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Lactase Và NG D NG
Lactase Và NG D NG
Lactase Và NG D NG
NI DUNG
1. Lactose v enzyme Lactase
2. Gii thiu v Aspergillus Oryzae 3. Nui cy A.oryzae thu nhn Lactase 4. ng dng ca Lactase
LACTOSE
C IM
Cng thc phn t C12H22O11
Phn t khi
Hnh dng Khi lng ring Nhit nng chy
342.30 g/mol
Tinh th mu trng 1.525 g/cm3 202.8 oC 668.9 oC 21.6 g/100 ml
3
Lactose Nhit si
tan/ nc
ENZYME LACTASE
b galactosidase (cn gi l Lactase) l enzyme thy phn ng lactose to thnh ng glucose v ng galactose.
ENZYME LACTASE
NI DUNG
NI DUNG
1. Lactose v enzyme Lactase
2. Gii thiu v Aspergillus Oryzae 3. Nui cy A.oryzae thu nhn Lactase 4. ng dng ca Lactase
A. oryzae tit ra mi trng cc enzyme thy phn nh pectinase, xylanase, cellulase, b galactosidase,khi sng trn mi trng c ngun c cht tng ng: pectin, xylan, cellulose, lactose,...
A. oryzae c ng dng rng ri trong: Trong cng ngh sn xut thc phm: nc tng, miso, ru sake. Trong cng ngh sn xut enzyme: cellulase, amylase, protease, hemicellulase v b galactosidase.
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Nm 2001, b gene di truyn ca A. oryzae c phn tch v gii m. H gene gm 8 nhim sc th vi 12 ngn gene v 37 triu cp base
(Galagan et al., 2005, Machida et al., 2005)
NI DUNG
NI DUNG
1. Lactose v enzyme Lactase
2. Gii thiu v Aspergillus Oryzae 3. Nui cy A.oryzae thu nhn Lactase 4. ng dng ca Lactase
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NGUYN VT LIU
MI TRNG GI GING PDA Glucose 20g Agar 20g Dch chit khoai ty 1 lit pH 7 MI TRNG HOT HA Lactose 20g Peptone 3g KH2PO4 1g NaNO3 0,5g KCl 0,5g H2O 1 lit
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TUYN CHN S B
Hot b - galactosidase Chng Vi sinh vt Ngoi bo (mU/ml) Ni bo (mU/g)
A.oryzae 1
8,96
70,3
Chng A. oryzae 3 c la chn nghin cu A.oryzae 2 9,84 81,87 thu nhn enzyme ni bo
A.oryzae 3 21,24 307,5
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MT1
10 10 10 5 3 1 0.5
MT2
40 0 1 5 3 1 0.5
MT3
5 0 5 3 5 0 0.4
MT4
Mi trng Lactose c bn
CaCO3(g)
H2O(ml) pH
0
1000 7
0.2
1000 6
0
1000 7
25
pH
6,76
mU/g
0
pH
7,62
mU/g
111,61
pH
8,22
mU/g
367,01
pH
8,20
mU/g
529,47
pH
8,45
mU/g
128,58
MT1
MT2
6,15
Chn6,01 120,88 6,17 194,44 5,72 nui cy MT4 lm mi trng 285,34 5,98 0
0 6,88 45,32 7,05 165,00 6,76 180,01 6,88
169,02
MT3
6,79
277,4
MT4
6,20
43,57
6,53
264,13
6,79
398,61
6,78
507,00
6,78
738,80
26
24
36
48
60
72
Lactose
Saccarose
45.16
0
257.00
0
406.51
2.01
592.96
35.47
729.34
33.48
Maltose
Glucose Galactose
0
0 16.87
34.26
26.64 136.69
37.45
31.37 254.69
22.59
50.55 36.84
23.19
16.23 10.52
Hot b galactosidase khi nui cy A.oryzae 3 trn mi trng c ngun 27 Carbon khc nhau
Nng Lactose
Thi gian (gi) N. Lactose (g/l)
15 20 25 30
35
77.00
226.31
417.88
518.16
787.75
36
48
60
72
2g peptone + 1g (NH4)2SO4
111.81
225.48
311.73
643.00
931.41
2g peptone + 1g NH4NO3
3g cao nm men 3g peptone
113.33
56.28 65.82 34.02
346.62
175.84 177.48 61.53
529.63
220.81 231.87 82.50
727.11
351.33 561.87 108.75
1017.60
537.00 815.00 165.82 29
Hot galactosidase khi nui cy A.oryzae 3 trn mi trng c ngun nit khc nhau
3g NH4NO3
67.02
136.73
333.01
451.74
674.61
182.02
119.22
481.26
328.70
627.96
548.10
994.83
781.26
1311.80
1008.71
3g peptone + 1.5g 103.33 304.66 490.68 692.08 969.25 NH4NO3 Chn ngun v nng Nit: 1g peptone + 0,5g NH4NO3 4g peptone + 2g NH4NO3
90.62
176.24
390.42
683.93
828.38
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MI TRNG 4
Thnh phn
Lactose Peptone NH4NO3 KH2PO4 MgSO4 KOH H2O 30g 1g 0,5g 1g 0,5g 0,5g 1000ml
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nh hng pH u
Thi gian (gi) pH u
8.0
140.57
267.58
475.44
653.01
723.73
NG THI QU TRNH LN MEN MT4, 200 v/ph 300C pH 7 Sau mi 12h thu 100ml dch nm mc
o pH
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KT LUN
Tuyn chn 3 chng c kh nng sinh tng hp b- galactosidase. cao nht l A. oryzae 3 Xc nh mi trng ph hp MT4 iu kin ti u [C] Lactose 30g/l [N] Peptone 1g/l + NH4NO3 0,5 g/l pH u 7 Thi gian ln men 72 h
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Solid-state fermentation(SSF)
D thc hin Quy trnh Cng ngh khng phc tp Lng enzyme to thnh ln Ch phm enzyme th d sy kh v bo qun D x l nhim Din tch b mt ln, kh c gii ha
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Ging VSV
A. oryzae phn lp t t nhim cht thi ca nh my sa Gi ging [PDA] Nui cy bo qun 40C
Chun b mi trng
K thut nui cy
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Ln men b mt
Trn 60g cm la m v v tru (t l 1:1) vo bnh thu tinh borosil 3000 ml Lm m vi nc ct v trng cha [ Lactose 12% w/w - NaNO3 1% w/w ] [ pH u 5.0 ] Tit trng mi trng cy chuyn 10ml huyn ph cha bo t nm si [ 30oC trong 6 ngy ]
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nh lng hot bng oNPG (Sigma Chemical USA) 1 n v hot = lng enzyme gii phng
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NH HNG CA M
pH
42
Ti u ha iu kin
Ngun C Glucose cho hoat tnh enzyme mnh nht 386.6 mmol oNP/ml/pht
NaNO3> peptone> (NH4)2SO4 > urea >NH4NO3>Corn steep liquor m 90%
Ngun N
pH v nhit
Tho lun
Hot tnh enzyme lactase t cc vi sinh vt khc So snh phng php ln men & hiu qu enzyme c nh v t do iu kin nui cy nh hng ln ti hot tnh enzyme ca vi sinh vt iu kin ti u nui cy VSV thu enzyme c th khng ging vi iu kin hot ng ti u ca enzyme kt lun
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Ly tm
Tch VSV Tp ko tan
Tinh sch
Enzyme tinh khit
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Trch ly
Lc
Ly phn dch
Kt ta Sy
Sy: d bo qun
Tinh ch kt ta
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SN XUT ENZYME
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Enzymes t nm si
iu kin ti u: pH [ 3 5 ] - nhit [ 55 60C ] Khng tinh khit. Do , ng dng ca lactase t nm si gii hn cc sn phm c acid cao cng nh cc dc phm. Enzyme b- galactosidase thng mi A. oryzae: Enzeco Fungul Lactase
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NI DUNG
NI DUNG
1. Lactose v enzyme Lactase
2. Gii thiu v Aspergillus Oryzae 3. Nui cy A.oryzae thu nhn Lactase 4. ng dng ca Lactase
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Enzyme thng mi ch yu chit xut t Nm men: Kluyveromyces lactis, K. fragilis, K. marxianus, Candida kefyr Nm si: Aspergillus niger , A. oryzae Lactase l mt trong s t cc enzyme c sn xut quy m ln vi c 2 dng c nh v t do.
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NG DNG LACTASE
1981 Kligerman: Cho enzyme Lactase vo ly sa,
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Tc dng ca GOS
Tng sc khng vi vi khun gy bnh ng rut Kch thch min dch ng tiu ha Tng hp thu Canxi Qu trnh ln men sinh ra kh hi v nc phn mm v xp phng chng to bn
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Whey - casein
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Whey - casein
Ti sao sa cn c m Whey v Casein??
Whey
Casein
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Nguyn nhn
Bm sinh
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60
Xt nghim chn on
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Gii php
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Sa u nnh c lng protein cao gn bng sa b, nhng t canxi hn sa b. Sa u nnh khng c lactose.
Sa go khng c gluten Lng asen trong sa go khuyn co khng nn cho tr di 4,5 tui ung sa 63 go.
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Kt lun
b galactosidase m ra nhiu hng nghin cu mi. Nhng nghin cu ny c tim nng ng dng vo nhiu lnh vc khc nhau: cng ngh thc phm y hc Cng ngh c nh enzyme ng mt vai tr ln trong ch bin sa v whey. Vic s dng -galactosidase c nh trong thy phn lactose l mt ch khoa hc v cng ngh ng quan tm
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CM N S LNG NGHE CA C V CC BN
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