.

3. C ch ca qu trnh ln men
Glucose
Hexokinaza
Glucose 6 phosphat
Photphoglucoza isomeraza
Fructose 6 phosphat
PhosphoFructokinaza
Fructose 1,6 diphosphat
Aldolaza
Triophosphat izomeraza
Glyceraldehyd 3 phosphat

Dihydro aceton phosphat

Glyceraldehyd phosphatdehydrogenaza
Acid 1,3 diphosphoglyceric
Phosphoglyceratkinaza
Acid 3 phosphoglyceric
Phosphoglycerat-mutaza
Acid 2 phosphoglyceric
Enoiaza
Acid phosphoenolpyruvic
Pyruvat kinaza
Acid enol pyruvic
Acid pyruvic
Pyruvate decarboxylaza
Ethanal
Aldodeshydrogenaza
Ethanol
Hnh II.1. C ch phn hy ng trong t bo nm men

4. Khi qut v ru vang

Sn xut ru vang da trn c s v ha sinh xy ra trong qu trnh ln men cc loi
nc qu di tc dng ca enzyme ca nm men.
Trong nc qu (nho, mn, du, da, m) c cha ng glucose, fructose, cc cht
pectin, cc acid hu c (acid tartric, malic, succinic) v mui ca nhng acid ny, cc cht
mu, hp cht cha nit (protein, acid amin), vitamin cng nh cc mui khong

Trong qu trnh ln men ng trong dch qu c nm men s dng tng sinh

khi v tng hp mt s sn phm (ru, kh CO2 v glycerin, acid acetic, acid lactic, este
etylacetat). Cc alcol bc cao, aldehyd acetic c to thnh t cc acid amin. Cc cht
pectin b thy phn ko theo s to thnh mt lng nh metanol.
Ln men ru vang thng chia thnh cc giai on: ln men chnh nhit t 20
300C khong 10 ngy hoc di hn. cui giai on ln men chnh dch ln men trong dn v
protein v pectin lng xung. Ln men ph nhit t 15 180C. Khi lng cn hon ton,
dch trong th gn, lc xong s c ru vang c th ung c, nhng cha ngon, cn phi
tng tr nhit 4 100C ru vang hon thin hng v c trng. Thi gian tng tr
c th l vi thng, vi nm, thm ch hng chc hoc hng trm nm.
Ngi ta chia ru vang theo mu sc, theo hm lng ng c trong ru hoc theo
cn:
Theo mu sc: c vang trng, vang .
Theo lng ng cn li trong ru c: vang cht hay vang kh (ht ng) v vang
ngt (cn ng).
Ngoi ra cn c ru vang np CO2 hoc gi CO2 trong ln men.
Sn xut ru vang da trn c s bin i ha sinh xy ra trong qu trnh ln men cc
loi nc qu di tc dng ca h enzyme ca nm men. Hin nay, c hai phng php ln
men ru vang c bn: ln men t nhin v ln men nh cc chng nm men thun khit.
Phng php sn xut ru vang t chng thun khit c rt nhiu trin vng: thi
gian ln men nhanh, qu trnh ln men khng b dng gia chng, hm lng ng trong
dch qu c ln men trit , nng cn thu c trong vang cao hn ln men t nhin l
0.1 10, vang sng mu nhanh hn, c th cho hng v thanh khit hn.
5. H vi sinh vt trong ln men ru vang t nhin
H vi sinh vt trong ln men ru vang t nhin tng i phc tp v khng ng
nht trong cc giai on ca qu trnh ln men.
Trong nc nho ti c nhng nhm vi sinh vt khc nhau t mi trng xung quanh,
ch yu v qu, thn, cung v thit b. Phn ln trong phc h ny l nm mc (76 90%),
nm men (9 22%), s cn li chim t l thp l vi khun khng sinh bo t hoc c bo t,
x khun v Micobacter.
acid ca nc nho kh cao (pH = 2,7 3,8) l iu kin khng thun li cho cc vi
sinh vt pht trin.
mi trng c chua ln thng ch thch hp vi nm mc v nm men. Nm
men trong nc qu ti thng t hn nm mc, nhng nm men li c kh nng pht trin
v tng sinh khi nhanh trong iu kin thiu oxy hoc k kh, ng thi ln men tch t cn.
iu kin k kh v trong dung dch c cn lm c ch nm mc. Chnh trong iu kin ny
nm men cnh tranh v pht trin chim u th trong qu trnh ln men t nhin.
Trong cc loi nm men cng cnh tranh nhau ch c cc loi c kh nng ng ha
ng nhanh to cn cao mi dn dn chim u th giai on ln men chnh v ln men
ph.
Cc cng trnh nghin cu u xc nh rng, h nm men trong giai on u ln men
nc nho l kloeckera nm men c dng hnh chy chim phn ln v hot ng tch t
c 2 4 cn ri ngng hot ng v cht dn, sau l nm men ru vang thc th
8

(Saccharomyces cerevisiae v Saccharomyces oviformics) pht trin v ng vai tr ch yu

trong ln men chnh v ln men ph.
6. Mt s loi nm men thng gp trong sn xut ru vang
Sau y l mt s loi nm men thng gp trong nc qu c vai tr quan trng trong
ngh lm ru vang.
6.1. Saccharomyces cerevisiae
y l tn hin nay dng ph bin, trc y ngi ta gi l Saccharomyces
cerevisiae Meyer hay l S. ellipsoideus theo Lodder l Saccharomyces vini.
Nm men ny ph bin trong qu trnh ln men nc qu chim ti 80% trong tng s
Saccharomyces c trong nc qu khi ln men. Kh nng kt lng ca n ph thuc vo tng
ni: cc t bo dng bi hoc dng bng. Ngun dinh dng cacbon ca loi ny l ng,
cn v acid hu c, nhng tc nhn sinh trng l acid pantotinic, biotin, mezoinozit, thiamin
v piridoxin.
a s cc t bo ca loi ny hnh ovan c kch thc (3 8) x (5 12) m, sinh sn
theo li ny chi v to thnh bo t. Saccharomyces cerevisiae sinh ra enzyme invectara c
kh nng kh ng sacaroza thnh fructoza v glucoza, v vy trong ln men ta c th b
sung loi ng ny vo dung dch qu v hm lng ru c to thnh bnh thng i
vi nhiu ni ca men ny ch t c 8 10% so vi th tch.
giai on cui ln men Saccharomyces cerevisiae kt lng nhanh v lm trong dch
ru.
ni ca ging ny c c tnh ring v kh nng to cn, chu sunfit, tng hp cc
cu t bay hi v cc sn phm th cp to ra cho vang c mi v c trng ring bit.
Giai on cui cng ca qu trnh ln men cc t bo Saccharomyces cerevisiae
thng b gi, khng tip tc chuyn ng thnh cn v b cht rt nhanh.
6.2. Saccharomyces uvarum
Men ny c tch t nc nho, ru len men t nhin. V hnh thi n khng khc
vi cc loi khc. Kh nng sinh bo t kh mnh trn mi trng thch malt. Cc ni ca
loi ny c th ln men 12 130 cn trong dung dch nc nho. Mt vi ni c dng trong
sn xut ru vang.
6.3. Saccharomyces chevalieri
Theo Lodder l Saccharomyces chevalieri Guilliermond. Nm men ny c tch t
nc nho ln men t nhin, t ru vang non c gy men nc da hoc nc c.
Saccharomyces chevalieri thun chng ln men nc nho c th to 160 cn. N thng ln
vi Saccharomyces cerevisiae.
6.4. Saccharomyces oviformics
Theo Lodder l Sac. Beuanes saccardo. c tch ra t nc nho t ln men, nhng
loi nm men ny t hn so vi Sacch. vini. Ging thun chng pht trin tt trong nc nho
v cc loi nc qu khc, c kh nng chu c ng cao, cn cao, ln men kit ng v
to thnh ti 180 cn.
Cc yu t sinh trng ca loi ny ging nh Sacch. vini v c kh nng chu c
cn cao. Dng cc ni thun chng ca ging ny ln men dch qu c hm lng ng cao
ch vang kh cho kt qu tt.
9

C hnh dng ging nh Saccharomyces cerevisiae v c th to thnh 18% ru trong

qu trnh ln men, ging ny to thnh mng trn dch qu. S. oviformis ln men c
glucose, fructose, mantose, saccarose, maltose v 1/3 rafinose, khng ln men c lactose,
pentose. iu khc nhau c bn ca S. oviformis vi S. vini l: S. oviformis khng ln men
c galactose v men ni ln b mt dch ln men to thnh mng.
Hai ging sn xut ru vang ny (S. vini v S. oviformis) c nhiu ni c dng
trong sn xut.
6.5. Hanseniaspora apiculate Kloeckera apiculata
Kloeckera apiculata: kch thc tng i nh, c hnh ovan elip hoc hnh qu
chanh, t bo c mt u nh ngi ta thng gi l men hnh chy. Sinh sn bng ny chi,
rt ph bin v qu v nhim vo nc qu chim n 90% tng s men khi bt u ln
men. N c th ln men to thnh 6 70 cn, nhng to ra mt lot cc acid bay hi cng nh
cc este ca chng lm cho dch c mi tp v n cn km hm cc loi nm men chnh trong
ln men, K. apiculata nhy cm vi SO2.
Trong ngh lm ru vang ngi ta khng mong mun loi men ny pht trin, nu c
th ch cn c trong giai on u to c 3 40 cn.
7. Yu cu i vi chn nm men thun chng
Cc loi nm men thun khit dng nhiu trong sn xut ru vang thuc ging
Saccharomyces cerevisiae v Saccharomyces oviformis.
Cc chng nm men thun khit ny, c s khc nhau v tc sinh trng, khong
nhit thch hp ln men, kh nng to cn v chu cn, kh nng chu c pH thp
cng nh kh nng kt lng (to thnh dng bng hoc dng bi).
- Nhng yu cu i vi nm men ru vang l:
+ C hot lc ln men cao i vi nc qu
+ S dng ng cho ln men gn nh hon ton
+ Kt lng tt
+ Lm trong dch ru nhanh
+ Chu c ru cao v acid ca mi trng cng nh cc cht st trng
+ To cho ru hng v thm ngon tinh khit
8. Cc yu t nh hng ti nm men trong ln men ru vang
8.1. Oxy
Hu ht cc chng nm men trong ln men ru vang thuc ging Saccharomyces.
Chng l nhm vi sinh vt k kh ty tin. Khi trong mi trng lng oxy nm men phn
hy ng dng lm ngun nng lng v cu to t bo tng sinh khi.
Trng hp thiu oxy (k kh) nm men s dng phn oxy ha tan trong mi trng
sinh trng v ch yu l ln men.
Trong qu trnh ln men giai on u yu cu oxy cao nht nm men sinh sn, pht
trin tng sinh khi. Nu c giai on nhn ging th cng cn phi cung cp oxy bng cch
lc hoc sc kh.

10

8.2. Nhit
Nhit ln men c nh hng n i sng ca nm men, n qu trnh ln men v
cht lng ca sn phm.
Nhiu cng trnh nghin cu v sn xut ru vang xc nh c khong nhit
ln men ru vang trng thch hp l 15 300C, nu ln men nhng thang thp hn th
cng tt. Cn ln men ru vang (nc qu ln vi xc qu) phi chit xut cc cht thm
v polyphenol t v qu nn cn thang cao hn, thng l 250C.
Nhit ln men cao, thi gian ca qu trnh ln men ngn, cn c th thp, ng
st cn nhiu v hng v ca sn phm c khi khng tt.
8.3. Hm lng ng
Trong nc qu thng c hm lng ng khng u do vy ngi ta thng b
sung thm ng saccaroza. a s cc loi nm men hot ng bnh thng trong mi trng
ng di 20%. C mt s chng hot ng mi trng c ng cao hn. Khi nhn
ging thng dng mi trng c ng thp di 10%.
8.4. pH ca mi trng
Trong thc t ln men nhng dch qu chua thng c ru vang ngon. i vi
dch qu thng c pH t 2.8 3.8. Khong pH ny nm men vn hot ng c. Vng
pH ti thch ca nm men l 4 6.
Trong sn xut ru vang ngi ta thng chun b mi trng nc qu c pH
bng 3.0 3.5.
8.5. Ngun Nit
a s trong nc qu c cc hp cht nit cung cp cho nm men. Tuy nhin cng
c trng hp khng ngun nit do cn b sung thm ngun nit. Trong trng hp
ny ngi ta thng dng amon sulphat (NH4)2SO4. Cng c th dng men t phn cho thm
vo mi trng. Nu dch qu qu chua dng tartrat amon-kali hay amon hydroxy trung ha
bt acid.
i vi dch nhn ging hoc hot ha ging th hn hp cc ngun nit v cc cht
sinh trng rt c ngha.
Trong nc qu thng c cc cht khong i vi nhu cu ca nm men. V vy,
khng cn phi b sung thm cht khong. Tuy nhin trong nghin cu cng nh trong nhn
ging c th thm ngun phospho kali dng mui phosphat v magi dng mui sulfat.
Ngoi ra chng oxy ha nc qu, ngi ta c th thm ha cht vo nc qu sau
khi p v trc khi ln men. Cht dng rng ri l SO2 (anhydrit sunphur). SO2 l ha cht
c cho php dng trong sn xut ru vang hu ht trn th gii v c tc dng nhiu
mt: chng oxy ha, lm gim hoc tiu dit vi khun c hi. Ngun SO2 ph bin trong ru
vang l natri sunfit Na2SO3. Khng nn dng qu liu lng cho php s lm cho ru vang
c mi kh chu v dit mt s vi khun c ch.

11

CHNG III

PHNG TIN V PHNG PHP NGHIN CU

1. Phng tin th nghim
1.1. a im th nghim
Th nghim c tin hnh ti phng th nghim B mn Cng Ngh Thc Phm,
Khoa Nng Nghip & Sinh Hc ng Dng, Trng i Hc Cn Th.
1.2. Thi gian thc hin
Thi gian thc hin t 28 2 n 21 5 nm 2005
1.3. Thit b v dng c
Cc thit b v dng c cn thit cho cc th nghim nh:
- T cy v trng
- Knh hin vi
- Lam m
- Ni thanh trng
- My o pH
- Chit quang k
- Dng c phn lp nm men: a petri, ng nghim, n cn
- Cc dng c khc.
1.4. Ha cht s dng
- Cc ha cht thanh trng: NaHSO3
- Mi trng nui cy
- Nguyn liu: nho, ng

12

2. Phng php th nghim

2.1. S th nghim
Nho
(ra,p, lc...)

X l

To khun lc trn
a petri

Phn lp

Ging nm men hot

lc cao nht ging
Saccharomyces. spp

Nhn ging

Nho
X l
(ra, p, thanh trng)

Phi ch
(pH cc ch tiu kho
st)

Gi ging

Ging nm men cn kho

st s.spp (ging t nhin)

Acid citric

Ging nm men
Sacchromyces.Cerevisia

Dch ln men

Dch 1

Dch 2

Ln men

Ln men

Kho st tc pht trin

Kho st tc pht trin

* Ghi ch: Saccharomyces. sp t bo hnh trng, khun lc mu trng trong.

2.2. Chun b mi trng nui cy nm men

a. Nguyn tc ca vic ch to mi trng:
Da trn c s nhu cu v cc cht dinh dng v kh nng ng ha cc cht dinh
dng ca tng loi vi sinh vt. so snh c tnh v tc pht trin ca chng th cc
ging phi c nui tng sinh trong cng iu kin c tnh cht sinh l ging nhau.

13

 m bo s cn bng v p sut thm thu gia mi trng v t bo vi sinh vt

nn cn iu chnh t l v nng cc cht trong thnh phn mi trng.
m bo cc iu kin l ha cn thit cho cc hot ng trao i cht ca vi sinh vt.
b. S thc hin:

Bt mi trng

Cn

Bnh tam gic

Nc ct

Nu cch thy

Kh trng
(1210C, 15 pht

Bo qun v kim tra

mi trng
Hnh III.5. S chun b mi trng nui cy nm men.

c. Thuyt minh s :
Mi trng s dng nui cy nm men l mi trng Sabouraud, c cng thc ch to
nh sau:
Special peptone

10g/l

Glucose

40g

Dextrose

20g/l

Nc

1 lt

pH (250C)

5.6-6.0
Lng, Nguyn c,2002

- Cn mi trng nui cy v ha tan trong nc ct vi hm lng xc nh nh sau:

Bt mi trng

70g

Nc ct

1000ml

- Cho hn hp trn vo bnh tam gic v nu cch thy cc thnh phn trong mi
trng ha tan trong nc.
- Dng bng gn bt kn ming bnh tam gic.
- Ra sch cc a Petri, ng nghim, pipette. Sau dng giy bt kn thanh trng.
- Tit trng mi trng, dng c nhit 1210C trong thi gian 15 pht.
14

- Ly mi trng v dng c ra khi ni tit trng.

- Phn phi mi trng t bnh tam gic v dng c. i vi a petri lng mi
trng phn phi vo c dy khong 2mm. i vi ng nghim lng mi trng cho vo
bng 1/3 chiu cao ca ng.
- Qu trnh phn phi mi trng vo dng c cha phi tun th mt s nguyn tc c
bn sau:
+ Mi trng khi c phn phi phi trng thi lng.
+ Cc thao tc phn phi cn phi nhanh, gn, kho lo mi trng khng b dnh
ln ming hay thnh ca dng c cha v phi hon thnh trc khi mi trng ha rn.
- Bo qun v kim mi trng;
+ i vi mi trng cha s dng, cn c bo qun ch mt, nhit
thp(khong 20-25 0C), hn ch tc dng ca nh sng v khng mi trng b kh.
+ Trc khi s dng li kim tra v khun ca mi trng, ta thng t chng vo ch
m nhit 370C trong 42-48 gi, sau ly ra quan st, loi b cc ng c vi sinh vt pht
trin.
2.3. B tr th nghim
2.3.1. Th nghim 1: Phn lp nm men
a. Mc ch:
Trong dch ln men, vi sinh vt tn ti dng hn hp gm nhiu loi khc nhau (nm
men, nm mc, vi khun). Mun nghin cu nhng c tnh sinh l, sinh ha hoc s dng
mt loi no th cn phi a chng v dng thun khit.
Phn lp nm men l qu trnh tch ring cc chng nm men t qun th ban u v
a v dng thun khit. Nm men dng thun khit l ging nm men c to ra t mt t
bo ban u.

15

b. S thc hin:

Nho

X l

To khun lc trn
a Petri

Nc nho

Phi ch
(pH = 4 4.3 ; Brix = 22)

Thanh trng bng

NaHSO3 (122mg/l)

Tch rng v nui cy

vo ng nghim

Cy truyn

Nhn ging trong

ng nghim

Nhn ging trong

bnh tam gic
Hnh III.6. S qu trnh phn lp nm men

c. Thuyt minh s :
- Nho sau khi x l ra sch, dng dao ct i thnh hai ming.
- mt na tri trn b mt ca a petri c phn phi mi trng vo, lc nh
cho na qu nho chy u trn b mt a.
- Ly na tri nho ra.
- Lt p a v t vo t vi sinh nhit 37 0C trong 24 n 48 gi cho khun lc
xut hin.
- Xc nh cc chng nm men bng cch quan st;
+ Hnh dng, kch thc v mu sc ca khun lc (bng mt thng).
+ Quan st hnh dng v kch thc ca t bo nm men di knh hin vi.
- Chn ra ging nm men tiu biu em cy vo ng nghim l ging c khun lc
trng trong, t bo hnh trng ln.
Lu : Mi thao tc trn phi c thc hin trong iu kin v trng:
+ Tay v b mt tip xc trong t sy c kh trng bng cn.
+ Khng kh trong t c kh trng bng tia UV.

16

+ Dng c phi c kh trng trn ngn la n cn.

- Kim tra thun khit ca ging mi va phn lp bng cch kim tra vt cy, kim
tra thun chng ca cc khun lc v kim tra t bo nm men di knh hin vi.
- Cy truyn bo tn ging va phn lp: Tay tri cm hai ng nghim (mt ng
ging v mt ng mi trng), tay phi cm que cy v kh trng trn ngn n cn cho n
khi nng que cy. Sau , dng ngn ci v ngn tr xoay nh nt ng nghim ra . Tip
theo h nng kh trng khng kh ming hai ng nghim. i que cy va ngui, kho
lo a que cy tip xc vi khun lc trong ng nghim. K tip rt que cy ra, khng que
cy chm vo thnh ng nghim v a vo ng nghim cha mi trng, lt que cy trn
mt thch theo kiu hnh ch chi. Cui cng, rt que cy ra, kh trng li phn khng kh ni
ming hai ng nghim ri y np ng li v kh trng li que cy sau khi s dng xong.
- Nhn ging trong ng nghim v trong bnh tam gic pht trin sinh khi kho
st tc ln men ca ging nm men v gi ging.
2.3.1. Th nghim 2: Kho st tc ln men ca ging nm men c phn lp
ging Saccharomyces sp v ging Saccharomyces.cerevisiaee t men bnh m
* Mc ch
Kho st kh nng ln men, hot lc, ru hnh thnh pH khc nhau cng Brix
22% ca dch qu theo thi gian ca ging nm men Saccharomyces sp c phn lp ra v
ging Saccharomyces.cerevisiaee cc ch tiu.
- S th nghim
Nho
X l (ra, p)
Dch qu
PH= 4.0-4.
ng, acid Citric

Phi ch

pH = 3.7 4.5, Brix = 220Bx

Thanh trng (NaHSO3 122g/l)

S.sp
Ln men

nm men
(2% ging)
S.cerevisiae

A1
(B1, B2)

A2
(B1, B2)

A3
(B1, B2)

Phn tch
- Phng php thc hin:

17

Nho sau khi loi b cung, ra sch cho vo my p ly dch qu v xc qu. Tch
ring phn xc qu, ly dch trong mang i phi ch. Sau em i thanh trng bng NaHSO3
122mg/l, khi NaHSO3 phn hy hon ton (sau khong 30 pht), cho nm men vo. + Phi
ch: dng pH k v chit quang k iu chnh dch qu pH v Brix t yu cu, nu
cha t b sung ng v acid citric cho at yu cu.
+ B sung nm men vi hm lng 2% ging.
+ Khuy o u, sau phn tch mu ban u N0. Sau khi phn tch mu ban
u, dch qu em i phn phi vo mt s chai thy tinh vi hm lng 250ml/chai.
+ Dng bng gn y kn chai.
+ Quan st qu trnh ln men c sau 12h ly mu ra phn tch. Mi chai c ly em
i phn tch theo thi gian, iu ny khng gy nhim vo cc mu khc khi ly mu.
. pH: o bng pH k
. T trng .
. Hm lng CO2.
. cn.
. Brix: o bng chit quang k
. T bo nm men:
+ m bng knh hin vi
+ Cy trn a petri m khun lc

- B tr th nghim
Th nghim c tin hnh vi 2 nhn t
Nhn t A:Gi tr pH thay i 3 mc
A1: 3.7
A2: 4.1
A3: 4.5
Nhn t B:Ging nm men thay i 2 mc
Ging B1(Saccharomyces. sp)
Ging B2(Saccharomyce. cerevisiae)

Ch :
+
Song song vi vic phn lp nm men Saccharomyces.sp th ging nm men
Saccharomyces .cerevisiae cng c cy trn a v ng nghim tng t nh ging
Saccharomyces.sp .
+ Mc ch : to t bo nm men ging c trng thi sinh l tng ng vi t bo c
phn lp t t nhin. T ta c th kho st tc pht trin c thun li.

18

CHNG IV

KT QU V THO LUN
4.1 Th nghim 1:Phn lp nm men
Tham kho cc nghin cu cc ti trc v qua qu trnh phn lp chng ti chn
c ging nm men c trng nht, hot lc ln men cao nht t t nhin trn b mt tri nho
c dng t bo nm men hnh trng ln khun lc trng trong.
Phn lp nm men Saccharomyces. sp t nho thu nhn ging dng thun khit. Sau
qu trnh phn lp nhiu ln, ging nm men t nhin v nm men bnh m men
Saccharomyces.cerevisiaee c quan st di knh hin vi.

Hnh IV.1 Dng khun lc ca hai ging nm men va phn lp

Kt qu cho thy rng cc t bo nm men ca ging phn lp t nhin Saccharomyces.

sp tt v Saccharomyces. cerevisiaee c hnh dng tng t: hnh trng ln khun lc trng
trong.
4.2 Th nghim 2: So snh s pht trin ca 2 ging nm men

Hnh IV.2 Th nghim kho st tc ln men ca cc ging nm men

Sau cc ging thun khit ny c nui cy tng sinh cng iu kin v thi gian
to t bo mi c cng trng thi sinh l tng ng kho st.
19

4.2.1 nh hng ca pH ti tc phat trin, Brix, cn theo thi gian khi ln men
dch nho bng ging nm men saccharomyces.sp
10.00

SACCHA. SPP
pH=3.7
Ph=3,7

log10

9.00

SACCHA. SPP
pH=4,1
SACCHA. SPP
pH=4,5

8.00
7.00
6.00
5.00
0

24

48

72
96
thi gian

120

168

Hnh IV.3 nh hng ca pH n tc pht trin ca nm men theo thi gian ln men
(Phng php m khun lc trn da Petri)

SACCH. SPP

pH=3.7
Ph=3,7

Brix

20.00

SACCH. SPP
pH=4,1
SACCH. SPP
pH=4,5

15.00

10.00

5.00
0

24

48

72

96

120

168

thi gian

Hnh IV.4 nh hng ca pH n s gim Brix theo thi gian ln men

SACCH. SPP

pH=3.7
Ph=3,7

lng cn(%v)

12.00

SACCH. SPP
pH=4,1
SACCH. SPP
pH=4,5

8.00

4.00

0.00
0

24

48

72

96

120

168

thi gian

Hnh IV.5 nh hng ca pH n lng cn sinh ra theo thi gian ln men

20

11.00

SACCHA. SPP

log10

10.50

pH=3.7
Ph=3,7

10.00
9.50

SACCHA. SPP
pH=4,1
SACCHA. SPP
pH=4,5

9.00
8.50
8.00
7.50
7.00
0

24

48

72

96

120 168

thi gian
Hnh IV.6 nh hng ca pH n tc pht trin ca nm men theo thi gian ln men
(Phng php m t bo nm men di kinh hin vi)

Hnh IV.2 Th nghim kho st tc ln men ca cc ging nm men

(pp m khun lc trn a petri)
21

4.2.2 nh hnh ca pH ti tc phat trin, Brix, cn theo thi gian khi ln men
dch nho bng ging nm men saccharomyces.cerevisiae
10.00

S.CEREVISIA
pH=3,7
S.CEREVISIA
pH=4,1
S.CEREVISIA
pH=4,5

9.00
log10

8.00
7.00
6.00
5.00
0

24

48

72

96

120 168

thi gian

Brix

Hnh IV.7 nh hng ca pH n tc pht trin ca nm men theo thi gian ln men
(Phng php m khun lc trn da Petri)

23.00
21.00
19.00
17.00
15.00
13.00
11.00
9.00
7.00
5.00

S.CEREVISIA
pH=3,7
S.CEREVISIA
pH=4,1
S.CEREVISIA
pH=4,5

24

48

72 96
thi gian

120 168

Hnh IV.8 nh hng ca pH n s gim Brix theo thi gian ln men

14.00

S.CEREVISIA

l ng cn(%v)

12.00

pH=3.7
Ph=3,7

S.CEREVISIA
pH=4,1

10.00
8.00

S.CEREVISIA
pH=4,5

6.00
4.00
2.00
0.00
0

24

48 72 96
thi gian

120 168

Hnh IV.9 nh hng ca pH n lng cn sinh ra theo thi gian ln men

22

log10

11.00
10.50
10.00
9.50
9.00
8.50
8.00
7.50
7.00

S.CEREVISIA
pH=3,7
S.CEREVISIA
pH=4,1
S.CEREVISIA
pH=4,5

24

48 72 96
thi gian

120 168

Hnh IV.10 nh hng ca pH n tc pht trin ca nm men theo thi gian ln men
(Phng php m t bo nm men di kinh hin vi)

Vi phng php nui cy nm men trong mi trng Sabouraud, Hnh IV.3 biu din s
tng s lng t bo sng ca nm men. Trong khi hnh IV.6 l tng s t bo nm men sng
v cht khi im di knh hin vi. Do phng php im di knh hin vi khng phn
bit c t bo sng v cht nn s t bo tng mi.
T nhng th trn cho ta thy trong 24h u ca ca tin trnh ln men lng ng
gim t v s lng t bo nm men cn thp, trong khong thi gian ny s hnh thnh ru
rt t do t bo nm men tp trung pht trin sinh khi.
Sau 24h, tc pht trin ca t bo nm men rt mnh m (pht trin theo hm s m)
lm Brix gim mnh. ng thi lng ru sinh ra cng tng rt nhanh. Hnh IV.3 cho
thy, tc pht trin t bo t cc i trong khong thi gian t 48-72h, nng t bo nm
men cc i (Nmax) khong 1010 cfu/ml. Sau s t bo sng gim. iu ny c th gii thch
vi hai l do: lng ng cn t nn c s cnh mi trng sng rt cao gia hng triu t
bo nm men (1010 cfu/ml). L do th hai l lng ru trong mi trng sng rt cao nn c
ch s pht trin t bo nm men. Do s lng t bo sng gim xung sau 72h. ng thi
lng ng cng gim chm, nn lng ru tng chm.
Tht vy, hnh IV.5 cho thy lng cn tng nhanh t 48-72h, sau tng chm do hm
lng ng cn t, nhiu s t bo nm men cht. th biu din s thay i pH trong qu
trnh ln men cho thy pH t thp nht thoi em 48h, iu ny c th gii thch l khi ln
men, ngoi sn phm ru cn c sn phm ph l acid hu c. Sau 48 gi, pH tng ln l do
lc lng ng cn t mt s vi sinh vt c th s dng acid lm nng lng. Tht vy,
sau 96h lng ng cn rt thp v pH tng nhanh do lng acid ng s dng nhiu
thay cho ng cung cp nng lng cho vi sinh vt duy tr s sng.
T cc th cho thy tc pht trin v lng cn sinh ra ca ging nm men
Saccharomyces.sp t cao nht trong mi trng dch nho pH=3.7 so vi cc pH 4.1, 4.5.cn
vi ging saccharomyces.cerevisiaee th cao nht pH=4.1.

23

4.2.3 so snh s pht trin ca hai ging nm men saccharomyces.sp v

saccharomyces.cerevisiaee cc pH khc nhau
10.00
9.50

sacch.spp

log10

9.00
sacch.cerevisia

8.50
8.00
7.50
7.00
6.50
6.00
0

24

48

72

96

120

168

thi gian
Hnh IV.11 So snh tc pht trin ca Ssaccharomyces.sp v Saccharomyces.cerevisiaee pH=3.7
(Phng php m khun lc trn da Petri)

23.00
21.00

sacch.spp

19.00

sacch.cerevi
sia

Brix

17.00
15.00
13.00
11.00
9.00
7.00
5.00
0

24

48

72
96
thi gian

120

168

Hnh IV.12 So snh s gim Brix gia ging Saccharomyces.cerevisiaee v Saccharomyces.sp pH=3.7

14.00
sachh.spp

l ng cn (%v)

12.00
10.00

sacch.cer
evisia

8.00
6.00
4.00
2.00
0.00
0

24

48

72

96

120

168

thi gian
Hnh IV.13 So snh lng cn sinh ra gia ging sacc.sp v sacc.cerevisiaee pH=3.7

24

11.00

sacch.spp

10.50

sacch.cerevisia

log10

10.00
9.50
9.00
8.50
8.00
7.50
7.00
0

24

48

72

96

120

168

thi gian
Hnh IV.14 So snh tc pht trin ca saccharomyces.sp v saccharomyces.cerevisiaee pH=3.7
(Phng php m t bo nm men di kinh hin vi)

Bng VI..1 Bng thng k Brix, tc pht trin ca nm men cc phng php kho st khc nhau

Nhn t
kho st
Brix
s lng t
bo nm
men
(PP cy)
s lng t
bo nm
men
(PP m)

GING NM MEN
SACCH. SP
pH=4,1

pH=4,5

13.3a

13.7143a

920.443a

10508.7a

pH=3,7

pH=3,7

S.CEREVISIAEE
pH=4,1

pH=4,5

14.0714a

14.0714a

13.31a

14.0714a

903.586a

518.814a

661.429a

939.986a

903.586a

7975.71a

7502.71a

6750.0a

11317.4a

9761.14a

Bng VI.2 Nng ru sau khi kt thc qu trnh ln men

Nhn t
kho st
ru
cao nht

Saccharomyces.sp

Saccharomyces.cerevisiae

pH=3.7

pH=4.1

pH=4.5

pH=3.7

pH=4.1

pH=4.5

12.82

11.73

12.31

11.91

12.51

12.38

Qua qu trnh kho st tc pht trin ca cc ging nm men bng phng php m
cng nh phng php cy trn a petri chng ti thu c nhng kt qu sau:
Cc bng v hnh cho thy t bo ca hai ging nm men ny u pht trin chm trong
giai on u ca tin trnh ln men trc 24h, sau tc ca chng tng nhanh (trong
khong thi gian t 24-72h ) theo hm s m. Nm men t nhin biu hin c tc pht trin
nhanh hn v gim t bo nm men bnh m. iu ny c trnh bi t cc th trn: t bo
tng nhanh, lng ng gim nhanh, lng cn sinh ra nhanh v pH gim nhanh, ni chung
cc bin i din ra trc. Tuy nhin, kt qu thng k cho thy khng c s khc bit ngha

25

95% v tc pht trin, lng ng, lng cn sinh ra, pH gim trong dch ln men ca hai
ging nm men cc iu kin pH khc nhau.
4.2.3 nh gi cm quang sn phm ru to thnh
Bng IV.3 Tng hp kt qu nh gi cm quan
Cm
quan
vin

pH=3,7
Ln

1
2
1
2
2
1
3
2
1
4
2
1
5
2
1
6
2
1
7
2
1
8
2
1
9
2
1
10
2
1
11
2
1
12
2
1
13
2
1
14
2
1
15
2
Tng cu tr
li dng c hai
ln nh gi
1

Mu
sc
S
S

S
S

S
S

S
S

S
S

pH=4,1

Mi

S
S
S

S
S

S
S

S
S

S
S

S
S

S
S
S

Mu
sc
S
S
S

S
S

S
S

15

15

15

14

pH=4,5

Mi

S
S

S
S

S
S

S
S

S
S
S

S
S

S
S

S
S

S
S

Mu
sc

S
S

S
S

S
S
S

15

13

14

Mi

S
S

S
S

S
S

S
S

S
S

S
S

S
S

S
S

S
S

14

13

Kt qu:
pH=3,7
Ch tiu kho
st

pH=4,1

pH=4,5

Mu
sc

Mi

Mu
sc

Mi

Mu
sc

Mi

14

15

14

15

15

13

14

14

13

Ln
Tng cu
ng c 2 ln
nh gi

26

Bng IV.4 Tng hp ghi ch mc sai bit trong hai ln nh gi

Mc khc bit
Rt t
Hi nhiu
Nhiu
Rt nhiu

Kt qu nh gi
98
25
7
1

T kt qu nh gi cm quan trn cho thy sn phm ru vang nho c ln men t

hai ging nm men Saccharomyces. sp v Saccharomyces. cerevisiaee c mc khc bit
nhau l rt t.
T kt qu nh gi cm quan cho thy khng c s khc bit v mu sc cng nh mi,
v ca hai ging nm men cc pH khc nhau vi tin cy =99.9% . T y c th nhn
nh hai ging nm men c th chnh l mt ging.

27

Ghi ch: Bng thng k phn ph lc xc nh mc sai bit v tin cy

CHNG V
KT LUN V NGH
I.kt lun
Sau khi tin hnh kho st cc th nghim ca cc ging nm men rt ra mt s kt
lun sau.
Tc pht trin ca nm men c phn lp t t nhin Saccharomyces.sp v nm
men Saccharomyces cerevisiaee khc bit khng c ngha 95% trong cc iu kin pH 3.1,
4.1 v 4.5. Nng t bo ti a t c trong iu kin ln men l 1010 cfu/ml.
Thi gian ln men ca qu trnh ln men dch nho khi nm men c kch hot ln
mnh nht v vi nng ng 22%, 2% men ging chnh thc ch khong 07 ngy l kt
thc tin trnh ln men chnh. Nng ru ti a (12%) khng khc bit c ngha 95% khi
ln men nho t hai ging men trong dch nho c pH 3.1, 4.1 v 4.5 .
C th ging nm men c phn lp t t nhin Saccharomyces.sp chnh l ging
nm men Saccharomyces.cerevisiaee
C th s dng nm men phn lp t t nhin sn xut ru nho va t cht lng
cao, hiu qu khng km ging men mua.
II. ngh
V thi gian c hn ln vic nghin cu ch dng li vic phn lp v kho st ging
t t nhin vi ging Saccharomyces.cerevisiaee cc pH khc nhau. Trong qu trnh kho
st, kt qu cho thy hai ging khng c s khc nhau.
c th a ra nhng ch tiu tt hn cho phng php so snh c chnh xc hn cn:

Kho st nh hng ca Brix

Kho st nh hng ca nhit

Nghin cu v tin hnh nh danh ging nm men phn lp t t nhin

Saccharomyces.sp t nhin kho st xem c phi chnh l ging
Saccharomyces.cerevisiaee

28

PHN PH LC
Cc th bin tc pht trin ca cc ging nm men pH=4.1,4.5

I.

1.1 So snh s pht trin ca hai ging nm men Saccharomyces.sp v

Saccharomyces.cerevisiaee cc pH khc nhau
sacch.spp pH=4,1

10.00
9.50

sacch.cerevisiaepH=4
,1

log10

9.00
8.50
8.00
7.50
7.00
6.50
6.00
0

24

48

72

96

120

168

thi gian

Hnh I.1 So sanh tc pht trin ca Saccharomyces.sp v Saccharomyces.cerevisiaee pH=4.1

(Phng php m khun lc trn da Petri)

23.00
21.00

sacch.spp

19.00

sacch.cerevi
sia

Brix

17.00
15.00
13.00
11.00
9.00
7.00
5.00
0

24

48

72
96
thi gian

120

168

Hnh I.2 So snh s gim Brix gia gingSsaccharomyces.cerevisiaee v Saccharomyces.sp pH=4.1

29

14.00
12.00
lng cn(v%)

10.00
8.00
sacch.spp pH=4,1

6.00

sacch.cerevisiaepH=4,1

4.00
2.00
0.00
0

24

48

72

96

120

168

thi gian

Hnh I.3 So snh lng cn sinh ra gia ging Saccs.sp v Ssacc.cerevisiaee pH=4.1

11.00
10.50

log10

10.00
9.50
9.00

sacch.spp pH=4,1

8.50

sacch.cerevisiaepH=4,1

8.00
7.50
7.00
0

24

48

72

96

120

168

thi gian
Hnh I.4 So snh tc pht trin ca saccharomyces.sp v Saccharomyces.cerevisiaee pH=4.1
(Phng php m t bo nm men di kinh hin vi)
sacch.spp pH=4,5

10.00

sacch.cerevisiaepH=4
,5

9.50

log10

9.00
8.50
8.00
7.50
7.00
6.50
6.00
0

24

48

72
96
thi gian

120

168

Hnh I.5 So sanh tc pht trin ca Saccharomyces.sp v Saccharomyces.cerevisiaee pH=4.5

(Phng php m khun lc trn da Petri)
30

25.00

sacch.spp pH=4,5

Brix

20.00

sacch.cerevisiaepH=4,5

15.00
10.00
5.00
0.00
0

24

48

72

96

120

168

thi gian

Hnh I.6 So snh s gim Brix gia ging saccharomyces.cerevisiaee v saccharomyces.sp pH=4.5

14.00
lng cn(%v)

12.00
10.00
8.00
6.00

sacch.spp pH=4,5

4.00

sacch.cerevisiaepH=4
,5

2.00
0.00
0

24

48

72

96

120

168

thi gian
Hnh I.7 So snh lng cn sinh ra gia ging Saccs.sp v Sacc.cerevisiaee pH=4.5

11.00
10.50

log10

10.00
9.50
9.00
sacch.spp pH=4,5

8.50

sacch.cerevisiaepH=4,5

8.00
7.50
7.00
0

24

48

72

96

120

168

thi gian

Hnh I.8 So snh tc pht trin ca Saccharomyces.sp v Saccharomyces.cerevisiaee pH=4.5

(Phng php m t bo nm men di kinh hin vi)

31

I.2 S bin i c pHtheo thi gian ln men

S THAY I pH THEO THI GIAN LEN MEN

pH

4.60
SACCHA. SPP
Ph=3,7

4.40

SACCHA. SPP
pH=4,1

4.20

SACCHA. SPP
pH=4,5

4.00

S.CEREVISIA
pH=3,7

3.80

S.CEREVISIA
pH=4,1

3.60

S.CEREVISIA
pH=4,5

3.40
3.20
3.00
0

24

48

72

96

120

168

thi gian

II. PHNG PHP XC NH HM LNG RU

1.1 Nguyn tc
Xc nh hm lng ru da vo hm klng CO2 sinh ra.
xc nh ham lng ru c trong dch ln men da vo t trng ca dung dch trc khi bt
u ln men v khi kt thc qu trnh ln men.
Di tc dng ca t bo nm men, ng trong dch nho c chuyn thnh ru v CO2.
C6H12O6

2 C2H5OH

2 CO2

2 C2H5OH

2 CO2

1.2 cch tnh

C6H12O6

2*46.0688g

2*44.0098g

Da vo phng trnh phn ng trn c 46.0688g C2H5OH to thnh th c 44.0098g CO2 to

thnh theo v bay i. Hay ni cch khc 1.05g C2H5OH tng dng vi 1g CO2 .
Gi s t trng ca dch nho trc khi ln men l 1.06 v sau khi ln men l 1.02. nh vy trong
1 lt dung dch nho c 0.04 kg CO2 hnh thnh. Vy lng cn sinh ra l:
0.04*1.05=0.042 (kg/l)
M t trng ca dung dch sau khi ln l 1.02
32

Vy lng cn c trong dch ln men l:

0.042/1.02=4.1%
T trng ca cn l 0.79 kg/l
Vy % theo th tch l : 4.1/0.79=5.2
Ch : cn loi b CO2 trc khi lm th nghim trong phng php ny.
III.

PHNG PHP NH GI CM QUAN BNG PHNG PHP TAM GIC

II.1
II.2
II.3

Mc ch:So snh s khc bit v mt ch tiu no gia hai mu

Hi ng:10-20 thnh vin d c hun luyn.
S mu:3 (trong c hai mu ging nhau v mt mu khc).
II.4 Quy trnh:
Mi thnh vin nhn c ba mu ghi m s, c bit trc trong c hai mu ging
nhau. Yu cu cc thnh vin phi ch ra mu no khc hn so vi mu kia. Cc thnh vin
cng c th c yu cu nhn xt mc khc bit cng nh so snh cht lng gia mu
khc v hai mu kia. Nhng yu cu nu c i vi sn phm cung c th c ngh. Ni
dung ca bng nh gi theo phng php tam gic c th tm tt nh bng di y.
II.5 p dng:
Chng minh hoc khng chng minh s khc bit gia hai ging, hai ln giao nguyn liu
hoc sn phm sn xut ra nhng ngy khc nhau, nhng ca khc nhau. Phng php ny cn
c dng hun luyn thnh vin hi ng.

1.

2.

BNG NH GIA CM QUAN

H v tn.....................................................................................................................
Tn sn phm..............................................................................................................
Ch tiu nh gi ........................................................................................................
Cc mu c b tr v nh gi theo phng php tam gic.
Ni dung:
Ch ra mu khc bit trong 3 mu ghi m s
356...............................................................................................................
514...............................................................................................................
487...............................................................................................................
Nhn xt mc khc bit
Rt t.............................................................................................................
Hi nhiu ....................................................................................................
Nhiu ...........................................................................................................
Rt nhiu......................................................................................................
3. So snh cht lng
Mu khc tt hn..........................................................................................
Hai mu ging nhau tt hn.........................................................................
4. Nhng yu cu khc ...........................................................................................
.....................................................................................................................
.....................................................................................................................

33

Bng tra tin cy cng nh mc ngha trong nh gi cm quan(pp tam gic)

Triangle test, difference analysis
Number
of tasters

number of correct answers

necessary to establish
level of significance
*
**
***
5%
1%
0.1%

Number
of tasters

number of correct answers

necessary to establish
level of significance
*
**
***
5%
1%
0.1%

7
8
9
10

5
6
6
7

6
7
7
8

7
8
8
9

57
58
59
60

27
27
27
28

29
29
30
30

31
32
32
33

11

61

28

30

33

12

10

62

28

31

33

13

10

63

29

31

34

14

10

11

64

29

32

34

15

10

12

65

30

32

35

16

10

11

12

66

30

32

35

17

10

11

13

67

30

33

36

18

10

12

13

68

31

33

36

19

11

12

14

69

31

34

36

20

11

13

14

70

32

34

37

21

12

13

15

71

32

34

37

22

12

14

15

72

32

35

38

23

13

14

16

73

33

35

38

24

13

14

16

74

33

36

39

25

13

15

17

75

34

36

39

26

14

15

17

76

34

36

39

27

14

16

18

77

34

37

40

28

15

16

18

78

35

37

40

29

15

17

19

79

35

38

41

30

16

17

19

80

35

38

41

31

16

18

19

81

36

38

41

32

16

18

20

82

36

39

42

33

17

19

20

83

37

39

42

34

17

19

21

84

37

40

43

35

18

19

21

85

37

40

43

36

18

20

22

86

38

40

44

37

18

20

22

87

38

41

44

38

19

21

23

88

39

41

44

39

19

21

23

89

39

42

45

40

20

22

24

90

39

42

45

41

20

22

24

91

40

42

46

42

21

22

25

92

40

43

46

43

21

23

25

93

40

43

46

44

21

23

25

94

41

44

47

45

22

24

26

95

41

44

47

46

22

24

26

96

42

44

48

47

23

25

27

97

42

45

48

48

23

25

27

98

42

45

49

49

23

25

28

99

43

46

49

50

24

26

28

100

43

46

49

51

24

26

29

200

80

84

89

52
53
54

25
25
25

27
27
27

29
29
30

300
400
500

117
152
188

122
158
194

127
165
202

55

26

28

30

1000

363

372

383

56

26

28

31

2000

709

722

737

34

IV.

CC BNG THNG K
a. Brix

Multiple Range Tests for brix by Gio_pH

-------------------------------------------------------------------------------Method: 95,0 percent LSD
Gio_pH
Count LS Mean
Homogeneous Groups
-------------------------------------------------------------------------------13
7
13,3
X
241
7
13,3
X
141
7
13,7143
X
245
7
14,0714
X
145
7
14,0714
X
23
7
14,0714
X
-------------------------------------------------------------------------------Contrast
Difference
+/- Limits
-------------------------------------------------------------------------------13 - 23
-0,771429
6,58026
13 - 141
-0,414286
6,58026
13 - 145
-0,771429
6,58026
13 - 241
0 ,0
6,58026
13 - 245
-0,771429
6,58026
23 - 141
0,357143
6,58026
23 - 145
0,0
6,58026
23 - 241
0,771429
6,58026
23 - 245
0 ,0
6,58026
141 - 145
-0,357143
6,58026
141 - 241
0,414286
6,58026
141 - 245
-0,357143
6,58026
145 - 241
0,771429
6,58026
145 - 245
0,0
6,58026
241 - 245
-0,771429
6,58026
--------------------------------------------------------------------------------

35

b. T trng

Analysis of Variance for Ti Trong - Type III Sums of Squares

-------------------------------------------------------------------------------Source
Sum of Squares Df Mean Square F-Ratio P-Value
-------------------------------------------------------------------------------MAIN EFFECTS
A:Gio_pH
0.00169878 5 0.000339756
0.23 0.9483
RESIDUAL
0.0538539 36 0.00149594
-------------------------------------------------------------------------------TOTAL (CORRECTED)
0.0555527 41
-------------------------------------------------------------------------------Multiple Range Tests for Ti Trong by Gio_pH
-------------------------------------------------------------------------------Method: 95.0 percent LSD
Gio_pH
Count LS Mean
Homogeneous Groups
-------------------------------------------------------------------------------241
7
1.03159
X
13
7
1.03406
X
141
7
1.03619
X
23
7
1.04246
X
145
7
1.04296
X
245
7
1.0504
X
-------------------------------------------------------------------------------Contrast
Difference
+/- Limits
-------------------------------------------------------------------------------13 - 23
-0.0084
0.0419288
13 - 141
-0.00212857
0.0419288
13 - 145
-0.0089
0.0419288
13 - 241
0.00247143
0.0419288
13 - 245
-0.0163429
0.0419288
23 - 141
0.00627143
0.0419288
23 - 145
-0.0005
0.0419288
23 - 241
0.0108714
0.0419288
23 - 245
-0.00794286
0.0419288
141 - 145
-0.00677143
0.0419288
141 - 241
0.0046
0.0419288
141 - 245
-0.0142143
0.0419288
145 - 241
0.0113714
0.0419288
145 - 245
-0.00744286
0.0419288
241 - 245
-0.0188143
0.0419288
--------------------------------------------------------------------------------

36

c. cn sinh ra theo thi gian

Analysis of Variance for con _sra - Type III Sums of Squares
-------------------------------------------------------------------------------Source
Sum of Squares Df Mean Square F-Ratio P-Value
-------------------------------------------------------------------------------MAIN EFFECTS
A:Giong_pH
0.0022334 5 0.00044668
0.18 0.9699
RESIDUAL
0.0914554 36 0.00254043
-------------------------------------------------------------------------------TOTAL (CORRECTED)
0.0936888 41
Multiple Range Tests for con _sra by Gio_pH
-------------------------------------------------------------------------------Method: 95.0 percent LSD
Gio_pH
Count LS Mean
Homogeneous Groups
-------------------------------------------------------------------------------245
7
0.0499143
X
145
7
0.0596143
X
23
7
0.0598571
X
141
7
0.0649143
X
13
7
0.0708143
X
241
7
0.0709
X
-------------------------------------------------------------------------------Contrast
Difference
+/- Limits
-------------------------------------------------------------------------------13 - 23
0.0109571
0.0546397
13 - 141
0.0059
0.0546397
13 - 145
0.0112
0.0546397
13 - 241
-0.0000857143
0.0546397
13 - 245
0.0209
0.0546397
23 - 141
-0.00505714
0.0546397
23 - 145
0.000242857
0.0546397
23 - 241
-0.0110429
0.0546397
23 - 245
0.00994286
0.0546397
141 - 145
0.0053
0.0546397
141 - 241
-0.00598571
0.0546397
141 - 245
0.015
0.0546397
145 - 241
-0.0112857
0.0546397
145 - 245
0.0097
0.0546397
241 - 245
0.0209857
0.0546397
--------------------------------------------------------------------------------

37

d. S lng t bo phng php cy trn a petri m khun lc

Analysis of Variance for toc_do_ppcay - Type III Sums of Squares
-------------------------------------------------------------------------------Source
Sum of Squares Df Mean Square F-Ratio P-Value
-------------------------------------------------------------------------------MAIN EFFECTS
A:Gio_pH
1.01293E6 5
202586.0
0.26 0.9344
RESIDUAL
2.85835E7 36
793987.0
-------------------------------------------------------------------------------TOTAL (CORRECTED)
2.95965E7 41
Multiple Range Tests for toc_do_ppcay by Gio_pH
-------------------------------------------------------------------------------Method: 95.0 percent LSD
Gio_pH
Count LS Mean
Homogeneous Groups
-------------------------------------------------------------------------------145
7
518.814
X
23
7
661.429
X
13
7
737.171
X
141
7
903.586
X
241
7
920.443
X
245
7
939.986
X
-------------------------------------------------------------------------------Contrast
Difference
+/- Limits
-------------------------------------------------------------------------------13 - 23
75.7429
965.966
13 - 141
-166.414
965.966
13 - 145
218.357
965.966
13 - 241
-183.271
965.966
13 - 245
-202.814
965.966
23 - 141
-242.157
965.966
23 - 145
142.614
965.966
23 - 241
-259.014
965.966
23 - 245
-278.557
965.966
141 - 145
384.771
965.966
141 - 241
-16.8571
965.966
141 - 245
-36.4
965.966
145 - 241
-401.629
965.966
145 - 245
-421.171
965.966
241 - 245
-19.5429
965.966
--------------------------------------------------------------------------------

38

e. S lng t bo phng php m di knh hin vi

Analysis of Variance for toc_do_pp_dem - Type III Sums of Squares
-------------------------------------------------------------------------------Source
Sum of Squares Df Mean Square F-Ratio P-Value
-------------------------------------------------------------------------------MAIN EFFECTS
A:Gio_pH
1.1602E8 5
2.3204E7
0.21 0.9574
RESIDUAL
4.03513E9 36 1.12087E8
-------------------------------------------------------------------------------TOTAL (CORRECTED)
4.15115E9 41
Multiple Range Tests for toc_do_pp_dem by Gio_pH
-------------------------------------------------------------------------------Method: 95.0 percent LSD
Gio_pH
Count LS Mean
Homogeneous Groups
-------------------------------------------------------------------------------23
7
6750.0
X
145
7
7502.57
X
141
7
7975.71
X
245
7
9761.14
X
13
7
10508.7
X
241
7
11317.4
X
-------------------------------------------------------------------------------Contrast
Difference
+/- Limits
-------------------------------------------------------------------------------13 - 23
3758.71
11477.1
13 - 141
2533.0
11477.1
13 - 145
3006.14
11477.1
13 - 241
-808.714
11477.1
13 - 245
747.571
11477.1
23 - 141
-1225.71
11477.1
23 - 145
-752.571
11477.1
23 - 241
-4567.43
11477.1
23 - 245
-3011.14
11477.1
141 - 145
473.143
11477.1
141 - 241
-3341.71
11477.1
141 - 245
-1785.43
11477.1
145 - 241
-3814.86
11477.1
145 - 245
-2258.57
11477.1
241 - 245
1556.29
11477.1
--------------------------------------------------------------------------------

39

TI LIU THAM KHO

1. Hu, V Cng,1993, CH BIN RU VANG TRI CY TRONG GIA NH.
2. Tr, H Quang,1999, VI SINH THC PHM.
3. Lng, Nguyn c,1996, Phc, Nguyn Hu, 1996, CNG NGH VI SINH
VT(tp 1.2.3) Trng i Hc Bch Khoa TP.HCM
4. Lng, Nguyn c, 2002, TH NGHIM CNG NGH SINH HC, Tp 2, Trng
i Hc Bch Khoa TP.HCM
5. Phm, Lng c, 1998, CNG NGH VI SINH VT. NXB Nng Nghip.
6. Tr Nhan Minh, Teshome Edae Jiru, Naznin Sultana, Michael Wawire, 2001,
FERMENTATION PROCESSING.

40