Professional Documents
Culture Documents
Thng 08 nm 2009
B GIO DC V O TO
TRNG I HC NNG LM THNH PH H CH MINH
KHOA CNG NGH SINH HC
******************
Hng dn khoa hc
TS. V VN
CN. TH TUYN
Thng 08 nm 2009
LI CM N
Trc tin em xin by t lng bit n su sc n thy V Vn v c
Th Tuyn ht lng hng dn, gip v to mi iu kin thun li cho em trong
sut qu trnh thc hin ti.
Xin by t lng bit n chn thnh i vi qu thy c trong Khoa Cng Ngh
Sinh Hc trng i hc Nng Lm thnh ph H Ch Minh cng nh qu thy c l
gio vin thnh ging ti cc khoa, trng, vin nghin cu tn tnh ging dy em
trong thi gian hc tp ti trng.
Xin chn thnh cm n n ban lnh o Vin sinh hc Nhit i, qu c, ch,
anh, ch v cc bn sinh vin ang cng tc v lm vic ti vin cng ngh sinh hc
nhit i, v i bo v gip em trong qu trnh thc hin ti.
Xin gi li cm n n cc bn lp DH05SH, i hc chnh quy kha 2005
-2009 quan tm v ng vin ti trong thi gian hc tp v thc hin ti.
Cui cng, con xin by t lng bit n su sc n ba, m v nhng ngi thn
trong gia nh ng vin, gip v chia s cng vi con trong thi gian qua.
iii
TM TT
Enzyme amylase l mt enzyme c th tm thy rt nhiu sinh vt. Hin nay,
ezyme ny c ng dng rng ri trong nhiu lnh vc nh nng nghip, cng nghip
v y hc. Mc tiu ti l nghin cu kh nng to enzyme amylase ca nm
Aspergillus niger v Mucor v nhng yu t tc ng n kh nng to amylase ca
chng. ti thu c mt s kt qu v s tc ng ca cc yu t mi trng n
kh nng sinh tng hp enzyme amylase t nm Aspergillus niger v Mucor.
Aspergillus niger khi sinh trng v tng hp enzyme amylase trn mi trng
bn rn vi c cht l cm go b sung thm tru v b u nnh chu tc ng ca rt
nhiu yu t mi trng nh t l c cht, pH mi trng, thi gian nui cy, nhit ,
m, nng dinh dng v t l ging. ti nghin cu s tc ng ca cc
yu t mi trng nu trn u tin l mc ring l tng yu t, sau l s
tng tc qua li ca nhng yu t c nh hng mnh n s tng hp enzyme. Kt
qu thu c cho thy vi nhng mi trng c t l Cm go: tru: b u nnh
(2:1:1), pH 5, thi gian nui cy l 28 gi, nhit 350C, m 55%, 2 ln nng
dinh dng, t l ging 0,8*108 bt/g l ti u cho Aspergillus niger sinh tng hp
amylase.
Kh nng tng hp enzyme amylase t Mucor khi sinh trng trn mi trng
bn rn vi c cht l cm go b sung thm tru v b u nnh chu tc ng ca rt
nhiu yu t mi trng nh t l c cht, pH mi trng, thi gian nui cy, nhit ,
m, nng dinh dng v t l ging. ti nghin cu s tc ng ca cc
yu t mi trng nu trn u tin l mc ring l tng yu t, sau l s
tng tc qua li ca nhng yu t c nh hng mnh n s tng hp enzyme. Kt
qu thu c cho thy vi nhng mi trng c t l Cm go: tru: b u nnh
(2:1:1), pH 5, thi gian nui cy l 32 gi, nhit 300C, m 55%, 3 ln nng
dinh dng, t l ging 1,6*108 bt/g l ti u cho Mucor sinh tng hp amylase.
iv
SUMMARY
Enzyme amylase can be found in many living organisms. Nowadays, this is
applied in many domains, such as agriculture, industry and medicine. The purpose of
the survey is to find out Aspergillus niger and Mucors ability to produce enzyme
amylase, and some factors can affect this ability. Finally, the survey has some
following results
The first, for Aspergillus niger, they are born and grow up on semi-solid
environments including rice bran, rice rind, and soya residuum. Growing up and
biosynthesized enzyme ability of A. niger depend on environment factors, such as ratio
of rice bran to rice rind and to soya residuum, time, humidity, nutritious concentration,
and ratio of spore. The survey finds out affect of environment factors in individual of
each and compose of some factors, which have stronger affect. Results showed with
environments have ratio of rice bran to rice rind and to soya residuum is 2:1:1; after 28
hours at 350C with 55% humidity, double nutritious concentration and ratio of 0,8*108
spore to one g environment is the most suitable for Aspergillus niger to biosynthesize
enzyme amylase on semi-solid fermatable environment.
The second, for Mucor, they are born and grow up on semi-solid environments
including rice bran, rice rind, and soya residuum. Growing up and biosynthesized
enzyme ability of A. niger depend on environment factors, such as ratio of rice bran to
rice rind and to soya residuum, time, humidity, nutritious concentration, and ratio of
spore. The survey finds out affect of environment factors in individual of each and
compose of some factors, which have stronger affect. Results showed with
environments have ratio of rice bran to rice rind and to soya residuum is 2:1:1, after 32
hours at 300C with 55% humidity, treble nutritious concentration and ratio of 1,6*108
spore to one g environment is the most suitable for Mucor to biosynthesize enzyme
amylase on semi-solid fermatable environments.
MC LC
Trang
LI CM N..............................................................................................................iii
TM TT....................................................................................................................iv
MC LC....................................................................................................................vi
DANH SCH CC CH VIT TT.........................................................................xii
Hnh 2.1 Cu trc phn t ca amylose Error: Reference source not found...............xiii
th 4.14 Hot tnh v hm lng theo cc nng ging ca Mucor Error:
Reference source not found........................................................................................xiv
DANH SCH CC BNG.........................................................................................xv
Chng 1.......................................................................................................................1
M U.......................................................................................................................1
1.1. t vn ...............................................................................................................1
Chng 2.......................................................................................................................3
TNG QUAN TI LIU..............................................................................................3
2.1. Gii thiu s lc v enzyme.................................................................................3
2.1.1. Khi nim chung v enzyme................................................................................3
2.1.2. Tnh cht u vit ca enzyme..............................................................................3
2.2. Tng quan v enzyme amylase...............................................................................4
2.2.1. Amylase...............................................................................................................4
2.2.2. Phn loi..............................................................................................................4
2.2.3. c tnh v c ch tc dng ca 3 loi amylase thng dng................................4
2.2.3.1. -amylase..........................................................................................................4
2.2.3.2. -amylase..........................................................................................................5
2.2.3.3. -amylase (hay gluoamylase hoc -1,4 glucan-glucanhydrolase)....................6
2.2.4.1. Thc vt............................................................................................................6
2.2.4.2. Vi sinh vt.........................................................................................................7
2.2.5. ng dng.............................................................................................................7
2.2.5.1. ng dng trong cng ngh thc phm..............................................................7
2.2.5.2. Trong cng nghip dt, giy..............................................................................8
vi
Chng 3.....................................................................................................................21
VT LIU V PHNG PHP NGHIN CU......................................................21
3.1. Thi gian v a im th nghim.........................................................................21
3.2. Vt liu th nghim................................................................................................21
3.2.1. i tng th nghim.........................................................................................21
3.2.2. C cht...............................................................................................................21
3.3. Phng php nghin cu.......................................................................................21
3.3.1. Phng php cy truyn.....................................................................................21
3.3.2. Phng php nhn ging cp 2 (gi ging trn mi trng la)........................22
3.3.3. Phng php m t hnh thi A. niger v Mucor...............................................23
3.3.3.1. Quan st hnh thi i th ...............................................................................23
3.3.4. Xc nh trc tip s lng bo t nm si bng bung m hng cu.............23
3.3.6. Phng php ln men bn rn thu nhn enzyme amylase.............................25
3.3.7. Phng php xc nh hm lng amylase........................................................26
Phng php ny da trn s thay i bc sng hp th cc i ca thuc th
Coomassie Brilliant Blue khi to phc vi protein. Trong dung dch mang tnh acid,
khi cha kt hp vi protein th thuc nhum c bc sng hp th cc i 465 nm,
khi kt hp vi protein th thuc nhum hp th cc i bc sng 595 nm. hp
th bc sng 595 nm c lin h mt cch trc tip vi nng protein. c bit
nu c mt cc acid amin nhn thm s bt mu mnh hn. Phng php ny c
nhy cao cho php pht hin ti vi g protein/ml, d thc hin v tit kim thi gian.
.....................................................................................................................................26
Bng 3.1 Cc bc tin hnh to dung dch albumin chun........................................27
3.3.8. Phng php xc nh hot tnh amylase...........................................................28
Enzyme amylase l enzyme thy phn tinh bt. N phn ct amylose v amylopectin
ca tinh bt thnh cc loi ng maltose, glucose,Hot tnh ca enzyme amylase
c xc nh da trn s thay i mu sc ca phc hp tinh bt iod trc v sau
khi thy phn. Mt quang ca phc hp tinh bt iod s c o bc sng 595
nm trn my o quang ph...........................................................................................28
Ha cht cn thit cho th nghim ny bao gm dung dch Lugol, dung dch m Naacetate 50 mM pH 5, dung dch h tinh bt 1%. Dung dch ny c pha bng cch
viii
cn 1 g tinh bt tan cho vo berche ng 100 ml dung dch m, un cch thy cho si
trong 5 pht, khuy u cho tan...................................................................................28
3.3.9. Phng php quy hoch thc nghim...............................................................29
3.3.10. Phng php b tr v x l s liu th nghim................................................29
Chng 4.....................................................................................................................30
KT QU V THO LUN.....................................................................................30
4.1. Kh nng sinh tng hp amylase ca nm mc Aspergillus niger trn mi trng
ln men bn rn
.....................................................................................30
ix
..................................................42
KT LUN V NGH..........................................................................................63
5.1. Kt lun.................................................................................................................63
5.2. ngh..................................................................................................................63
TI LIU THAM KHO............................................................................................64
xi
Aspergillus niger
B u nnh
bt
bo t
Cm go
CT
Canh trng
UI
One-Way ANOVA
PGA
Pt
phng trnh
Tru
TY
UV
Ultraviolet
xii
found
Hnh 4.2 Hnh nh i th v vi th ca Mucor. Error: Reference source not found
th 4.1 Hot tnh v hm lng theo t l c cht ca A. niger.
Error: Reference
not found
th 4.3 Hot tnh v hm lng theo thi gian ca A. niger Error: Reference source
not found
th 4.4 Hot tnh v hm lng theo nhit ca A. nigerError: Reference source
not found
th 4.5 Hot tnh v hm lng theo m ca A. niger.
not found
th 4.6 Hot tnh v hm lng theo nng dinh dng ca A. niger. Error:
Reference source not found
th 4.7 Hot tnh v hm lng theo t l ging ca A. niger.
Error: Reference
Error: Reference
xiv
Bng 2.1 Thnh phn dinh dng ca cm go Error: Reference source not found
Bng 3.1 Cc bc tin hnh to dung dch albumin chun
not found
Bng 4.1 M ha cc bin s cc yu t nh hng n A. niger
Error: Reference
Error:
Error: Reference
not found
Bng 4.6 Hot lc amylase ca Mucor theo thc nghim v pt hi qui
Reference source not found
xv
Error:
Chng 1
M U
1.1. t vn
Enzyme l mt loi protein c bit c sinh vt tng hp nn v tham gia cc
phn ng sinh hc, l thnh phn khng th thiu trong mi t bo sinh vt. Chng
ng vai tr quyt nh mi quan h gia c th sng v mi trng sng.
Enzyme amylase c vai tr rt quan trng trong nhiu ngnh cng nghip v
cha tr bnh, nh cng nghip thc phm, dt, ty ra, cha bnh km tiu ha
ngi v trong ngnh ch bin thc n gia sc
Cui th k 19 v sut th k 20, ngnh cng nghip sn xut enzyme rt pht
trin, ngy cng thin v xu hng sn xut enzyme da vo hot ng sng ca vi
sinh vt v nhiu li ch thit thc. Cng ngh sn xut enzyme v ang mang li
nhng ngun li ln cho nhiu quc gia trn th gii.
Hin nay, doanh thu t vic sn xut enzyme ln n hn 1,7 t USD, trong
gp phn khng nh l li nhun thu c t enzyme amylase. Th trng enzyme
cng ngh s cn tip tc gia tng trong tng lai n. nc ta, nhu cu s dng cc
ch phm enzyme trong c amylase trong ch bin thc phm, thc n gia sc v
trong cc ngnh cng nghip khc ngy cng nhiu. V vy, hng nm, nc ta phi
nhp khu lng ln nhng ngun enzyme ny.
Ngy nay, vic tn dng ph liu trong cng, nng nghip lm ngun carbon
sn xut amylase bi vi sinh vt bng phng php ln men chm v ln men bn rn
rt c quan tm nc ta v nhiu nc trn th gii. Vit Nam l mt nc nhit
i c nn nng nghip lu i v a dng, ang trn pht trin. V vy lng ph
phm nng nghip thi ra ngy cng gia tng, nh cm go, b u nnh l ngun
cung cp carbon thch hp cm ng nm mc sinh tng hp amylase bng phng
php ln men bm rn. Mc khc, y l c cht kh r tin v n nh c tim nng
s dng sn xut amylase qui m ln.
Hin nay, vin cng ngh sinh hc nhit i c mt s chng nm mc c kh
nng sinh tng hp enzyme amylase kh cao, trong c chng Aspergillus niger v
Mucor. Tuy nhin, thu c enzyme amylase c hiu sut cao ch cn chng vi sinh
vt c kh nng to ezyme amylase cao l cha . Cc vi sinh vt cn phi c
nui trn mi trng c cht thch hp c kh nng cm ng kch thch chng sinh
amylase. Mi trng nui cy cn c b sung thm nhng nng dinh dng v
cc vi sinh vt phi c nui nhit thch hp. Cui cng, thi gian thu nhn dch
chit enzyme t canh trng nui cy cng s nh hng rt ln n hm lng v
hot tnh amylase thu c.
Trn c s , chng ti chn ti Kho st mt s yu t nh hng n
kh nng sinh tng hp enzyme amylase t nm mc Aspergillus niger v Mucor trn
mi trng ln men bn rn nhm xy dng iu kin mi trng ti u cho hai
chng nghin cu to enzyme nhiu v c hot tnh cao cng nh iu kin thu
nhn enzyme tt nht. ti c thc hin ti phng cc cht c hot tnh sinh
hc - vin Sinh Hc Nhit i, thnh ph H Ch Minh.
1.2. Yu cu
- Xc nh cc gi tr ti u ca cc yu t mi trng nh hng n
kh nng sinh tng hp enzyme amylase ca hai chng th nghim.
- Xc nh mi tng quan gia cc yu t mi trng c tc ng mnh
n kh nng tng hp amylase ca hai chng th nghim.
1.3. Ni dung
- Kho st kh nng tng hp amylase ca hai chng th nghim.
- Kho st s nh hng ca cc yu t nh t l c cht, m, nhit
nui cy, thi gian nui cy, pH mi trng ban u, nng dinh dng,
v t l ging n kh nng sinh tng hp amylase ca hai chng th nghim.
- Ti u ha cc yu t trn bng phng php quy hoch thc nghim
nhm tm ra iu kin nui cy v thnh phn mi trng ti u cho hai chng
trn sinh ra lng amylase cao.
Chng 2
TNG QUAN TI LIU
2.1. Gii thiu s lc v enzyme
2.1.1. Khi nim chung v enzyme
Enzyme l mt cht xc tc sinh hc ch c to ra trong t bo sinh vt, v
ng vai tr quan trng khng th thiu c trong qu trnh trao i cht ca c th
sinh vt. Tt c cc phn ng chuyn ha cc cht trong c th sinh vt u c s tham
gia xc tc ca enzyme. Enzyme khng ch tham gia xc tc cc phn ng trong t bo
m ngay c ngoi t bo ca vi sinh vt. iu ny c ngha quan trng trong vic
ng dng enzyme vo nhiu lnh vc khc nhau ca cng nghip v i sng.
Enzyme c bn cht l protein c kh nng xc tc c hiu cho cc phn ng
ha hc v lm cht xc tc sinh hc. Di tc dng ca enzyme cc phn ng ha
sinh trong c th sinh vt xy ra rt nhanh, khng cn nhng iu kin v nhit , p
sut, nng cc cht mt cch qu khc nghit. Tn enzyme c bt ngun t ch
Hy Lp c ngha l cht trong con men. Tt c qu trnh bin i ha sinh u xy ra
di tc dng ca h men.
Nu c th thiu enzyme th cc qu trnh chuyn ha ha sinh s b nh ch,
sinh vt khng th sng, sinh trng, pht trin v sinh sn bnh thng. S sng ca
sinh vt s khng tn ti.
Ngy nay, enzyme cn ng vai tr quan trng trong cc hot ng sn xut
cng nghip v ngy cng c nhiu nhng nghin cu v ng dng enzyme vo trong
cc lnh vc khoa hc, cng nghip, kinh t quc dn, vn vn.
2.1.2. Tnh cht u vit ca enzyme
Th nht, enzyme c th thu nhn c t nhiu ngun khc nhau nh ng
vt, thc vt, v vi sinh vt, v hu nh cc enzyme khng c tnh cht c hi.
Th hai, chng c th hot ng xc tc phn ng trong iu kin nhit bnh
thng, p sut thng, pH acid yu, kim yu hay trung tnh.
Th ba, tnh c hiu ca enzyme cao hn hn cc xc tc khc, v ch c tc
dng ln nhng c cht nht nh v theo mt kiu phn ng nht nh.
100% glucose.
-amylase kiu
Asp.niger
Tinh bt hay oligosaccharide -------------> 80 85% glucose + oligosaccharide.
2.2.4. Ngun thu nhn
2.2.4.1. Thc vt
T lu con ngi bit cch s dng amylase t ht ny mm s dng
trong ngnh ch bin thc phm (mch nha), nc gii kht (bia), hoc sn sut ch
c bit l i vi ngnh cng nghip bia ca nc ta do hin nay vic gieo trng i
mch trong nc cha mang li nhiu kt qu, nn nc ta vn phi nhp malt t nc
ngoi vi gi thnh cao. V vy nu s dng enzyme amylase t vi sinh vt s gp
phn lm gim gi nguyn liu v gim s ph thuc vo vic nhp khu malt.
Trong sn xut bnh m s dng enzyme amylase c tc dng rt ngn qu
trnh ln men, tng nhanh th tch bnh m, lm tng mi thm ca bnh, mu sc ca
bnh p hn v tng xp ca bnh. Trong sn xut bnh ngt, nu thm ch phm
amylase vo bt s xy ra qu trnh thy phn bt thnh ng, do gim lng
nguyn liu tiu tn cho sn xut.
Trong sn xut mt, tinh bt, maltose, glucose nhu cu s dng enzyme
amylase cng ngy cng cao. Ban u cc sn phm mt, tinh bt thng c sn
xut theo phng php thy phn tinh bt bng acid hoc malt. Ngy ny, qui trnh
ny c thc hin nh ch phm amylase. Ch phm enzyme amylase ng dng rt
hiu qu trong sn xut tinh bt, glucose, v cc loi mt tinh bt. Khi thy phn tinh
bt bng acid khng cho php sn xut cc loi ng mt theo mun, bi v khi
phn t tinh bt b thy phn v trt t v khng th iu chnh c t lng ca cc
cu t ca dch thy phn. Phm cht ca ng mt, tnh cht ha l ca chng li
ph thuc vo t lng ca cc cu t chnh ny. Khi thy phn tinh bt bng ch
phm enzyme s khng ging nh thy phn bng acid, khng xy ra s thy phn cc
hp cht ln trong tinh bt nn lm dch thy phn tt hn, d dng tinh ch hn.
2.2.5.2. Trong cng nghip dt, giy
Trong cng nghip dt, amylase c s dng r h vi, ty lp h bt trn
vi lm vi mn, mm, d ty trng v d bt mu khi nhum. X l vi bng phng
php ezyme tt hn rt nhiu so vi vic x l bng cc cht ha hc. X l vi bng
enzyme khng ch nhanh m cn gi c bn, dai, khng hi vi, mao dn
tt v m bo v sinh. Do vy tng c nng sut lao ng.
2.2.5.3. Trong y hc
Amylase c dng phi hp vi cc coenzyme A, cytocrom C, carbonxydase
ch thuc iu tr cc bnh tim mch, bnh thn kinh, phi hp vi enzyme thy
phn cha bnh thiu enzyme tiu ha. Ngoi ra ch phm enzyme amylase dng
iu ch mi trng nui vi sinh vt phc v cho cng tc nghin cu khoa hc
cng nh chn tr bnh.
8
10
H
Hnh 2.2 Cu trc phn t ca amylopectin. (Wikipedia.com.).
C trng lng phn t khong vi triu dalton, c cu to t khong 600
n 6000 phn t D-glucose, ni vi nhau bi lin kt -1,4 glucoside v lin kt -1,6
glucoside to thnh mch c nhiu nhnh.
Amylopectin cho mu tm vi iod, ha tan trong nc nng cho dung dch
c nht cao. Thy phn tinh bt nh nhng vi amylase, hay acid long thi gian
ngn s to thnh dextrin, mt t maltose, glucose. Thy phn thi gian lu hn vi
acid hoc enzyme vi nhit cao hn s to nhiu maltose v glucose hn.
2.2.6.2. Glycogen
11
: Moniliales.
: Monilaceace.
Ging: Aspergillus.
Nhm: Aspergillus niger.
A. niger thuc loi nm si a bo. u si nm sinh sn phnh to ra to thnh
t bo hnh chai v cc chui nh bo t. Cung ca nh bo t n bo, t bo hnh
chai v chui nh bo t to u trn u si nm sinh sn nh nhng tia nc i t
xa vo vi ti. Cung sinh bo t c sinh ra t khun ty kh sinh, c chiu di t 1
n 3 mm, c vch t bo trn bng v trong sut. nh cung phnh ra c dng hnh
cu c gi l bng, c ng knh t 50 75 m, sinh ra hai lp th bnh. Lp th
nht hnh tam gic cn ngc, lp th hai hnh chai. Bo t hnh cu, dng bo t nh
c ng knh 4 5 m, c mu nu en, vch bo t nh c dng x x.
12
: Mucorales
: Mucoraceace
Ging: Mucor
Mucor l ging ln nht trong b Mucorales. Chng bao gm cc loi: M.
mucedo, M. cremois, M. rouxii, M. hiemalis, M. silvaticus, M. subtilis, Mucor l
mt loi nm pht trin rng ri, thng tm thy trong t, phn bn, tri cy, bnh
m, v nhng thc phm giu tinh bt khc. Chng pht trin tt trong iu kin hiu
kh. iu kin ym kh Mucor. sppr phn tch thnh nhng si nm nh hnh trn
hay hnh bu dc v ny chi, v th ngi ta hay gi loi ny l men ca nm mc.
Chng c kh nng s dng mt cch hiu kh nhiu ngun carbon, ln men
carbonhydrate, c th s dng ammoniac hay nit hu c cho qu trnh sinh trng, v
cn c th pht trin trn mt thang nhit rng. V th chng c mt trn nhiu loi
mi trng, bao gm nhiu loi thc n khc nhau.
13
tng hp amylase khc. Theo mc tiu th mui, mi trng s chuyn dch mnh
v pha tng hp glucoamylase v c ch tng hp -amylase.
S to amylase cc i thng thy cc nng nit cao (0,25 0,4 %). Khi
lm mi trng nui cy nm mc to amylase thng dng NaNO 3 l tt nht. Cng
c th b sung nit hu c (gelatin, casein, nc chit ng) nm sinh trng tt,
nhng li khng c tc dng kch thch to amylase. Thm nc chit mm mch hay
nc chit ng cng nh cc amino acid ring r s lm tng sinh tng hp -amylase
ngoi bo, song hiu qu khng bng NaNO3 (Nguyn c Lng, 2004).
Tnh cn bng ca mi trng dinh dng v carbon v nit c ngha ln i
vi sinh tng hp sinh khi vi sinh vt v s to thnh amylase. Ch khi no trong mi
trng c lng carbon v nit cn thit th vi sinh vt mi c kh nng tch ly
amylase cc ln. S thiu ht cu t ny khng th b p bng s tha cu t kia.
Thng thng, t l carbon gp t 1 n 40 ln hm lng nit trong mi trng l
thch hp nht. Trong mi trng Zapeck th t l ny l 18 : 1.
2.4.2.3. Cc nguyn t khong
Cc nguyn t khong a lng v vi lng c nh hng ln ti sinh trng
v tng hp cc enzyme amylase ca vi sinh vt. V cc nguyn t khong ny cn
c b sung vo mi trng theo t l thch hp kch thch s sinh tng hp
amylase. Nu mi trng thiu hoc tha khong u c nh hng n s to enzyme
ca cc vi sinh vt.
Mg2+ c nh hng n bn nhit. Thiu MgSO 4 s c nh hng xu n s
tng hp enzyme amylase ca nm si. Khi s tng hp -amylase s b c ch
hon ton, cn lng -amylase s b gim hng chc ln ( Fenikxova, Muxaeva,
1967). Nng ti u khong 0,05%. Tuy nhin, mui ny khng nh hng n s
to oligo-1,6-glucosidase (Silova, 1967).
Phospho cn tng hp cc hp phn quan trng ca sinh cht v nhiu
coenzyme, ng thi phosphoril ha glucide trong qu trnh oxy ha sinh hc.
Phospho nh hng trc tip ti sinh sn ca nm si cng nh cc vi sinh vt khc, v
vy cng nh hng n s sinh tng hp enzyme ca chng. Nhiu nh nghin cu
cho bit nu thm phospho hu c (nh dng fitin) vo mi trng s lm tng s tng
hp amylase ln 2 3 ln.
15
Khi lng/100g
316 KJ
21g
4g
21g
28 g
0,9 g
13,3 g
4,9 mg
4,1 mg
57 mg
20 mg
(http://www.nutritiondata.com/).
Theo bng trn th cm c lng dinh dng rt cao vi lng cht bo cha
bo ho cao, vitamin nhm E, nhm B, phylate, km, can xi, kali u rt cao, ngoi ra
trong cm cn c chc cht bo omega 3 kh cao. Theo nhiu nghiu cu cho thy
65% cht dinh dng ca go tp trung cm, thnh phn ca cm c nhiu loi
vitamin v cht bo tt, li cn i v c nhiu x d tiu nn c th xem l ngun
lng thc rt tt cho c con ngi.
Tuy nhin cm thng c cho cc loi gia sc v thy sn n ch khng cho
ngi. Nguyn nhn l do cm c mt s enzyme (cht men) ni ti hot ng rt
mnh s oxy ha cc nhm bo cha no ca cm rt nhanh ch vi gi sau khi ch bin
to mi hi kh chu. Thm vo do cng ngh xay xt go cha cao, li t c u
17
18
19
20
Chng 3
VT LIU V PHNG PHP NGHIN CU
3.1. Thi gian v a im th nghim
Th nghim c tin hnh t thng 2 n thng 7 nm 2009, ti phong cac cht
co hoat tinh sinh hc Vin sinh hoc nhit i.
3.2. Vt liu th nghim
3.2.1. i tng th nghim
Nm mc Aspergillus niger va Mucor do Vin Sinh Hc Nhit i cung cp
c nui cy trong ng nghim thch nghing trn mi trng PGA nhit
thng l 300C trong thi gian 2 - 3 ngy, sau o em bo qun nhit 40C.
3.2.2. C cht
C cht s dng trong ti bao gm lua, cm go, tru v b u nnh. Lua,
cm go, tru c mua ch Th c, cm phi tng i xp d thng kh, khng
c mi i. B u nnh c cung cp bi Vin Sinh Hc Nhit i. Cc c cht s
c phi trn vi nhau trong mi trng nui cy.
3.2.3. Hoa cht va thit bi
3.2.3.1. Hoa cht
Ha cht pha thuc th Lugol, ha cht pha mi trng Czapeck, mi trng
Zapeck, ha cht pha mi trng PGA, mi trng dinh dng Mandel, ha cht dng
xc nh hm lng protein, bao gm dung dch Abumin chun v thuc th
Bradford, ha cht dng chnh pH cho mi trng dinh dng (HCl 0,1M, NaOH
0,1M), cn 700, 900, tinh bt tan v dung dch m Na-acetate 50 mM pH 5.
3.2.3.2. Thit bi
Cc thit b gm c cn phn tch, b n nhit, phng m hng cu, knh hin
vi quang hc Olympus, ni hp Tommy-SS-325, my xay sinh t, my khuy t, t
lnh, my o pH, my o quang ph UV-Vis cng cc dng c phng th nghim nh
que cy, pipet, pipetman, ng nghim, ng ong, becher, erlen, vn vn.
3.3. Phng php nghin cu
3.3.1. Phng php cy truyn
Do c hai loi nm trn u sinh sn bng bo t nn ta s dng phng php
gi ging trn mi trng thch nghing, bng cch cy theo ng dch dc. Khi
21
thy lng bo t mc nhiu, em cc erlen cho vo tr trong t lnh nhit 5 90C dng cho cc th nghim sau.
3.3.3. Phng php m t hnh thi A. niger v Mucor
3.3.3.1. Quan st hnh thi i th
Dng que cy mc (v trng) gt nh bo t nh ca nm si, chuyn sang a
petri cha mi trng PGA v trng v ngui, cm u mc xung mt thch
thnh 3 im. Cc ng tc trn c thc hin trong t cy. Sau a petri nhit
phng trong khong thi gian 2 ngy. Quan st khun lc ca nm si (hnh dng
v mu sc).
3.3.3.2. Quan st hnh thi vi th
Lm phng m quan st hnh thi vi th. Lt giy thm y petri (ng
knh 12 cm) ri t vo ming lame, gi giy v em kh trng. Nu chy mi trng
PGA (v trng) rt chy dc theo na tm lame v ch ng li, lm m giy thm
bng nc ct v trng (va t t giy). Cy mt t bo t thnh mt hoc hai
im trn phn thch ca ming lame trong phng m, nui nhit phng. Sau 2
n 3 ngy ly ming lame ra khi hp petri v quan st. Quan st di knh hin vi
( phng i 40 v 100 ln) hnh dng vi th ca hai chng th nghim.
u tin dng que cy hnh mc cu mt t h si nm dn mng trn phin
knh. Tip theo cho vi git cn 95% mt lc, dng giy thm lau kh sau cho
vi git NaOH 10% y l knh ln, em soi. Hoc cho vi git cn 600 c vt
amoniac, thm kh ri cho thm vi git glycerin long ln tm lame, y l knh ln,
em soi. Mc ch ca cc tc dng trn l tng cng kh nng thm nc ca si
nm, ui bt kh trong si nm, lm si nm n to ra d quan st.
3.3.4. Xc nh trc tip s lng bo t nm si bng bung m hng cu
Bung m hng cu dng m vi sinh vt c kch thc ln (nm men, bo
t nm mc). Ngi ta thng dng 2 loi bung m hng cu; bung m Thomas
v bung m Goriep. Nguyn tc cu to ca 2 loi bung m ny u ging nhau.
Bung m u l 1 phin knh hnh ch nht, chia thnh 3 khong ngang. Khong
gia chia thnh 2 khong nh. Trn mi khong nh ny c k mt li m, gm rt
nhiu vung. Mi vung c din tch l 1/25 mm 2 li c chia thnh cc nh
(thng l 16 ), mi nh c din tch l 1/400 mm2 v chiu cao l 1/10.
Nh vy V nh = 1/400 mm2 x 0,1 mm = 1/4000 mm3 hay 1/4000000 ml.
23
a*4000*103*n
b
24
F=
= 40 (ml)
100
Nh vy: trong 10g c cht ta b sung thm 40ml nc ct s c m 50%.
Vi :
A
25
26
1
0
0,0
1,0
2
2
10
0,1
0,9
2
3
20
0,2
0,8
2
4
30
0,3
0,7
2
5
40
0,4
0,6
2
6
50
0,5
0,5
2
b*m*V
w
(mg/g)
Vi:
b : nng protein trong mu suy ra t ng chun (g/ml).
m : h s pha long.
V: tng s ml dch chit thu c.
w: tng khi lng canh trng thu c.
27
(A0-An)*100*V*n
(UI/g CT)
A0*30*w
Nu tnh hot lc enzyme amylase trong 1ml dch enzyme:
HT =
(A0-An)*100*n
(UI/ml)
A0*30
Trong :
A0 : l mt quang ca mu i chng bc sng 595 nm.
An : l mt quang ca mu enzyme bc sng 595 nm.
n
: h s pha long
28
HT chung
(UI/mg protein)
HL protein
3.3.9. Phng php quy hoch thc nghim
Phng php quy hoch thc nghim khng ch cho php nghin cu tc ng
ng thi ca nhiu yu t ln qu trnh m cn cho php nh gi mt cch nh
lng mc nh hng ca tng yu t .
Sau khi thc hin cc th nghim kho st s nh hng ca tng yu t, ta rt
ra c thnh phn mi trng v cc iu kin nui cy c s c gi l mi trng
c s cho hai chng trn sinh trng v to thnh enzyme amylase.
T mi trng c s ta tin hnh ti u ho cc yu t trn bng cch tin hnh
cc th nghim theo k hoch thc hin y cc yu t (gi tt l TY). C s ca
vic qui hoch thc nghim theo yu t 2n l vic th hin tt c cc t hp c th c
gia cc yu t nghin cu. Mi yu t u c kim tra ng thi v khng ph
thuc ln nhau hai mc l trn (+) v di (-). Trung tm ca th nghim l mi
trng c s xc nh c (Nguyn Cnh, 1993).
3.3.10. Phng php b tr v x l s liu th nghim
Tt c cc th nghim c b tr theo kiu hon ton ngu nhin vi 3 ln lp
li. S liu thu c ca cc nghim thc s c phn tch bin lng n yu t
(One-way ANOVA) nh gi s khc bit ca chng. y l phng php phn
tch s liu th nghim da trn nhng bin ng gy ra cc sai s trong th nghim.
Mc ch ca ANOVA l xc nh mc sai khc gia cc trung bnh nghim thc
ca ch tiu nghin cu l do nghim thc hay ch mang yu t ngu nhin. Ngoi ra,
ANOVA cn cho php so snh nhiu s trung bnh ng thi vi nhau c kt lun
chung v s khc bit. Ni cch khc, nu qua ANOVA m xc nh c gia cc
trung bnh nghim thc khc nhau c ngha th chng t rng s sai khc l do
cc nghim thc th nghim gy ra.
29
Chng 4
KT QU V THO LUN
4.1. Kh nng sinh tng hp amylase ca nm mc Aspergillus niger trn mi
trng ln men bn rn
4.1.1. Nm mc A .niger
Bo t A. niger t ng nghim thch nghing c cy sang a Petri v ming
lame trong phng m cha mi trng PGA theo phng php m t mc 3.3.3,
nui nhit phng 2 - 3 ngy. Kt qu quan st i th v vi th c trnh by
hnh 4.1 v 4.2.
(b)
(a)
30
th 4.1 Biu din hot tnh amylase v hm lng protein t canh trng nui cy
A. niger vi cc t l c cht khc nhau.
Hot tnh amylase v hm lng protein cao nht (358,552 UI/g CT v 41,484
mg/g) mi trng c t l cm: tru: b u nnh l 2:1:1. Hm lng v hot tnh
amylase mi trng c t l cm: tru: b u nnh l 1:2:1 cng kh cao. Mi
trng c t l cm: tru: b u nnh l 1:1:2 cho hm lng v hot tnh amylase
thp. iu ny cho thy vic tng tinh bt (c trong cm go) trong trng hp ny l
c cht ca enzyme c tc ng n vic kch thch vi sinh vt sinh tng hp enzyme
tng ng. Kt qu th nghim cng ch ra rng vic tng cng lng b u nnh
lm gim ng k hm lng v hot tnh ca amylase. C th do trong b u nnh
cha nhiu lipid, l c cht cho vic tng hp enzyme lipase nn c ch s tng hp
enzyme amylase. Tuy nhin, vic b sung thm b u nnh vi t l thp vo mi
trng s cung cp thm ngun Nit cho vi sinh vt.
Nng carbon v cc ngun carbon khc nhau cng c nh hng ln n s
to thnh cc enzyme ring bit ca h enzyme amylase khi nui nm mc. Trong ,
m bo c hot lc -amylase cao cn c t nht 6% tinh bt, cn mun c hot
31
32
th 4.2 Biu din hot tnh amylase v hm lng protein t mi trng nui cy
A. niger vi cc pH mi trng ban u khc nhau.
pH c nh hng rt mnh n sinh tng hp amylse ca A. niger. Khong pH
ti u cho loi nm mc ny sinh tng hp amylase l t 4 - 6, trong ti u nht l
ti pH 5. Ti pH ny, A. niger tng hp lng amylase nhiu nht v c hm lng
cao nht (354,333 UI/g CT v 41,961 mg/g). pH nm ngoi khong ti u ny, A.
niger tng hp rt t amylase v hot tnh enzyme cng rt thp.
Khi nui cy bng phng php b mt, pH mi trng t b thay i do mi
trng c dung lng m cao v hm m thp. V vy, pH ban u c ngha rt ln
n s pht trin ca nm mc v s to enzyme. Mi trng c pH trong khong t
4,5 5,0 s to iu kin chn lc cho nm mc pht trin do a s nm mc sinh
trng tt v to enzyme trong mi trng acid yu. V cng pH ny, nm mc to
h amylase tt nht. V th, canh trng nui cy c pH 5, lng enzyme v hot
tnh amylse thu c l cao nht. iu ny l ph hp vi cc nghin cu trc y.
pH cng c nh hng n vic to enzyme ring bit ca h enzyme amylase
ca nm si. Trong mi trng acid, qu trnh to enzyme s chuyn dch mnh v
pha tng hp tch cc glucoamylase v c ch s tng hp -amylase. pH thch hp
cho vic to glucoamylase namwg trong khong t 3 n 3,5, trong khi pH thch hp
cho vic to thnh -amylase nm trong khong pH 5.
33
th 4.3 Biu din hot tnh enzyme v hm lng protein t canh trng nui
cy A. niger vi cc mc thi gian thu nhn khc nhau.
Khong thi gian A. niger to enzyme amylase mnh m l sau 24 32 gi
nui cy. Hot tnh v hm lng enzyme cao nht t c sau 28 gi nui cy.
Trong khong thi gian t 8 n trc 24 gi nui cy, lng enzyme tng rt chm.
Hot tnh v hm lng enzyme tng mnh sau 24 n 28 gi nui cy. Sau 28 gi
nui cy, hot tnh enzyme gim nhanh nhng hm lng protein gim chm. Sau 36
gi nui cy, hot tnh ca amylase cn rt thp. C th sau 36 gi nui cy, A. niger
bt u to bo t, nn hm lng protein gim v hot tnh amylase gim nhanh.
34
Khi nui vi sinh vt thu enzyme amylase, c hai qu trnh lin quan mt thit
vi nhau, l qu trnh tng hp sinh khi vi sinh vt v qu trnh tch ly enzyme
trong mi trng. V trong thc t, hai qu trnh ny khng h khp vi nhau v mt
thi gian.
Theo nhiu kin, s to thnh amylase cc i thng xy ra sau khi qun th
t bo t nh sinh trng. Trong qu trnh sinh trng, vi sinh vt ch tng hp nn
mt lng amylase ngoi bo rt t, v ch sau khi kt thc pha sinh trng mi xy ra
s tng hp enzyme amylase cc ln. Ngi ta cho rng s tng hp enzyme amylase
bt u khi m vic to protein ca t bo kt thc hoc gn kt thc v cnh tranh
vi qu trnh ny.
Thi gian nui to lng amylase cc ln thng c xc nh bng thc
nghim. Ty thuc tnh cht sinh sinh l ca tng vi sinh vt v s ngng tng hp
enzyme m c th ngng sinh trng ca nm mc vo bt k lc no thy cn thit.
S to bo t l hin tng khng mong mun v thng lm gim hot lc ca
enzyme. i vi a s nm mc Aspergillus s to enzyme cc i thng kt thc
khi nm mc bt u sinh nh bo t.
4.1.5. nh hng ca nhit
Trong th nghim ny, chn t l cm go: tru: b u nnh theo t l 2:1:1 v
m 55%. Bng cch thm nc ct v mi trng c nng dinh dng X1 v
iu chnh pH ca chng c c mi trng nui cy c m 55% v c pH 5.
Cc bc tip theo thc hin nh mc 3.3.6. Cy dch huyn ph bo t A. niger vi
mt 0,8*108 bo t/g vo cc mi trng chun b trn.
em nui cy trong cc t sy vi cc nhit nh sau: 30 0C, 350C, 400C,
450C, 500C v 550C. Thu dch chit enzyme t canh trng sau 28 gi nui cy, em
xc nh hot tnh v hm lng enzyme amyase thu c. Kt qu phn tch phng
sai ANOVA cho thy s khc nhau gia cc nghim thc l c ngha.
35
th 4.4 Biu din hot tnh amylase v hm lng protein t canh trng nui cy
A. niger vi cc nhit nui cy khc nhau.
mi trng nui cy trong nhit 350C cho kt qu hm lng protein v
hot tnh amylase cao nht (631,443 UI/g CT v 55,386 mg/g), v nhit nui cy
300C canh trng cho hm lng protein v hot tnh amylase cng kh cao (580,875
UI/g CT v 52,449 mg/g). Vy nhit ti u cho s tng hp amylase A. niger l
t 30 350C. Khong nhit ny thch ng vi s tng trng v pht trin ca nm
mc do , nm mc tng hp enzyme tt nht v t gi tr cc i. Cn nhit t
40 550C do nm mc khng thch nghi c nn kh nng pht trin km dn n
kh nng tng hp enzyme thp. C th do nhit trn 400C l qu cao v chng nh
hng n qu trnh sinh trng v pht trin ca nm mc A. niger.
Ton b chu k sinh trng ca nm mc trn cm c th chia thnh 3 thi k.
u tin l thi k trng mm ca nh bo t. Trong thi k ny phi duy tr nhit
trong khong 30 - 320C. Tip n l thi k sinh trng nhanh ca h si. giai on
ny, nm mc h hp rt mnh v sinh ra mt lng nhit rt ln. Kt qu l trong lp
si nm ang mc nhit tng ln n 37 - 400C. V vy trong lc ny cn ch
gim nhit phng gip si nm mc u v p. Cui cng l thi k to amylase
mnh m. Trong thi k ny cc qu trnh trao i cht dn dn yu i, s ta nhit
gim mnh, cc enzyme amylase c tng hp mnh m. Trong giai on ny, nn
h nhit thp hn nhit ca giai on u t 3 - 40C. Nhn chung, nhit thch
hp nm si sinh trng v tng hp amylase trn mi trng rn nm trong
khong t 28 350C.
36
th 4.5 Biu din hot tnh amylase v hm lng protein t canh trng nui cy
A. niger vi cc m mi trng ban u khc nhau.
m mi trng thch hp cho s sinh trng v pht trin ca A. niger l
55%. mi trng c m ny hm lng protein v hot lc enzyme amylase thu
c l cao nht (583,331 UI/g CT v 55,375 mg/g). Trong khi mi trng c m
t 40% n 50% c hot lc amylase v hm lng protein thp hn nhiu. iu ny
c th do tng m thch hp c tc dng lm c cht phng ln, tng xp to
iu kin cho nm d dng tip xc vi c cht hn. Cn cc mi trng cn li,
m cng cao th c cht cng t nn kt dnh vo nhau, mi trng khng c xp
nn nh hng n kh nng tng hp enzyme ca nm mc. Hin tng ny c th
37
th 4.6 Biu din hot tnh amylase v hm lng protein t canh trng nui cy
A. niger vi cc nng dinh dng trong mi trng khc nhau.
38
Hot lc amylase cao nht thu c nng mi trng dinh dng X2.
Nhng hm lng protein thu c hi thp hn so vi mi trng c nng dinh
dng X1, nn mi trng X2, hot tnh ring ca amylase l cao nht so vi cc
mc mi trng cn li. iu ny c l l do A. niger khng chu c p sut thm
thu cao t mi trng nui cy nn nhng mi trng c nng dinh dng qu
s cao nh hng n kh nng sinh trng v pht trin ca nm A.niger, v th m
nh hng n kh nng to enzyme ca chng.
Yu cu c bn i vi thnh phn mi trng dinh dng nui nm si to
amylase cng ging nh yu cu i vi mi trng nui vi sinh vt to enzyme l
tnh hon thin. V vy, tng cng s sinh trng ca vi sinh vt cng nh tng
kh nng to enzyme mong mun, ngoi cc thnh phn dinh dng c trong c cht,
mi trng nui cy s c b sung thm cc cht khong vi lng v a lng vn
l nhng cu t khng th thiu cho sinh trng v pht trin ca vi sinh vt.
Nng khong b sung vo mi trng phi va , v nu thp s khng
cho nhu cu ca vi sinh vt, cn nu cao, vi sinh vt s sinh trng km do khng chu
c p sut thm thu.
4.1.8. nh hng ca t l ging
Mt ging i khi cng c nh hng ng k n s sinh trng ca vi sinh
vt trong ln men bn rn. Trong th nghim ny, chn t l cm go: tru : b u
nnh theo t l 2:1:1 v m 55% kho st nh hng ca pH. Bng cch thm
nc ct v mi trng c nng dinh dng X2 v iu chnh pH ca chng c
c mi trng nui cy c m 55% v c pH 5. Cc bc tip theo thc hin nh
mc 3.3.6. Cy dch huyn ph bo t A. niger vi cc mt 0,8*108; 1,6*108;
2,4*108; 3,2*108 bo t/g vo cc mi trng chun b trn.
Thu nhn enzyme t canh trng sau 28 gi nui cy, thu dch chit enzyme t
canh trng, em xc nh hot tnh v hm lng enzyme amyase thu c. Kt qu
phn tch phng sai ANOVA cho thy s khc nhau gia cc nghim thc l c
ngha v mt thng k hc.
39
th 4.7 Biu din hot tnh amylase v hm lng protein t canh trng nui cy
A. niger vi cc t l ging khc nhau.
mi trng c t l ging l 0,8*10 8 c hot tnh v hm lng l cao nht
(619,68 IU/g CT v 51,833 mg/g). Mt bo t phi l 0,8*108 m bo cc tiu
phn c cht phi c bao ph bi h si nm nhm hn ch s tp nhim. Nu t l
ging qu cao dn n s cnh tranh v dinh dng gia cc c th nm dn n gim
s sinh trng v pht trin, nn hm lng protein v hot tnh amylase s gim theo.
Khi vi sinh vt c nui trong mi trng kn, khng c cung cp mi
trng dinh dng mi trong qu trnh , th s lng cht dinh dng s gim v
lng cht thi trong mi trng s tng ln. V vy, cng mt lng c cht ban u
nh nhau v thu hoch sau mt thi gian nui cy nh nhau, nu cc nng ging
cho vo l khc nhau s cho cc kt qu thu nhn enzyme khc nhau do kh nng sinh
trng ca qun th vi sinh vt cho vo.
Nu lng vi sinh vt cho vo thp, th thi gian thu hoch vi sinh vt cha tch
ly lng amylase trong canh trng do thi gian sinh trng t n phase cp
s qu di. Cn nu lng vi sinh vt cho vo l qu nhiu, nng dinh dng trong
qun th l khng , v vy chng s nhanh chng chuyn sang phase t vong do
thiu dinh dng v tch t c t trong mi trng.
40
(b)
(a)
41
th 4.8 Biu din hot tnh amylase v hm lng protein t canh trng nui cy
Mucor vi cc t l c cht khc nhau.
Hm lng protein thu c cao nht mi trng c t l cm: tru: b u
nnh l 1:1:2, nhng mi trng ny, hot tnh amylase thu c li l thp nht.
42
43
th 4.9 Biu din hot tnh amylase v hm lng protein t canh trng nui cy
Mucor vi cc pH mi trng khc nhau.
Hot tnh v hm lng thu c t mi trng pH5 l cao nht (23,046 mg/g
v 263,427 UI/g CT). cc mi trng c pH 4 v pH 6, hm lng protein v hot
tnh amylase cng tng i cao hn nhng mi trng cn li. Vy c l pH t 4 n
6 trong mi trng ban u l thch hp cho s sinh trng v pht trin ca Mucor.
Trong mi trng c pH 2, c gi tr hm lng v hot tnh thu c u thp, c th
do Mucor khng sinh trng c trong mi trng c tnh cht acid qua mnh.
mi trng pH 7 v 8, hot tnh amylase thu c l thp nhng hm lng protein
trong canh trng vn tng i cao do lng protein c sn trong mi trng. V t
biu trn ta cng thy c s nh hng ca pH n s ion ha ca c cht v
bn bn trong c cht.
pH c nh hng r rt ln s tng trng ca vi sinh vt. Mi loi ch pht
trin trong mt khong pH xc nh. a s nm mc pht trin trong mi trng hi
acid v chng l nhng vi sinh vt a acid thuc nhm acidophile.
44
45
th 4.10 Biu din hot tnh amylase v hm lng protein t canh trng nui cy
Mucor vi cc mc thi gian thu nhn khc nhau.
Thi gian thch hp nht thu amylase t mi trng nui cy Mucor l 32
gi sau khi cy (34,084 mg/g v 414,722 UI/g CT). Khong thi gian t sau 28 gi
cy, Mucor bt u sinh tng hp mnh m enzyme amylase. Trc , lng amylase
thu c khng nhiu v hot tnh cng khng cao. Nhng t sau 36 gi nui cy,
hot tnh amylase gim nhanh chng, song hm lng protein li gim chm hn, iu
ly c th do t sau 36 gi nui cy, Mucor chuyn sang dng bo t nn hm lng
amylase khng cn nhiu trong canh trng, dn dn hot tnh ca amylase gim. Vy
thi gian nui cy c ngha rt quan trng trong vic thu nhn amylase t mi trng
nui cy vi sinh vt.
Theo l thuyt hin i th gia vn tc sinh trng ring ca vi sinh vt v tc
sinh tng hp enzyme c mi tng quan ph thuc. Kiu ph thuc th nht l
vn tc sinh trng ca vi sinh vt hon ton ph hp chnh xc vi vn tc sinh tng
hp enzyme. Kiu ph thuc th hai l ngoi s tng hp enzyme trong pha sinh
trng, cn c s tng hp enzyme thm. S to thm ny khng lin quan t l thun
vi s sinh trng ca vi sinh vt. S to thm ny c thc hin bi cc t bo
chuyn sang qu trnh t phn v ph thuc vo bn vng ca cc ribonucleic acid
thng tin. Nh vy l kh nng khng trng khp ca cc giai on sinh trng vi
giai on sinh tng hp enzyme c xc inh bng bn ca ribonucleic acid thng
46
tin (ribonucleic acid khun). V vy m qua trnh tch ly enzyme cc i trong canh
trng ch xy ra sau qu trnh sinh sn ca nm.
Thi gian nui to lng amylase cc ln thng c xc nh bng thc
nghim. Ty thuc tnh cht sinh sinh l ca tng vi sinh vt v s ngng tng hp
enzyme m c th ngng sinh trng ca nm mc vo bt k lc no thy cn thit.
S to bo t l hin tng khng mong mun v t lm gim hot lc ca enzyme.
4.2.5. nh hng ca nhit
Trong th nghim ny, chn t l cm go: tru: b u nnh theo t l 2:1:1 v
m 55%. Thm nc ct v mi trng c nng dinh dng X1 v iu chnh
pH ca chng c c mi trng nui cy c m 55% v c pH 5. Cc bc
tip theo thc hin nh mc 3.3.6. Cy dch huyn ph bo t Mucor vi mt 8*108
bo t/g vo cc mi trng chun b trn.
em nui cy trong cc t sy vi cc nhit nh sau: 30 0C, 350C, 400C,
450C, 500C v 550C, thu dch chit enzyme t canh trng sau 32 gi, em xc nh
hot tnh v hm lng enzyme amyase thu c. Kt qu phn tch phng sai
ANOVA cho thy s khc nhau gia cc nghim thc l c ngha.
th 4.11 Biu din hot tnh amylase v hm lng protein t canh trng nui cy
Mucor vi cc nhit nui cy khc nhau.
Nhit ti u cho s sinh tng hp amylase l 300C, nhit ny hm
lng v hot tnh u cao. Chng t t l hm lng amylase trong canh trng khi
Mucor c nui 300C l tt nht. Hm lng v hot tnh amylase gim nhanh
47
chng nhng mi trng nui cy trn 400C, do nhit cao c ch kh nng sinh
trng ca Mucor. n 550C, lng protein cn rt thp, ko theo hot tnh ca
amylase trong mi trng cng rt thp.
Cng nh mi sinh vt sng, nm mc cng chu nh hng su sc ca nhit
mi trng. Chng c bit nhy cm do c cu to n bo v tnh bin nhit. V
th m c s bin thin rt mnh ca hot tnh enzyme khi c s thay i ca nhit
nui cy. Thn nhit ca c th nm phn nh trc tip nhit ca mi trng.
Mt trong nhng nhn t quan trng nht c lin quan n tc ng ca nhit
trn s tng trng, l nhit cm ng ca nhng phn ng thy phn c xc
tc bi enzyme. mc nhit thp, s tng nhit s t l thun vi s tng vn
tc tng trng. Khi hot ng bin dng s c gia tng v vi sinh vt cng s
pht trin nhanh hn. Nhng nu vt qu mt im no (im ti u), s tng
trng ca sinh vt s gim v nu tip tc tng nhit s gy cht vi sinh vt. Nhit
cao s lm cc vi sinh vt b tn thng do enzyme b bin tnh, ng thi cng nh
hng rt ln n cc h thng vn chuyn v cc protein khc. Nu cc enzyme chc
nng hot ng vi vn tc nhanh hn nhit cao vi sinh vt c th b tn thng
v s tng trng s b c ch.
S tng trng ca nm ni ring v ca vi sinh vt ni chung ph thuc c
bit vo nhit . C ba mc ca nhit sinh trng, gm nhit ti thiu, nhit ti
a v nhit ti thch. V d ng cong tng trng theo nhit c th bin i,
nhng nhit ti thch bao gi cng gn vi nhit ti a hn l nhit ti thiu.
Tuy nhin, nhit sinh trng cng cn ph thuc nhiu vo cc yu t khc ca mi
trng nui cy nh pH, cht dinh dng c sn.
4.2.6. nh hng ca m ban u
m va nh hng n sinh trng va nh hng n trao i cht ca vi
sinh vt trong qu trnh ln men bn rn. Trong th nghim ny, chn t l Cm go:
tru: b u nnh theo t l 2:1:1 kho st nh hng ca m. Thm nc ct v
mi trng c nng dinh dng X1 v pH 5 iu chnh m cc mi trng
n 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, cc bc tip theo thc hin nh
mc 3.3.6. Cy dch huyn ph bo t Mucor vi mt 0,8*108 bo t/g vo cc mi
trng chun b trn.
48
Sau 32h nui cy, thu dch chit enzyme t canh trng, em xc nh hot tnh
v hm lng enzyme amyase thu c. Kt qu phn tch phng sai ANOVA cho
thy s khc nhau gia cc nghim thc l c ngha.
th 4.12 Biu din hot tnh amylse v hm lng protein t canh trng nui cy
Mucor vi cc m mi trng ban u khc nhau.
Hot tnh amylase v hm lng protein cao nht thu c t mi trng c
m ban u l 55%. Sau m gim mnh c th do mi trng qu m nn ngn
cn s khuch tn O2 vo mi trng. Cn trc , hm lng v hot tnh li c xu
hng tng dn v khi tng m trong phm vi thch hp ko theo tc sinh trng
v tiu th O2 tng theo, nh , amylase cng c sinh ra nhiu hn.
Trong iu kin sn xut, m ban u ti thch ca mi trng ln men bn
rn cho nm si sinh trng v tng hp enzyme amylase l 55 60% v phi gi cho
vic n nh ca m trong sut qu trnh nui. m tng qu 70% s lm gim
thong kh, cn thp hn 50% s km hm s sinh trng v tng hp enzyme ca vi
sinh vt. Cn nu nui trong iu kin mi trng khng c v trng tuyt i cn
lu m mi trng sau khi cy ging khng c vt qu 60%, v cao hn na
s rt d b nhim cc vi sinh vt khc.
4.2.7. nh hng ca nng dinh dng
Do nhiu cht dinh dng c th di mc ti u thm ch khng c trong c
cht ang s dng, nn cn b sung thm cc cht dinh dng t bn ngoi khi pha
ch mi trng ln men. Trong phn ny chn t l cm go: tru: b u nnh theo t
49
l 2:1:1 v m 55%, pH 5 v cc nng dinh dng X1, X2 , X3, X4, X5, X6,
X7, X8. Cc bc tip theo thc hin nh mc 3.3.6. Cy dch huyn ph bo t
Mucor vi mt 0,8*108 bo t/g vo cc mi trng chun b trn.
Sau 32 gi nui cy, thu dch chit enzyme t canh trng, em xc nh hot
tnh v hm lng enzyme amyase thu c. Kt qu thu nhn th 4.13. Kt qu
phn tch phng sai ANOVA cho thy s khc nhau gia cc nghim thc l c
ngha v mt thng k hc.
Nng dinh dng X3 l thch hp nht cho s tng hp enzyme amylase.
nng dinh dng ny, hm lng protein v hot tnh amylase thu c u cao
nht (43,391 mg/g v 488,279 UI/g CT). Sau th hm lng v hot tnh u gim
dn, c th l do nng thm thu ca mi trng nh hng ln Mucor.
th 4.13 Biu din hot tnh amylse v hm lng protein t canh trng nui cy
Mucor vi cc nng dinh dng trong mi trng khc nhau.
Nng dinh dng X3 l thch hp nht cho s tng hp enzyme amylase.
nng dinh dng ny, hm lng protein v hot tnh amylase thu c u cao
nht (43,391 mg v 488,279 UI/g CT). Sau th hm lng v hot tnh u gim
dn, c th l do nng thm thu ca mi trng nh hng ln Mucor.
Yu cu c bn i vi thnh phn mi trng dinh dng nui nm si to
amylase cng ging nh yu cu i vi mi trng nui vi sinh vt to enzyme l
tnh hon thin. V vy, tng cng s sinh trng ca vi sinh vt cng nh tng
50
kh nng to enzyme mong mun, ngoi cc thnh phn dinh dng c trong c cht,
mi trng nui cy s c b sung thm cc cht khong vi lng v a lng vn
l nhng cu t khng th thiu cho sinh trng v pht trin ca vi sinh vt.
Vi sinh vt chu nh hng rt ln nu thay i nng thm thu ca mi
trng sng. iu ny l do n ch c phn cch vi mi trng bng mt mng
bn thm chn lc. Nu cho nng dinh dng vo mi trng qua niu hy qu t
s nh hng ln n p sut thm thu gy hi cho t bo sinh vt. Nu vi sinh vt
sng trong mi trng c nng dinh dng qu cao, nc s t c th sinh vt thot
ra noi mi trng. Khi , t bo v mng t bo s co li y l qu trnh co
nguyn sinh. V nh vy, t bo s b mt nc v mng t bo b tn thng. iu
ny lm t bo b bt hot v mt bin dng v s ngng pht trin.
Nc c sn ca vi sinh vt c th b gim, l do s tng tc vi cc phn t
cht tan hoc s hp thu ln b mt cc cht rn. Nng thm thu ca mi trng
sng c nh hng ln ln vi sinh vt v c dng din t mc nc c sn.
Cc nh vi sinh vt hc thng gi l hot ng ca nc. Cc vi sinh vt khc
nhau ng k v kh nng thch ng vi mi trng ngoi cnh c hot ngca
nc thp. Tuy nhin a s vi sinh vt ch sinh sn tt ti hot ng ca nc
khong 0,98 (xp x gi tr ca nc bin). V vy nu nng dinh dng qu cao s
l tng hot ng ca nc, nh hng n s sinh trng v tng hp enzyme.
4.2.8. nh hng ca t l ging
Mt ging i khi cng c nh hng ng k n s sinh trng ca vi sinh
vt trong ln men bn rn. Vi th nghim ny, chn t l Cm go: tru: b u nnh
theo t l 2:1:1 v m 55%.Thm nc ct v mi trng c nng dinh dng
X3, iu chnh pH ca chng c c mi trng nui cy c m 55% v c pH
5. Cc bc tip theo thc hin nh mc 3.3.6. Cy dch huyn ph bo t Mucor vi
cc mt 0,8*108; 1,6*108; 2,4*108; 3,2*108 bo t/g vo cc mi trng chun b.
Thu nhn enzyme t canh trng sau 32 gi nui cy, thu dch chit enzyme t
canh trng, em xc nh hot tnh v hm lng enzyme amyase thu c. Kt qu
phn tch phng sai ANOVA cho thy s khc nhau gia cc nghim thc l c
ngha xt v mt thng k hc.
51
th 4.14 Biu din hot tnh amylse v hm lng protein t canh trng nui cy
Mucor vi cc nng ging cho vo mi trng khc nhau.
mi trng c t l ging l 1,6*10 8 c hot tnh v hm lng l cao nht
(522,643 IU/g CT v 44,991 mg/g). Mt bo t 1,6*108 m bo cc tiu phn c
cht phi c bao ph bi h si nm nhm hn ch s tp nhim, v cng va
khng xy ra s cnh tranh v dinh dng gia cc c th nm nn khng b c ch s
sinh trng v pht trin ca chng.
S sinh trng ca vi sinh vt tri qua 4 giai on. u tin l phase tim phc.
Khi vi sinh vt c a vo mt mi trng nui cy mi s khng c s gia tng
ngay lp tc s lng hay sinh khi t bo. y l lc nhng thnh phn mi ca t
bo c hnh thnh. Sau s chuyn sang phase cp s. Trong phase ny, vi sinh
vt s pht trin v phn chia vi tc cc i. Sau qun th vi sinh vt s bc
vo phase n nh. S tng trng v dn s ca vi sinh vt s dng li. C nhiu l do
khin dn s vi sinh vt i vo phase n nh, d thy l s gii hn v cht dinh
dng v s tch t cht thi c hi. Cui cng l phase t vong, s lng t bo sng
s b gim do mi trng thiu dng cht v c t tch t. Nu lng vi sinh vt cho
vo thp, th thi gian thu hoch vi sinh vt cha tch ly lng amylase trong canh
trng do thi gian sinh trng t n phase cp s qu di. Cn nu lng vi sinh
vt cho vo l qu nhiu, nng dinh dng trong qun th khng , v vy chng
s chuyn sang phase t vong do thiu dinh dng v tch t c t trong mi trng.
52
53
(-)
pH 4
20
45%
X1
(0)
pH 5
28
55%
X2
(+)
pH 6
36
65%
X3
54
Bng 4.3 Hot lc amylase ca A. niger theo thc nghim v phng trnh hi qui
Stt th
Bin s m ha
nghim
X1
X2
X3
X4
Y
Y^
1
435.331
435.9468
2
+
395.221
395.8368
3
+
412.149
411.5328
4
+
+
384.872
384.2556
5
+
212.273
210.69645
6
+
+
210.185
208.60805
7
+
+
262.625
264.20195
8
+
+
+
259.129
260.70595
9
+
261.129
262.70595
10
+
+
356.950
358.52715
11
+
+
377.451
375.87405
12
+
+
+
352.539
350.96245
13
+
+
197.898
197.2816
14
+
+
+
188.092
187.4756
15
+
+
+
189.58
190.1964
16
+
+
+
+
157.949
158.5656
17
0
0
0
0
621.46
18
0
0
0
0
625.669
19
0
0
0
0
623.132
Phng trnh hi qui c dng:
Y^i=bo+b1X1+b2X2+b3X3+b4X4+b12X1X2+b13X1X3+b14X1X4+b23X2X3
+b24X2X4+b34X3X4+b123X1X2X3+b124X1X2X4+b134X1X3X4+b234X2X3X4
+b1234X1X2X3X4
Cc h s trong phng trnh hi qui nh sau:
16
xjiyi
i=1
bj =
16
Cc hiu ng tng tc c xc nh nh nhng hiu ng tuyn tnh
55
16
(xj xi)iyi
i=1
bjl =
16
Theo cng thc trn ta tnh c cc h s ca phng trnh hi qui nh sau:
b1 =
-2.719
b24 =
0.480
b2 =
8.701
b34 =
4.301
b3 =
-81.12
b123 =
5.291
b4 =
-30.64
b124 =
-9.624
b12 =
-8.196
b134 =
-10.88
b13 =
-3.159
b234 =
-17.7
b14 =
6.403
b1234 =
7.072
b23 =
-1.096
b0
=
290.84
nh gi v mt thng k hc ngha ca cc h s hi qui theo tiu chun Student.
-
yo =
-
623.419
n1
Phng sai ti hin:
Sth = 2.12111268
-
Phng sai cc h s:
Sth
Sbj =
Sbj
Vi mc tlt : mc p = 0,05, bc t do f = n-1 = 2)
tlt = t(0,05;2) = 4,3 (tra bng student)
56
=
=
=
=
=
=
=
=
5.127
16.408
152.975
57.776
15.456
5.957
12.074
2.067
> 4,3
> 4,3
> 4,3
> 4,3
> 4,3
> 4,3
> 4,3
<4,3
t24 =
t34 =
t123 =
t124 =
t134 =
t234 =
t1234 =
t0 =
0.906
8.110
9.979
18.149
20.526
33.378
13.336
548.459
<4,3
> 4,3
> 4,3
> 4,3
> 4,3
> 4,3
> 4,3
> 4,3
V t23 v t24 u nh hn 4,3 (t(0,05;2)) nn b23 v b24 b loi ra khi phng trnh
hi qui do khng c ngha.
Vy phng trnh hi qui nh sau:
Y^i=290.84-2.719X1+8.701X2-81.12X3-30.64X4
-8.196X1X2-3.159X1X3+7.403X1X4+4.3015X3X4
+5.291X1X2X3-9.624X1X2X4-10.88X1X3X4
-17.7X2X3X4+7.072X1X2X3X4
(4.1)
(yi - yi^)2
i=1
S2tt =
= 9.4309
N-l
vi N : s th nghim
= 16
l: s h s c ngha = 14
S2tt
Ta c:
Ftn =
= 2.09617504
2
S th
Gi tr chun Fisher mc p = 0.05; f1 = N-l; f2 = n-1 l:
F lt (0,05;2;2 )= 19,0
=> Ftn < F lt
57
Mc di
(-)
pH 4
24
45%
X1
Mc c s
(0)
pH 5
32
55%
X3
Mc trn
(+)
pH 6
40
65%
X5
X1
+
+
+
+
+
+
+
+
0
Bin s m ha
X2
X3
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
0
0
X4
+
+
+
+
+
+
+
+
0
58
X4
+
+
+
+
+
+
+
+
0
0
0
Y
384.107
295.175
193.030
172.615
218.142
161.876
179.793
100.464
255.177
177.848
149.388
145.4104
158.206
147.338
110.523
75.544
480.137
485.279
479.215
Y^
384.1358
295.147
191.8501
173.7952
218.5874
161.4314
180.5004
99.75772
253.9973
179.0282
149.4171
145.3819
158.9126
146.6313
110.9676
75.09956
Y^i=bo+b1X1+b2X2+b3X3+b4X4+b12X1X2+b13X1X3+b14X1X4+b23X2X3
+b24X2X4+b34X3X4+b123X1X2X3+b124X1X2X4+b134X1X3X4+b234X2X3X4
+b1234X1X2X3X4
Cc h s trong phng trnh hi qui nh sau:
16
xjiyi
i=1
bj =
16
59
(xj xi)iyi
i=1
bjl =
16
Theo cng thc trn ta tnh c cc h s ca phng trnh hi qui nh sau:
b1 = -23.256
b24 = 9.731
b2 = -41.944
b34 = 9.277
b3 = -38.804
b123 =
-11.815
b4 = -30.361
b124 =
0.237
b12 = 5.9184
b134 =
3.857
b13 =
0.576
b234 = -12.195
b14 = 7.362
b1234 =
-0.367
b23 = 14.539
b0
= 182.79
nh gi v mt thng k hc ngha ca cc h s hi qui theo tiu chun Student.
-
yo =
-
481.54
n1
Phng sai ti hin:
3
(yoi - yo)2
i=1
S2th =
= 10.677 (vi n = 3)
n-1
Sth = 3.2676
Phng sai cc h s:
Sth
Sbj =
Sbj
Vi mc tlt : mc p = 0,05, bc t do f = n-1 = 2
tlt = t(0,05;2) = 4,3 (tra bng student)
60
t1 = 28.468
t2 = 51.345
t3 = 47.502
t4 = 37.166
t12 = 7.245
t13 = 0.705
t14 = 9.012
t23 = 17.798
V t13, t124 v t1234 u
> 4,3
t24 = 11.912
> 4,3
t34 = 11.357
> 4,3
t123 = 14.463
> 4,3
t124 = 0.289
> 4,3
t134 = 4.721
< 4,3
t234 = 14.929
> 4,3
t1234 =
0.449
>4,3
t0 = 223.762
nh hn 4,3 (t(0,05;2)) nn b13, b124 v
>4,3
> 4,3
> 4,3
< 4,3
> 4,3
> 4,3
< 4,3
> 4,3
b1234 b loi ra khi
(4.2)
(yi - yi^)2
i=1
S2tt =
= 2.8568
N-l
vi N : s th nghim = 1, l: s h s c ngha = 13.
S2tt
Ta c:
Ftn =
= 0.2676
2
th
S
Gi tr chun Fisher mc p = 0.05; f1 = N-l; f2 = n-1 l:
F lt (0,05;3;2 )= 19,2
=> Ftn < F lt
61
62
Chng 5
KT LUN V NGH
5.1. Kt lun
Gi tr cc yu t nh hng tng i thch hp A. niger sinh tng hp
enzyme amylase l t l C:T:B (2:1:1), pH 5, thi gian nui cy 28 gi, nhit nui
cy 300C, m ban u 55%, 2 ln nng dinh dng, t l ging 0,8*108 bt/g CT.
Trong cc yu t trn cc yu t c tc ng mnh n hm lng protein v hot tnh
enzyme amylase l pH, thi gian nui cy, m v nng dinh dng. Trong
yu t m l c tc ng mnh nht.
Gi tr cc yu t nh hng tng i thch hp Mucor sinh tng hp
enzyme amylase l t l C:T:B (2:1:1), pH 5, thi gian nui cy 32 gi, nhit nui
cy 350C, m ban u 55%, 3 ln nng dinh dng, t l ging 1,6*108 bt/g CT.
Trong cc yu t trn cc yu t c tc ng mnh n hm lng protein v hot tnh
enzyme amylase l pH, thi gian nui cy, m v nng dinh dng. Trong
yu t thi gian l c tc ng mnh nht.
A. niger sinh tng hp enzyme amylase mnh hn chng Mucor, hm lng
protein v hot tnh enzyme amylas do A. niger to ra u cao hn do Mucor to ra.
Hn na, nm mc Mucor c sc sinh trng km hn A. niger v rt d b xm
nhim bi cc loi vi sinh vt khc.
5.2. ngh
- Thc hin k hoch leo dc tm ra mi trng thch hp nht cho hai chng
nm mc nghin cu to enzyme amylase mnh m nht.
- Th nghim c tnh canh trng nui cy Asp. niger v Mucor trn chut
bch.
- Th nghim lm ch phm enzyme dng trong thc n chn nui.
- Thc hin tinh sch enzyme amylase t cc canh trng nui cy trn thu
nhn enzyme tinh khit hn.
- Th nghim c nh ch phm enzyme amylase t canh trng nui cy ti u
trn c cht l Chitosan v Natrialginate.
63
Ch Minh.
2. Nguyn Ln Dng v cc tc gi. 1979. Mt s phng php nghin cu vi sinh
vt, tp 2. NXB Khoa hc v k thut H Ni.
3. Phm Th nh Hng.2004. K thut sinh ho. NXB i Hc Quc Gia
Tp.HCM.
4. Nguyn c Lng v cc tc gi.2004. Cng ngh enzyme. NXB i hc
Thng k, H Ni.
8. Trn Anh Tun. 2005. Kha st cc yu t nh hng n s sinh tng hp
enzyme lipase t Aspergillus niger trn mi trng ln men bn rn. Kha lun tt
nghip khoa sinh hc, Trng i Hc Khoa Hc T Nhin Tp.HCM
Nc ngoi
9. Amano Pharmaceutical Co.,Ltc. (APC), (2000), Assay method for amylase
64
PH LC
Hnh 3 Mucor.
K hoch leo dc
Ti u ha thc nghim bng phng php leo dc trn mt mc tiu l giai
on th hai ca qu trnh ti u ha thc nghim. Do nghin cu i t m hnh tuyn
tnh m t cc b mt mc tiu khi tin dn n min cc tr th m t tng thch ny
s khng cn ph hp nn phi tin hnh xc nh im ti u bng phng php leo
dc. Mc ch ca k hoch leo dc l tm ra ta ca cc tr ca mt mc tiu bng
thc nghim.
Ni dung: k hoch leo dc mang ni dung ca phng php lp bao gm
nhng bc c bn sau:
1. Chn im xut pht: thng mi trng ti tm th nghim s c chn
lm im xut pht.
2. Chn hng di chuyn ca gradient: t im c s xc nh mc cc yu t
v tin hnh xy dng k hoch thc nghim trc giao cp 1.
3. Di chuyn theo hng gradient (cn tm khong cch di chuyn thch hp)
4. Tm iu kin dng (trong kha lun ny, iu kin dng th nghim leo
Nghim thc
C cht
2C:1T:1B
1C:2T:1B
1C:1T:2B
1
40.417
36.075
Ln lp li
2
41.481
36.863
Trung bnh
3
42.494
37.974
41.484
36.971
40.372
41.17
42.271
41.271
Bng 1.2 Bng ANOVA cho hm lng da trn s liu bng 1.1
ANOVA Table for ham luong by ty le co chat
Source
P-Value
Between groups
Within groups
Total (Corr.)
Analysis of Variance
Sum of Squares Df
Mean Square
38.9025
5.6384
44.5409
8
2
6
19.4513
20.70
0.939734
F-Ratio
0.0020
The StatAdvisor
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
20.6987, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of ty le co chat to another at the 95.0% confidence level.
Nghim thc
C cht
2C:1T:1B
1C:2T:1B
1C:1T:2B
1
357.45
307.27
199.48
Ln lp li
2
358.44
310.26
200.47
Trung bnh
3
359.76
308.59
201.8
358.552
308.373
200.584
Bng 1.4 Bng ANOVA cho hot tnh da trn s liu bng 1.3
ANOVA Table for hoat tinh by ty le co chat
Source
Value
Between groups
0.0000
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares Df
Mean Square
39090.3
19545.1
8.07446
39098.4
8
1.34574
F-Ratio
P-
14523.67
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
14523.7, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of ty le co chat to another at the 95.0% confidence level.
Nghim thc
pH
2
3
4
5
6
7
8
1
19.934
22.709
29.705
40.861
27.531
24.465
22.224
Ln lp li
2
20.723
23.812
32.650
41.849
28.519
25.453
23.212
Trung bnh
3
21.811
24.621
30.404
43.173
29.843
26.777
24.536
20.822
23.714
30.586
41.961
28.631
25.565
23.324
Bng 2.2 Bng ANOVA cho hm lng da trn s liu bng 2.1
ANOVA Table for ham luong by pH
Source
P-Value
Between groups
0.0000
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares Df
898.19
15.8364
914.026 20
6
14
Mean Square
149.698
F-Ratio
132.34
1.13117
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
132.339, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of pH to another at the 95.0% confidence level.
Nghim thc
pH
2
3
4
5
6
7
8
1
135.36
178.22
250.59
353.23
241.02
176.77
126.22
Ln lp li
2
136.35
179.2
251.57
354.22
242.01
177.76
127.21
Trung bnh
3
137.68
180.53
252.9
355.55
243.33
179.08
128.53
136.463
179.315
251.686
354.333
242.12
177.869
127.321
Bng 2.4 Bng ANOVA cho hot tnh da trn s liu bng 2.3
ANOVA Table for hoat tinh by pH
Analysis of Variance
Source
Sum of Squares
Df
Mean Square
F-Ratio
P-Value
Between groups
113456.0
18909.4
18.8404
14
1.34574
14051.26
0.0000
Within groups
Total (Corr.)
113475.0
20
The StatAdvisor
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
14051.3, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of pH to another at the 95.0% confidence level.
Nghim thc
Ln lp li
Trung bnh
1
18.546
21.721
28.802
39.513
43.907
51.424
41.294
39.553
35.046
30.375
2
19.534
22.709
29.79
40.501
44.895
52.412
42.282
40.541
36.034
31.363
3
20.858
24.033
31.114
41.825
46.219
53.736
43.606
41.865
37.358
32.687
19.646
22.821
29.902
40.613
45.007
52.524
42.394
40.653
36.146
31.475
Bng 3.2 Bng ANOVA cho hm lng da trn s liu bng 3.1
ANOVA Table for ham luong by thoi gian nuoi cay
Source
P-Value
Between groups
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares Df
Mean Square
2809.99
26.9149 20
2836.91
29
9
312.221
1.34574
232.01
F-Ratio
0.0000
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
232.007, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of thoi gian nuoi cay to another at the 95.0% confidence
level.
Nghim thc
Thi gian nui cy (h)
8
12
16
20
24
28
32
36
40
44
1
109.83
171.14
255.84
309.32
383.74
581.16
378.88
353.51
225.14
101.73
Ln lp li
2
110.82
172.13
256.82
310.31
384.73
582.15
379.87
354.5
226.13
102.72
Trung bnh
3
112.14
173.46
258.15
311.63
386.05
583.47
381.2
355.83
227.45
104.04
110.929
172.244
256.935
310.421
384.841
582.262
379.984
354.613
226.242
102.832
Bng 3.4 Bng ANOVA cho hot tnh da trn s liu bng 3.3
ANOVA Table for hoat tinh by thoi gian nuoi cay
Source
P-Value
Between groups
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares
Df
Mean Square
579567.0
9
26.9149
579594.0
29
64396.3 47851.86
20
1.34574
F-Ratio
0.0000
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
47851.9, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of thoi gian nuoi cay to another at the 95.0% confidence level.
Nghim thc
nhit nui cy (0C)
30
35
40
45
50
55
1
51.349
54.286
43.549
36.336
23.671
15.572
Ln lp li
2
52.337
55.274
44.537
37.324
24.659
16.56
Trung bnh
3
53.661
56.598
45.861
38.648
25.983
17.884
52.449
55.386
44.649
37.436
24.771
16.67166
Bng 4.2 Bng ANOVA cho hm lng da trn s liu bng 4.1
ANOVA Table for ham luong by nhiet do nuoi cay
Source
P-Value
Between groups
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares Df
Mean Square
3550.78
16.1489
3566.93 17
5
12
710.156
527.71
1.34574
F-Ratio
0.0000
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
527.705, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of nhiet do nuoi cay to another at the 95.0% confidence level.
Nghim thc
nhit nui cy (0C)
30
35
40
45
50
55
1
579.78
630.34
408.69
267.17
176.61
89.468
Ln lp li
2
580.76
631.33
409.68
268.16
177.6
90.456
Trung bnh
3
582.09
632.66
411
269.49
178.92
91.78
Bng 4.4 Bng ANOVA cho hot tnh da trn s liu bng 4.3
580.876
631.443
409.792
268.273
177.709
90.568
Analysis of Variance
Sum of Squares
Df
717552.0
16.1489
717568.0
12
17
Mean Square
143510.0
F-Ratio
06640.15
1.34574
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
106640.0, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of nhiet do nuoi cay to another at the 95.0% confidence level.
Nghim thc
m (%)
40
45
50
55
60
65
70
75
1
36.072
42.975
49.776
54.275
51.336
47.425
41.268
36.259
Ln lp li
2
37.06
43.963
50.764
55.263
52.324
48.413
42.256
37.247
Trung bnh
3
38.384
45.287
52.088
56.587
53.648
49.737
43.58
38.571
37.172
44.0752
50.876
55.375
52.436
48.525
42.368
37.3589
Bng 5.2 Bng ANOVA cho hm lng da trn s liu bng 5.1
ANOVA Table for ham luong by do am
Source
P-Value
Between groups
Within groups
Total (Corr.)
Analysis of Variance
Sum of Squares Df
Mean Square
986.874
21.5319
1008.41 23
7
16
140.982
104.76
1.34574
F-Ratio
0.0000
The StatAdvisor
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
104.761, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of do am to another at the 95.0% confidence level.
Nghim thc
m (%)
40
45
50
55
60
65
70
75
1
198.92
331.62
408.26
582.23
266.66
168.71
155.25
142.26
Ln lp li
2
199.91
332.6
409.24
583.22
267.65
169.7
156.24
143.25
Trung bnh
3
201.23
333.93
410.57
584.54
268.97
171.02
157.56
144.57
200.021
332.716
409.356
583.331
267.758
169.812
156.351
143.358
Bng 5.4 Bng ANOVA cho hot tnh da trn s liu bng 5.3
Source
Value
Between groups
0.0000
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares
Df
Mean Square
492316.0
21.5319
492337.0
16
23
70330.8
F-Ratio
P-
52261.66
1.34574
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
52261.7, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of do am to another at the 95.0% confidence level.
Nghim thc
Nng dinh dng
X1
X2
X3
X4
X5
X6
X7
X8
Ln lp li
2
54.515
53.301
49.836
48.953
45.067
40.855
40.569
37.992
1
53.527
52.313
48.848
47.965
44.079
39.867
39.581
37.004
Trung bnh
3
55.839
54.625
51.16
50.277
46.391
42.179
41.893
39.316
54.627
53.413
49.948
49.065
45.179
40.967
40.681
38.104
Bng 6.2 Bng ANOVA cho hm lng da trn s liu bng 6.1
Analysis of Variance for ham luong - Type III Sums of Squares
Source
Sum of Squares
Df
Mean Square
F-Ratio
P-Value
MAIN EFFECTS
A:nong do dinh duong
807.055
115.294
RESIDUAL
21.5319
16
TOTAL (CORRECTED)
828.587
23
85.67
0.0000
1.34574
Since Type III sums of squares (the default) have been chosen, the
contribution of each factor is measured having removed the effects of all other factors. The Pvalues test the statistical significance of each of the factors. Since one P-value is less than
0.05, this factor has a statistically significant effect on ham luong at the 95.0% confidence
level.
Nghim thc
Nng dinh dng
X1
X2
X3
X4
X5
X6
X7
X8
Ln lp li
2
576.71
624.86
547.18
461.53
416.78
378.52
373.77
338.01
1
575.72
623.87
546.19
460.54
415.79
377.53
372.79
337.02
Trung bnh
3
578.03
626.18
548.51
462.85
418.1
379.84
375.1
339.34
576.821
624.969
547.294
461.642
416.887
378.631
373.885
338.123
Bng 6.4 Bng ANOVA cho hot tnh da trn s liu bng 6.3
ANOVA Table for hoat tinh by nong do dinh duong
Analysis of Variance
Source
Sum of Squares
Df
Mean Square
F-Ratio
P-Value
Between groups
237154.0
33879.2
Within groups
21.5319
16
1.34574
Total (Corr.)
237176.0
23
25175.05
0.0000
The StatAdvisor
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
25175.0, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of nong do dinh duong to another at the 95.0% confidence
level.
Nghim thc
T l ging (*108bt/g)
0.8
1.6
2.4
3.2
1
50.733
47.592
43.317
42.514
Ln lp li
2
51.721
48.48
44.805
43.502
Trung bnh
3
53.045
49.704
46.629
44.826
51.833
48.592
44.917
43.614
Bng 7.2 Bng ANOVA cho hm lng da trn s liu bng 7.1
ANOVA Table for ham luong by ty le giong
Source
Value
Between groups
0.0001
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares
Df Mean Square
124.403
10.766
135.169 11
3
8
41.4678
F-Ratio
P-
30.81
1.34574
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
30.814, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of ty le giong to another at the 95.0% confidence level
Nghim thc
T l ging (*108bt/g)
0.8
1.6
2.4
3.2
1
618.58
546.96
434.47
404.75
Ln lp li
2
619.57
547.95
435.45
405.74
Bng 7.4 Bng ANOVA cho hot tnh da trn s liu bng 7.3
ANOVA Table for hoat tinh by ty le giong
Analysis of Variance
Trung bnh
3
620.89
549.27
436.78
407.07
619.678
548.061
435.566
405.853
Source
P-Value
Between groups
0.0000
Within groups
Total (Corr.)
The StatAdvisor
Sum of Squares
Df
88881.3
Mean Square
29627.1
1.34574
10.766
88892.1 11
F-Ratio
22015.42
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
22015.4, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of ty le giong to another at the 95.0% confidence level.
Nghim thc
T l c cht
2C:1T:1B
1C:2T:1B
1C:1T:2B
Ln lp li
2
21.181
18.564
24.995
1
19.351
16.734
23.165
Trunng bnh
3
21.421
18.804
25.235
20.651
18.034
24.465
Bng 8.2 Bng ANOVA cho hm lng da trn s liu bng 8.1
ANOVA Table for ham luong by ty le co chat
Source
Value
Between groups
0.0013
Within groups
Total (Corr.)
The StatAdviso
Analysis of Variance
Sum of Squares
Df
Mean Square
62.753
2
7.6914
70.4444
F-Ratio
31.3765
6
P-
24.48
1.2819
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
24.4766, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of ty le co chat to another at the 95.0% confidence level.
Nghim thc
Ln lp li
Trunng bnh
T l c cht
2C:1T:1B
249.699
251.529
251.769
250.999
1C:2T:1B
224.825
226.655
226.895
226.125
1C:1T:2B
170.2
172.03
172.27
171.5
Bng 8.4 Bng ANOVA cho hot tnh da trn s liu bng 8.3
ANOVA Table for hoat tinh by ty le co chat
Analysis of Variance
Sum of Squares Df Mean Square
Source
Value
Between groups
Within groups
Total (Corr.)
The StatAdvisor
9922.7
2
4961.35 3870.31
7.6914
6
1.2819
9930.39
8
F-Ratio
P-
0.0000
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
3870.31, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of ty le co chat to another at the 95.0% confidence level. To
determine
Nghim thc
pH
2
3
4
5
6
7
8
1
6.883
11.83
17.938
21.746
17.271
13.742
13.023
Ln lp li
2
8.713
13.66
19.768
23.576
19.101
15.572
14.853
Trunng bnh
3
8.953
13.9
20.008
23.816
19.341
15.812
15.093
8.183
13.13
19.238
23.046
18.571
15.042
14.323
Bng 9.2 Bng ANOVA cho hm lng da trn s liu bng 9.1
ANOVA Table for ham luong by pH
Source
Between groups
Sum of Squares
419.346
Analysis of Variance
Df
Mean Square F-Ratio
6
69.8911
54.52
P-Value
0.0000
Within groups
Total (Corr.)
The StatAdvisor
17.9466
437.293
14
20
1.2819
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
54.5215, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of pH to another at the 95.0% confidence level.
Nghim thc
pH
2
3
4
5
6
7
8
Ln lp li
2
90.685
155.244
205.197
263.957
170.325
111.039
86.139
1
88.855
153.414
203.367
262.127
168.495
109.209
84.309
Trunng bnh
3
90.925
155.484
205.437
264.197
170.565
111.279
86.379
90.155
154.714
204.667
263.427
169.795
110.509
85.609
Bng 9.4 Bng ANOVA cho hot tnh da trn s liu bng 9.3
ANOVA Table for hoat tinh by pH
Analysis of Varianc
Source
Sum of Squares
Df
Mean Square
F-Ratio
Value
Between groups
76308.8
Within groups
17.9466
14
Total (Corr.)
76326.8
20
12718.1
1.2819
9921.32
0.0000
P-
The StatAdvisor
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
9921.32, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of pH to another at the 95.0% confidence level. To determine
which means
Ln lp li
Trunng bnh
1
12.857
15.159
17.063
20.619
2
14.686
16.983
19.992
22.449
3
14.925
17.226
20.365
22.689
14.155
16.456
19.163
21.919
24
28
32
36
40
44
22.472
25.295
32.784
22.009
19.013
16.365
24.302
27.125
34.614
23.839
24.743
18.195
24.542
27.365
34.854
24.079
20.183
18.435
23.772
26.595
34.084
23.309
21.313
17.665
Bng 10.2 Bng ANOVA cho hm lng da trn s liu bng 10.1
ANOVA Table for ham luong by thoi gian nuoi cay
Source
Value
Between groups
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares Df
Mean Square
874.031
25.638
899.669
9
20
29
97.1146
1.2819
F-Ratio
75.76
P0.0000
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
75.7583, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of thoi gian nuoi cay to another at the 95.0% confidence
level.
Trunng
Nghim thc
Thi gian nui cy (h)
8
12
16
20
24
28
32
36
40
44
Ln lp li
2
69.038
108.485
155.163
203.190
232.060
270.018
415.252
250.394
139.631
47.705
1
67.312
106.655
153.333
201.764
230.111
268.188
413.422
249.564
137.901
45.875
bnh
3
69.48
108.725
155.403
203.638
232.242
270.258
415.492
255.634
140.071
47.945
68.61
107.955
154.633
202.864
231.471
269.488
414.722
251.864
139.201
47.175
Bng 10.4 Bng ANOVA cho hot tnh da trn s liu bng 10.3
ANOVA Table for hoat tinh by thoi gian nuoi cay
Source
P-Value
Between groups
0.0000
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares
Df
Mean Square
324641.0
25.638
324666.0 29
36071.2
20
1.2819
F-Ratio
28138.86
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
28138.9, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of thoi gian nuoi cay to another at the 95.0% confidence level.
Nghim thc
Ln lp li
Trunng
bnh
Nhit nui cy (0C)
40
45
50
55
60
65
70
75
1
23.860
27.958
33.121
38.656
33.986
30.633
23.667
22.187
2
25.691
29.793
34.951
40.486
35.816
32.463
25.407
24.017
3
25.932
29.926
35.191
40.726
36.056
32.703
25.827
24.257
25.161
29.159
34.421
39.956
35.286
31.933
24.967
23.487
Bng 11.2 Bng ANOVA cho hm lng da trn s liu bng 11.1
ANOVA Table for ham luong by nhiet do
Analysis of Variance
Source
Sum of Squares
Df
Mean Square
F-Ratio
P-
Value
Between groups
1176.66
Within groups
15.3828
12
Total (Corr.)
1192.04
17
235.332
183.58
0.0000
1.2819
The StatAdvisor
The ANOVA table decomposes the variance of ham luong into two components: a
between-group component and a within-group component. The F-ratio, which in this case
equals 183.581, is a ratio of the between-group estimate to the within-group estimate. Since
the P-value of the F-test is less than 0.05, there is a statistically significant difference between
the mean ham luong from one level of
nhiet do to another at the 95.0% confidence level.
Trunng
Nghim thc
Nhit nui cy (0C)
40
45
50
55
60
65
70
75
1
23.861
27.859
33.121
38.656
Ln lp li
2
25.691
29.089
34.951
39.486
bnh
3
25.931
30.829
35.191
41.726
25.161
29.159
34.421
39.956
33.985
30.633
23.667
22.186
35.817
32.460
25.497
24.015
36.056
32.706
25.737
24.258
35.286
31.933
24.967
23.487
Bng 11.4 Bng ANOVA cho hot tnh da trn s liu bng 11.3
Analysis of Variance
Source
Between groups
Within groups
Total (Corr.)
Sum of Squares
Df
P-Value
105956.0
0.0000
15.3828
12
1.2819
529797.0
17
529782.0
82655.72
The StatAdvisor
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
82655.7, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of nhiet do to another at the 95.0% confidence level.
Nghim thc
m (%)
40
45
50
55
60
65
70
75
1
23.863
27.861
33.121
37.656
33.987
30.036
23.667
22.187
Ln lp li
2
25.691
29.690
34.951
40.986
35.815
32.460
25.497
24.017
Trunng bnh
3
25.928
29.932
35.191
41.226
36.056
33.303
25.737
24.257
25.161
29.159
34.421
39.956
35.286
31.933
24.967
23.487
Bng 12.2 Bng ANOVA cho hm lng da trn s liu bng 12.1
Analysis of Variance
Sum of Squares Df
Mean Square
Source
F-Ratio
PValue
Between groups
423.243
7
60.4633
47.17
0.0000
Within groups
20.5104 16
1.2819
Total (Corr.)
443.754 23
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
47.167, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of do am to another at the 95.0% confidence level.
Nghim thc
m (%)
40
45
50
55
60
65
70
75
1
164.261
264.405
284.545
409.743
165.133
142.998
116.833
115.695
Ln lp li
2
164.091
266.235
286.775
411.573
166.963
144.828
117.663
117.525
Trunng bnh
3
165.331
266.475
286.815
411.813
167.203
145.068
119.903
117.765
Bng 12.4 Bng ANOVA cho hot tnh da trn s liu bng 12.3
164.561
265.705
286.045
411.043
166.433
144.298
118.133
116.995
Sum of Squares
Df
Mean Square
F-Ratio
P-Value
Between groups
69808.4
9972.63
Within groups
20.5104
16
1.2819
Total (Corr.)
69828.9
7779.57
0.0000
23
The StatAdvisor
The ANOVA table decomposes the variance of hoat tinh into two components: a
between-group component and a within-group component. The F-ratio, which in this case
equals 7779.57, is a ratio of the between-group estimate to the within-group estimate. Since
the P-value of the F-test is less than 0.05, there is a statistically significant difference between
the mean hoat tinh from one level of do am to another at the 95.0% confidence level.
Trunng
Nghim thc
Nng dinh dng
X1
X2
X3
X4
X5
X6
X7
X8
1
38.207
38.781
42.091
37.3871
35.591
32.634
30.117
29.316
Ln lp li
2
40.037
40.611
43.921
39.2171
37.421
34.464
31.947
30.146
bnh
3
40.577
40.851
44.161
39.4571
37.661
34.704
32.187
32.386
39.607
40.081
43.391
38.6871
36.891
33.934
31.417
30.616
Bng 13.2 Bng ANOVA cho hm lng da trn s liu bng 13.1
ANOVA Table for ham luong by nong do dinh duong
Analysis of Variance
Source
Sum of Squares
Df
Mean Square
F-Ratio
P-Value
Between groups
423.243
60.4633
Within groups
20.5104
16
1.2819
Total (Corr.)
443.754
23
47.17
0.0000
The StatAdvisor
The ANOVA table decomposes the variance of ham luong into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
47.167, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean ham luong from one level of nong do dinh duong to another at the 95.0% confidence
level.
Nghim thc
Nng dinh dng
X1
X2
X3
X4
X5
X6
X7
X8
1
402.621
417.348
486.979
417.529
410.567
389.136
334.487
295.962
Ln lp li
2
404.451
418.078
488.809
419.359
412.397
389.466
336.317
297.792
Trunng
bnh
3
404.691
419.918
489.049
419.599
412.637
391.206
336.557
298.032
403.921
418.548
488.279
418.829
411.867
389.936
335.787
297.262
Bng 13.4 Bng ANOVA cho hot tnh da trn s liu bng 13.3
ANOVA Table for hoat tinh by nong do dinh duong
Source
P-Value
Between groups
Within groups
Total (Corr.)
Analysis of Variance
Sum of Squares
Df
Mean Square
69808.4
20.5104 16
69828.9 23
7
9972.63
1.2819
7779.57
F-Ratio
0.0000
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
7779.57, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of nong do dinh duong to another at the 95.0% confidence
level.
Nghim thc
T l ging (*108bt/gCT)
0.8
1.6
2.4
3.2
1
38.398
43.691
37.474
33.423
Trunng
bnh
Ln lp li
2
40.228
45.021
38.304
35.253
3
40.468
46.261
40.544
35.493
39.698
44.991
38.774
34.723
Bng 14.2 Bng ANOVA cho hm lng da trn s liu bng 14.1
Source
P-Value
Between groups
Within groups
Total (Corr.)
Analysis of Variance
Sum of Squares
Df
Mean Square
160.585
10.2552
8
170.841 11
3
53.5284
1.2819
41.76
F-Ratio
0.0000
The ANOVA table decomposes the variance of ham luong into two components: a between-group
component and a within-group component. The F-ratio, which in this case equals 41.7571, is a ratio of
the between-group estimate to the within-group estimate. Since the P-value of the F-test is less than
0.05, there is a statistically significant difference between the mean ham luong from one level of ty le
giong to another at the 95.0% confidence level.
Nghim thc
T l ging (*108bt/gCT)
0.8
1.6
2.4
3.2
1
485.805
520.343
423.038
380.165
Ln lp li
2
487.635
523.173
424.868
381.995
Trunng
bnh
3
487.875
524.413
425.108
382.235
487.105
522.643
424.338
381.465
Bng 14.4 Bng ANOVA cho hot tnh da trn s liu bng 14.3
Source
Value
Between groups
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares Df
Mean Square
35846.7
10.2552
35857.0 11
3
8
11948.9
1.2819
F-Ratio
9321.25
P0.0000
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup The F-ratio, which in this case equals 9321.25, is a ratio of the between-group estimate
to the within-group estimate. Since the P-value of the F-test is less than 0.05, there is a
statistically significant difference between the mean hoat tinh from one level of ty le giong to
another at the 95.0% confidence level.
STT th nghim
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
1
388.9
298.97
217.32
175.51
222.94
167.27
183.59
104.36
268.97
166.45
155.18
151.12
160
151.13
107.32
87.337
483.14
Ln lp li
2
389.8
299.87
218.23
176.41
223.84
168.17
184.49
105.26
269.87
167.36
156.08
152.02
160.9
152.03
108.22
88.24
485.22
3
389.98
300.05
218.4
176.59
224.01
168.35
184.66
105.43
270.05
167.53
156.26
152.19
161.08
152.21
108.39
88.414
487.28
Trung bnh
389.56
299.63
217.98
176.17
223.6
167.93
184.25
105.02
269.63
167.11
155.84
151.78
160.66
151.79
107.98
87.997
485.21
Bng 15.2 Bng ANOVA cho hm lng da trn s liu bng 15.1
ANOVA Table for hoat tinh by STT
Analysis of Variance
Sum of Squares
Df
Mean Square
Source
P-Value
etween groups
Within groups
Total (Corr.)
The StatAdvisor
709835.0 16
44364.7 123691.54
12.1948 34
0.358672
709847.0 50
F-Ratio
0.0000
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
123692.0, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of STT to another at the 95.0% confidence level.
STT th
nghim
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
Y1
434.67
384.56
411.49
384.21
211.61
209.52
261.97
258.47
260.47
356.29
376.79
351.88
197.24
187.43
188.92
157.29
623.46
Ln lp li
Y2
435.57
385.46
412.39
385.11
212.52
210.43
262.87
259.37
261.37
357.19
377.69
352.78
198.14
188.33
189.82
158.19
624.67
Y3
435.75
385.64
412.57
385.29
212.69
210.6
263.04
259.55
261.55
357.37
377.87
352.96
198.31
188.51
190
158.37
625.13
Trung bnh
435.33
385.22
412.15
384.87
212.27
210.18
262.63
259.13
261.13
356.95
377.45
352.54
197.9
188.09
189.58
157.95
624.42
Bng 16.2 Bng ANOVA cho hot tnh da trn s liu bng 16.1
ANOVA Table for hoat tinh by STT
Source
Value
Between groups
0.0000
Within groups
Total (Corr.)
The StatAdvisor
Analysis of Variance
Sum of Squares
Df
Mean Square
520282.0
19.2747
520301.0
34
50
16
32517.6
F-Ratio
P-
57360.01
0.566904
The ANOVA table decomposes the variance of hoat tinh into two components: a betweengroup component and a within-group component. The F-ratio, which in this case equals
57360.0, is a ratio of the between-group estimate to the within-group estimate. Since the Pvalue of the F-test is less than 0.05, there is a statistically significant difference between the
mean hoat tinh from one level of STT to another at the 95.0% confidence level.