Quy trình lên men acid citric từ Aspergillus niger

I HC AN GIANG








LP DH11SH






Aspergillus niger

 






c: 2010 - 2014

An Giang, 2013


I HC AN GIANG






LP DH11SH





Aspergillus niger

 


c: 2010 - 2014

NG DN
ThS. Nguy



1. Nguyt Lynh
2. Bung San Ny Thanh Hng Anh
3. 
4. Nguyn Vit Thanh
5. 
6. Nguyn Ngc H
7. Nguyn Phm Toi Nguyn


An Giang, 2013
i
Mc lc

Ni dung Trang
Mc lc i
 iii
ng iv
t v 1
ng quan v acid citric 2
 2
i 3
t 3
ng quan v  sn xut acid citric 4
i v 4
3.2. Mt s u gluxit 4

3.2.1. Mt r 4
3.2.2. Tinh bt sn 5
 5
p cho vic sn xut acid citric 5
3.3. Vi sinh vt 5
   6
 t c 6
  6
u t n xut acid citric 6
3.5.1. Vi sinh vt 6
u 6
ng 7
ng 7
3.5.5. S  7
u t quyn xut acid citric 7
 7
ii
men 7
 7
 8
3.6.5. Th 8
 9
 9
             
 9
 9
 9
 10
 10
 10

 11
 11
4.2. Sn xut acid citric bng nm mc Aspergillus niger 11
 11
4.2.3.  12
4.2.4. Thu h acid citric 12
ng dng ca acid citric trong thc php 15
5.1. Trong thc phm 15
5.2. Trop 15
t lun ngh 16
5.1. Kt lun 16
5.2. Kin ngh 16
u tham kho 17

iii


Ni dung Trang
: Mt s sn phm cha acid citric 2
c cu to ca acid citric 3
: Mt s long acid citric c
men 4
: Aspergilus niger 11
 5: Tinh th acid citric 13
 n xut acid citric bng nm Aspergillus niger 14


iv
ng

Ni dung Trang
Bng 1: ng s dn
xut acid citric bng nm mc Aspergillus niger. 11


1
C 1
t v

c xem l t cht bo qun t  c s d b
sung v chua cho thc phi c ngt. t trong
nhng loi acid hu ng dng gn gi s
ng du cg nghip.
ng trong cid citric 
th xut hin trong i cht ca gi sinh vt, 
ch tt v ma cht ch,
ng acid a d t ra kh.
m quan tru ng dng c 
             nm mc
Aspergillus niger t u r tin nt r, tinh bt sn,
thu hi acid citric nhihn ch s dt tng
hi trong thc phi.

2

Tng quan v acid citric

2.1. 
Acid citric t acid h bin trong thc vt c
c lu (9%), trong qu  Acid citric

p thc phc gio,
 hc, dt, nhum, ngh nh, ngh in




c cam

c chanh






c lu

c da
1: Mt s sn phm cha acid citric

Acid citric    
6
H
8
O
7
. Acid citric n phm kt tinh
trong suc  th rn dy
153
0

C, t trng 1.542;  100
0
C b dung gii;  trong 150
0
C b 
acid citric khan.

3
2.2. i
Axit 2-hydroxypropan-1,2,3-tricacboxylic
ng: Axit chanh
: C
6
H
8
O
7

Kh: 192.13 g/mol
ng: Tinh th ng
Nhi y: 153
0
C
Nhi 175
o
y)



2: thc cu to ca acid citric

(Ngun:

2.3. t
 ng c - COOH, mi
         t     i
dung dm rt t hn ch s i pH c
dch acid.
t hp v ti, ph bin nh
mut bo qu v cho thc phnh
 kt hp vi t
cht bo quc.
 nhi  n ti  dng tinh th ng dng bt
hoc  da m c trong
m ca acid citric. Dc khi acid citric kt tinh trong
i dng monohydrat li kc lnh.  nhi

0
C dng monohydrat s chuyn sang dng khan.
V m              
car 
0
y t
2

c.


4
C 3
Tng quan v  sn xut acid citric

c kia acid citric ch c sn xut t u
sut thu hi thp. Hi to
citric nh nm mcitric c sn xu



3: Mt s long acid citric
c

3.1. i v
u phm b ng cho vi sinh vn.
Ging vi sinh vn xut acid citric phm bu
sau:
  o acid citric rt mnh.
  
 o ra nhng acid h
fumaric.
3.2. Mt s u gluxit
3.2.1. Mt r
Mt r t ph phm cn xun phm cua
n xu  kt tinh m
kinh t na b a mt r 
ng, ch y   i m       n
ng mt r bng khong 1/3 sng sn xut.
C khong 100 t - 4 tn r mc sn xut.

5
3.2.2. Tinh bt sn
c t trong sa nhiu trong sng,

ch yu  v ng cng 0,001 - 0,04%.
 th gii
t ch. T  
c l loi b.
S s ng HCN.
3.2.3. 
Hm hai ph       m 10 - 13% trng
ng ht, nm 83 - 85%, v chim 2 - 5%.
3.3.3. u p cho vic sn xut acid citric
t (67,9%), tit r, khoai
lang, sn cht c
ng ca vi sinh v
u tinh bt () mun sn xut ra acid citric
cn phi trn thng, t 
dng ca vi sinh vt tt mt nhiu th
trot r c ch yng vi sinh vt s trc
tip s d t.
Mt r u r tin d kit r u
c s dng nhic sn xut acid citric, tuy nhin x t
r c khi s dt r cha ch c bii
nng c ch vi sinh vn.
3.3. Vi sinh vt
Aspergillus niger t loi nm st trong nh bin nht
cAspergillus u ng d sinh hc, nhi
t 35 - 37
0
C, pH t 1,4 - c gm
mt s loqu u ph
t chm ph bin ca thc phm.
Aspergillus niger t hit h

 y g khp mt.
y trong cht thu thc v
ng o ra mt
ng sng tt cho s n ca nm mc. Chng h
c h m hoc
khu vc khu vc   mositure.
6
3.4.  t  
3. t c
     c chuy      
ng EMP (Embden-Mayerhof-p tc chuyn

2C
6
H
12
O
6
+ 3O
2

6
H
8
O
7
+ 4H
2
O
Glucose Acid citric

 i vi sinh v  sn xut acid citric: Aspergillus niger, Aspergillus
clavarus, Penicillium citrinum, Mucor piriformis 
Nhng chng Aspergillus cho kt qu cao nht.
3.4.2.  

2C
6
H
12
O
6
+ 3O
2

6
H
8
O
7
+ 4H
2
O
 ca s chuy c biu di
ng C
6
H
12
O
6
thu ph

3
COCOOH)
CH
3
COCOOH + CO
2

2
COCOOH
Acid oxaloacetic
HOOCCH
2
COCOOH + CH
3

2
COOHCOHCOOHCH
2
COOH
Acid citric
3.5. u t ng n n xut acid citric
3.5.1. Vi sinh vt
Vi sinh vt n la
chn vi sinh v 
ng hp acid citric xy ra mt ph o ra
acid cao, nu kh o ra acid ca vi sinh th sn xut ra
mt ng lc; mt ph  u

n vi sinh vt s ch c ch kh 
chng vi sinh vt s dng cn pho ra nhng acid h

u s 
n chng ca acid citric.
3.5.2. u
   p cho sn xu     u cha nhiu
: mt rt s  t
N axit citric cao nhc nn  
u cao (15 - 20% w/v), n ng cho vi sinh vt
s d    ng h       u phi
7
n nhiu tp, cht kim loi nc bit n
gii hn phi loi b.
3.5.3. ng
Nm mc cng, tinh bu king nm
mc ch  n sinh khi to ra r     ng h 
ng nhing 4 - ,
b  ng hp axit
citric xy ra mnh.
3.5.4. ng
 t 3 - ng nm mc s
d ng bng ph thu
nguu s dng NH
4
NO
3
u s dng NH
4
Cl
 t
pH gim my cn phu chnh pH cp.
3.5.5. S 

n mng ly cn phi cung c
cho qu 
ng.
3.6. u t quyn xut acid citric
3.6.1. 
Aspergillus niger
        - 140; NH
4
NO
3
- 2,23;
KH
2
PO
4
- 1; MgSO
4
.7H
2
O - 0,23.
3.6.2. 

4
Cl 1,
4



 
 - 3,5.

 


citric.
3.6.3. 






8
3.6.4. 
N- 37
0

34 - 37
0
 - 32
0

citric


Trong thc t sn xung th
men.
Nhi  37
0
C. Th
c to axit citric b 

men khong 7  
3.6.5. Th
Thi gian t     n xu   t t   thu 
chc s dc cng, h th
u ki
 mt, th-
trong khi  i gian  ngu khong t (5-
n th thuc
mnh m c s d m b m u,
nhi   ng m   ng  , v ht carob hoc sung).
ng h tc t trong khong 10


9

acid citric

acid







4.1. acid citric
4.1.1.

       - 4% trong      
35 - 38

0
            
-


- 0,2 at, t
0
= 34  35
0
-
- 10 m
3
- 30 - 100 m
3
/h.
4.1.

-
-
men.
  

-

4.1.

-.
- 34
0
acid- 32

0
C.

3

3
).
- 1000 m
3
/h.

10
4.1.
  
-
men.
-
- 65
0



4.1.
acid citric, acid gluconic, acid

 acid              


- 7,5.


C
6
H
8
O
7
+ 3Ca(OH)
2

3
(C
6
H
5
O
7
)
2

2
O
Acid citric Canxi citrat
2C
6
H
12
O
7
+ Ca(OH)
2


6
H
11
O
7
)
2
+ 2H
2
O
Acid gluconic Canxi gluconat
C
2
H
2
O
4
+ Ca(OH)
2

2
O
4

2
O
Acid oxalic Canxi oxalat



4.1.itrat

2
SO
4

- 0,5 m3/acid
acid
cacid citric trong citra

0
 cho H
2
SO
4
- 

2
SO
4
/1kg aciditrat)- 
Ca
3
(C
6
H
5
O
7
)

2
+ 3H
2
SO
4

6
H
8
O
7
+ 3CaSO
4
acidacid
   
acidacid


11
4.1.acid
- 1,26
- 1,%.
4.1.acid citric
Khi       - 37
0
        acid
citri- 10
0



 

0
C).
4.2. Sn xut acid citric bng nm mc Aspergillus niger


4: Aspergilus niger

4.2.1. 
Bng 1: ng s dn
xut acid citric bng nm mc Aspergillus niger.
n
Kho
 chung
Sucrose hoc glucose
125-225 kgm
-3

180 kgm
-3

NH
4
NO
3
(hoc mui NH
4
+


0.5  3.5 kgm
-3

1.5 kgm
-3

KH
2
PO
4

0.5  2 kgm
-3

0.5 kgm
-3

MgSO
4.
7H
2
O
0.1  2.0 kgm
-3

0.25 kgm
-3

Fe
2+

2 - 1300 mgm
-3

< 200 mgm
-3

Zn
2+
0-2900 mgm
-3

200-1500 mgm
-3

Cu
2+
1-10200 mgm
-3

200-1500 mgm
-3

Mn
2+
0-46 mgm
-3

<2 mgm
-3

12
4.2.3. 
Thc hi  u ki   chuy   t ging g  
ng Agar.
Sau 3 - y  nhi 30
0
 c thu hoy
ng tinh b khong 10
11
 cho 1cm
3
ng.
Sinh kh c truyn trc tip t n 10 - 20 cm
3

to ging dng ht cha 1 - 5.10
5
hng. Ging dng ht
c ti   p vi n
5 - 10% (v/v).
Nhi n men trong khong 28 - 35
0
C
ng cc khng ch  tr 2,2 - 2,6 bng vic b sung kim
NH
3

Vi t sinh acid citric p nm mc s u

,3 - 0,5 kgm
-3
h
-1

pH th nh
p s hn ch sinh tng hp acid citric
 c khng ch bng vic b t
4.2.4. Thu h acid citric
Thu hi acid citric  c thc hi
 Kt tinh trc tic dch lc
 Kt ta  dng tetrahydrate citrat canxi
 Chit pha lng
Ph bin nht ta citrat
Sinh khi nc l thng lc.
c kt ta t dch lc b sung canxi hydroxit
  nhi 70
0
C, sn phm ch y 

  nhi  90
0
C, sn phm ch yu l    
tetrahydrat tinh th
ng acid citric i trong dch l c kt ta mt ln na
b  5,8
Phn kt tc lc tip bng mc n dch lc loi b.
Kt ta  dng dicancium hydro citrate s d
t p theo.
Vic ra phn kt ta nhn loi b p chng, protein

thu  sinh khi nm.
13
 sau khi r ng thn hp
cha dihydrat sulphate canxi.
Dch lc x ng than hoi,
c loi b bi ci cation mu.
Dung di n 700 kgm
-3
 thng kt
ng  nhi 35
0
C hoc 62
0
C.
Tinh th acid citric  thng sy tng
n
u s d
0
n 2 s d nhi
20
0
C v  - 40%.
Khong 20% dch g     c ra thit b, lo  
chuyc v c x ng canci hydroxit. Phi ca dch gc
c lo b phn kt tinh


5: Tinh th acid citric

t pha lng, acid  d

dng hn hp trilauryl amine, n-c C11 isoparafin.
Dch chic gia nhia bc theo chio sn phm
c x ng ct than hoc
t t.








Tinh thể acid citric
14

6:  n xut acid citric bng nm Aspergillus niger
D

Mui citrat canxi
t ta

c
Tinh ch
Kt tinh
Ca(OH)
2
,
CaCO
3
10%

Acid citric
Nm mc
Tinh bt
H
2
SO
4

R ng
15
5
ng dng ca acid citric trong thc php

5.1. Trong thc phm
Vt cht ph gia thc phm, acid citric v,
cht bo qun thc ph uc bic gi
s E3305.
Mui Citrat ca nhiu kim lo    vn chuy   t
n ca ch.
c s d se, loi thuc
bc s dng trong truy
n c gi t ch

p
m ca  hiu ch pH ca cht
ty rc phm.  o phc vi nhiu kim lo
t ty ra. B
loc cc cht ty ra to nhiu bt
y sc.
  sn xu 

mc bi ra khi phc Citrat. Acid citric c
 sinh hc ph ch ng
d
Acid citric  t trong nh  t cn thi    ng hp
t cht d  ging Axeton
peroxit, nhy vi nhi mt s c nu bn mua mt s ng
ln acid citric bn s b li ng b.
  
i nh  i nghin thay
n bng mm hn ch s m c
b 
ng hp thiu thuc con nghin s  c chanh
vt thay gim.

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6
Kt lun ngh

5.1. Kt lun
Nhn thc tm quan tru ng dng ci
sn xup.
Sn xut acid citric t nm mc Aspergillus niger tn d c ngun
u, r tin, d kim li thu hi sinh khi cao.
5.2. Kin ngh
C u v acid citric 
m ru ngun vi sinh vt trong sn xut acid citric.
n xu thc hin xut tng
i hc An Giang.

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u tham kho

Nguyng (2002), Vi sinh vật trong công nghiệp (Công nghệ vi sinh
tập 2),  H t bn i hc Qu
ph H 
  (2005), Công nghệ lên men ứng dụng trong công nghệ thực phẩm,
 H : t bn i hc Qu H
Minh.
Nguyn Th  (2009), u kh Aspergillus niger
pectinmethylesterase     , Lu  t nghip K 
 Thc phm, c ng dng,
i hc C
Bảng dữ liệu an toàn acid citric (2012), m, Vin V sinh y t
 H 