I HC QUC GIA THNH PH H CH MINH

TRNG I HC KHOA HC T NHIN

CNG NGH SINH HC

VI SINH VT
Ging vin: TS. Nguyn Th Bch Hu
ti: Cng ngh sinh hc ca acid citric sn xut
Thnh vin_Nhm 4:
Nguyn Th Xun Diu

1118061

V ng Phng Lan

1118217

Hong Th Ca Ly

1118247

V Th Khnh Ngn

1118280

Nguyn Tn Anh Nhi

1118314

20/10/2014

Mc lc
I HC QUC GIA THNH PH H CH MINH ........................................................................................... 1
I.

Axit citric ................................................................................................................................................ 3

1)

Gii thiu .............................................................................................................................................. 3

2)

Thng tin tng qut ............................................................................................................................... 3

- Tn theo IUPAC: 2-hydroxypropane-1,2,3-tricarboxylic acid ................................................................... 3

3)

Tnh cht ............................................................................................................................................... 3

4)

Lch s tm ra ........................................................................................................................................ 4

5)

Sn xut................................................................................................................................................. 4

6)

ng dng .............................................................................................................................................. 4

II.

Aspergillus niger .................................................................................................................................. 5

1)

Gii thiu A. niger ............................................................................................................................ 5

2)

C s l thuyt ca qu trnh ln men acid citric .............................................................................. 5

3)

Mi trng ............................................................................................................................................ 6
Sn xut acid citric.......................................................................................................................... 9

III.

Cc phng php sn xut acid citric ............................................................................................... 9

1)
a.

Phng php ln men b mt ........................................................................................................ 9

b.

Phng php ln men chm......................................................................................................... 11

2) Cng ngh sn xut acid citric bng phng php ln men chm s dng nm mc Aspergillus
niger ........................................................................................................................................................ 12
3)
IV.
V.

Cc yu t quyt nh n qu trnh sn xut acid citric ................................................................ 14

Tnh hnh sn xut acid citric hin nay ....................................................................................... 15
Hng tng lai trong sn xut acid citric ..................................................................................... 15

I.

Axit citric

1) Gii thiu

Axt citric l mt axt hu c thuc loi yu v n thng c tm thy trong cc loi

tri cy thuc h cam qut. N l cht bo qun thc phm t nhin v thng c thm
vo thc n v ung lm v chua. lnh vc ha sinh th axt citric ng mt vai
tr trung gian v cng quan trng trong qu trnh trao i cht xy ra trong cc vt th
sng.
Ngoi ra axt citric cn ng vai tr nh l mt cht ty ra, an ton i vi mi
trng v ng thi l tc nhn chng oxy ha. Axt citric c mt trong nhiu loi tri
cy v rau qu nhng trong tri chanh th hm lng ca n c tm thy nhiu nht,
theo c tnh axt citric chim khong 8% khi lng kh ca tri chanh.

2) Thng tin tng qut

- Tn theo IUPAC: 2-hydroxypropane-1,2,3-tricarboxylic acid
-

Tn thng thng: axit chanh

Cng thc phn t: C6H8O7
Cng thc cu to:

C dng: tinh th mu trng

Khi lng phn t: 192.13 g/mol

3) Tnh cht

Tnh axt ca axit citric l do nh hng ca nhm carboxyl -COOH, mi nhm carboxyl
c th cho i mt proton to thnh ion citrat. Cc mui citrat dng lm dung dch m
rt tt hn ch s thay i pH ca cc dung dch axt.
Cc ion citrat kt hp vi cc ion kim loi to thnh mui, ph bin nht l mui canxi
citrat dng lm cht bo qun v gi v cho thc phm. Bn cnh ion citrat c th kt
hp vi cc ion kim loi to thnh cc phc dng lm cht bo qun v lm mm nc.
nhit phng th axt citric tn ti dng tinh th mu trng dng bt hoc dng
khan hay l dng monohydrat c cha mt phn t nc trong mi phn t ca axt citric.
Dng khan thu c khi axt citric kt tinh trong nc nng, tri li dng monohydrat li
kt tinh trong nc lnh. nhit trn 74oC dng monohydrat s chuyn sang dng
khan.

V mt ha hc th axt citric cng c tnh cht tng t nh cc axt carboxylic khc.

Khi nhit trn 175oC th n phn hy to thnh CO2 v nc.

4) Lch s tm ra

Vo th k th 8 nh gi kim thut Jabir Ibn Hayyan ngi Iran pht hin ra axt citric.
Cc hc gi chu u thi trung c cng bit v axt t nhin trong chanh, nhng kin
thc s b v axt ny cng c ghi nhn vo th k XIII. Axt Citric c nh ha
hc ngi Thy S tch c vo nm 1784, ng kt tinh c axt citric t nc
chanh p. Nm 1860 ngnh cng nghip nc p tri cy ca a cng trnh sn
xut axt citric vo hot ng.
Nm 1893 C. Wehmer pht hin ra rng nm mc (Penicillium) cng c th to nn
axt citric t ng. Sn xut axt citric theo kiu vi sinh ny khng c a vo sn
xut cng nghip cho n th chin th I, do cc xut khu nc hoa qu ca bc b.
Vo nm 1917 nh ha hc thc phm James Currie ngi M pht hin ra rng nm
mc hnh si (Aspergillus niger) c th dng sn xut axt citric rt hiu qu. Hai nm
sau tp on dc phm Pfizer ng dng k thut ny vo sn xut axt citric theo qui
m cng nghip.

5) Sn xut

K thut m ngy nay ngi ta vn dng trong cng nghip sn xut axt citric l nui
nm si trn ng n, sau lc nm mc ra khi dung dch v axt citric c tch
bng cch cho kt ta vi nc vi to thnh canxi citrat, sau kt ta c x l bng
axt sulfuric.
Ngoi ra axt citric cn c tch t sn phm ln men ca nc lo bng cch dng mt
dung dch hydrocacbon ca mt baz hu c Trilaurylamin chit. Sau tch dung
dch hu c bng nc.

6) ng dng

Vi vai tr l mt cht ph gia thc phm, axt citric c dng lm gia v, cht bo qun
thc phm v ung, c bit l nc gii kht, n mang m s E330. Mui citrat ca
nhiu kim loi c dng vn chuyn cc khong cht trong cc thnh phn ca cht
n king vo c th. Tnh cht m ca cc phc citrat c dng hiu chnh pH
ca cht ty ra v dc phm.

Citric axt c kh nng to phc vi nhiu kim loi c tc dng tch cc trong x phng
v cht ty ra. Bng cch phc ha cc kim loi trong nc cng, cc phc ny cho
php cc cht ty ra to nhiu bt hn v ty sch hn m khng cn lm mm nc
trc. Bn cnh axt citric cn dng sn xut cc cht trao i ion dng lm
mm nc bng cch tch ion kim loi ra khi phc citrat.

Axt citric c dng trong cng ngh sinh hc v cng nghip dc phm lm sch
ng dn thay v phi dng axt nitric.

Citric axt l mt trong nhng ha cht cn thit cho qu trnh tng hp Hexametylen
triperoxit diamin (HMDT) l mt cht d pht n ging Axeton peroxit, nhy vi nhit v
ma st. mt s nc nu bn mua mt s lng ln axt citric bn s b lit k vo s
en ca cc m mu khng b.

Axt citric cng c cho vo thnh phn ca kem gi cc git cht bo tch bit.
Ngoi ra n cng c thm vo nc p chanh ti.

Citric axit c coi l an ton s dng cho thc phm cc quc gia trn th gii. N l
mt thnh phn t nhin c mt hu ht cc vt th sng, lng d axt citric s b
chuyn ha v o thi khi c th.

iu th v l mc d axt citric c mt khp ni trong c th nhng vn c mt vi

trng hp mn cm vi axt citric. Tuy nhin nhng trng hp ny rt him v ngi
ta thng gi l phn ng gi v ca c th. Axt citric kh c th lm kch thch da v
mt do nn mc o bo h khi tip xc vi axt ny.

II. Aspergillus niger

1) Gii thiu A. niger
C nhiu loi nm mc c kh nng oxy ha gluxit thnh axit citric nh Rhizopus,
Aspergillus, Penicillium. Trong quan trng nht l Aspergillus niger.
Aspergillus niger l mt loi nm si v mt trong nhng loi ph bin nht ca chi
Aspergillus c nhiu cng dng trong cng ngh sinh hc, nhit ti u 35 370C,
pH t 1,4 - 9. N gy ra mt cn bnh c gi l nm mc en trn mt s loi tri cy
v rau qu nh nho, hnh ty, u phng, v l mt cht gy nhim ph bin ca thc
phm. Aspergillus niger l vi sinh vt hiu kh pht trin trn cc cht hu c, do n
c th c tm thy gn nh khp mi ni trong mi trng c cha t. Ngoi ra, n
c tm thy trong cht thi, nguyn liu thc vt phn hy v phn hu c trong mi
trng ngoi tri. Mi trng trong nh to ra mt mi trng sng tt cho s pht trin
ca nm mc. Chng hn nh trong cc lnh vc h gia nh thng thng, trn cc bc
tng trong phng tm hoc cc khu vc khu vc khc c m cao.
A.niger c nui sn xut nhiu cht trong cng nghip, trong c axit citric
2) C s l thuyt ca qu trnh ln men acid citric

Trong qu trnh ny glucose c chuyn ho thnh axit pyruvic theo con ng EMP
(Embden-Mayerhof-Parnas), sau axit pyruvic tip tc chuyn ho thnh axit citric.
Phng trnh tng qut ca qu trnh ny nh sau:
2C6H12O6 + 3 O2 6C6H8O7 + 4H2O

C ch ca s chuyn ho ny c th c biu din nh sau:

ng C6H12O6 thu phn thnh acid pyruvic (CH3COCOOH)
CH3COCOOH + CO2 HOOCCH2COCOOH

(Acid oxaloacetic)
HOOCCH2COCOOH + CH3COOH CH2COOHCOHCOOHCH2COOH
(Acid citric)
3) Mi trng
a s cc quy trnh cng nghip hin nay s dng ngun c cht l r ng. Nhng d c s
dng ngun c cht nh th no th ngi ta vn cn phi ch y n hai yu t quan trng l gi
thnh v cc yu t cn chun b cung cp iu kin thun li nht cho vi sinh vt ln men.
Ngun c cht c bn c dng trong cc quy trnh ln men ni l mt r ng ma hoc mt r
c ci ng. Cn i vi cc quy trnh ln men ni th ngun c cht c th c s dng
khng ch l mt r ng m cn l nhng c cht c tinh khit cao hn nh glucose tinh
khit, ng th hoc ng tinh, nc p ma v c ci ng c c. S dng cc ngun c
cht tinh khit nh trn c th gip lm nng nng sut hoc gip gim thi gian nui cy.
Di y l bn nng cc cht trong nui cy Aspergillus niger nhm ln men acid citric:
Thnh phn

Khong gi tr

Gi tr chung

Sucrose/Glucose

125-225 kg/m3

180 kg/m3

NH4NO3 (hoc mui NH4+ khc)

0.5-3.5 kg/m3

1.5 kg/m3

KH2PO4

0.5-2.0 kg/m3

0.5 kg/m3

MgSO4.7H2O

0.1-2.0 kg/m3

0.25 kg/m3

Fe++

2-1300 mg/m3

<200 mg/m3

Zn++

0-2900 mg/m3

200-1500 mg/m3

Cu++

1-10200 mg/m3

200-1500 mg/m3

Mn++

0-46 mg/m3

<2 mg/m3

1. Mt r
y l ngun c cht ph bin hin nay, c cht lng rt a dng ty theo ngun. V dng
sn xut acid citric th ta cn dng nhng ngun mt r c cht lng cao. Hai loi mt r ny c
thnh phn khng ging nhau, do ngi ta s chn mt trong hai cho quy trnh ca mnh.
Cng c nhng trng hp ngi ta trn c hai loi mt r vi nhau, nhm thu c nhng
ngun dinh dng t c 2 ngun. Cht lng lng ca ma hoc c ci ph thuc vo cc yu t
canh tc nh: iu kin t trng v kh hu, hnh thc bn phn, phng php gieo trng, thi
gian v iu kin bo qun, k thut sn xut, dng c k thut, quy trnh k thut,

R ng ma

R ma c khong 65-80% lng cht kho v 20-25% l nc. Thnh phn cc cht dinh dng
ca n c lit k trong bng sau:
ng tng s (%)------------------------------------- 48 56
Cht hu c khc(%) ---------------------------------- 9 12
Protein(%)----------------------------------------------- 2 4
K(%) ----------------------------------------------------- 1.5 5
Ca(%) ---------------------------------------------------- 0.4 0.8
Mg(%) --------------------------------------------------- 0.06
P(%) ------------------------------------------------------ 0.6 2
Biotin (vitamin H) mg/kg ----------------------------- 1 3
Axit pantothenic mg/kg ------------------------------- 15 55
Inositol mg/kg ------------------------------------------ 2500 6000
Thiamin mg/kg ----------------------------------------- 1.8
Trong ng tng s bao gm sucrose (44-54%), ng chuyn ha (0.4-1.5%), raffinose (0.52%), kestose v neokestose (0.6-1.6%), arabinose, xylose v mannose (0.5-1.5%). Trong ,
raffinose l mt thnh phn t nhin ca ng ma, trong khi kestose l sn phn ca cc chng
vi sinh trong qu trnh x l. Tt c cc ng trn (tr sucrose) u c xem l c cht hu c
khng nito trong r ng.Cc sn phm ha v nhit phn hy ca ng (melanoidine v
caramel) v cc acid hu c cng thuc nhm ny.Caramel cha ng anhydride v cc cht c
mu, cn melanoidine l sn phm ca mt phn ng gia ng gim v acid amin.
Ngoi ra, trong r ng cn cha cc acid hu c khng bay hi v bay hi, v nu lng acid
khng bay hi cao hn 1% th thng s c mu rt m v khng c dng cho quy trnh ln
men acid citric.
Cc cht cha nito trong r ma gm ch yu l betaine (c trong ma nhng khng c vi sinh
vt s dng lm ngun N), sau l acid amin, protein v cc cht vi lng nh ammonium
nitrate v amid.
Ngoi cc yu t trn th mt yu t cng rt quan trng nh gi cht lng r ma nhm t
nng sut cao trong sn xut acid citric l lng cc nguyn t vi lng trong mi loi r.
pH ca mt r ph thuc vo cng ngh chit, v pH c cho l ti u ln men acid citric l
trung tnh hoc kim nh. Nhm t c gim pH cn thit trong quy trnh sn xut acid
citric, ta cn mt r vi kh nng m thp.

R dng c ci

R ng c ci c thnh phn kh khc bit so vi r ng ma: cha t sucrose hn v nhiu

ng chuyn ha hn, cha t nito v raffinose, mu sc m hn v kh nng m cng thp
hn.

C hai loi r ngoi nhng thnh phn chnh cn cha cc yu t vi lng, d rt t nhng li l
thnh phn quyt nh xem r c iu kin a vo s dng sn xut acid citric hay khng.
Cc cht ny bao gm thuc tr su, tr nm v tr c (t qu trnh canh tc) v c cc cht ph
bt (t qu trnh sn xut ng), l nhng cht c tnh c v c nh hng tiu cc n cht
lng mt r.Thng th mt r ma c dng ph bin hn trong sn xut acid citric, c bit l
trong ln men chm, khi m cht lng ca mt r tr nn cng quan trng hn.
Cc vi sinh vt cng l nhn t nh hng n cht lng, c khi lng vi sinh trong mt r ln
n hn 10,000/g mt r (Bacillus, cc loi thuc Candida, Penicillium d rt him,
Aspergillus, v cc loi khc). Ngoi ra x l mt r ta cn phi loi mt s kim loi nng
bng nhiu phng php nh s dng kali ferrocyanide v EDTA (v mt s cht khc). Tuy
nhin cng cn lu gia li nhng cht cn thit cho Aspergillus niger s dng trong qu trnh
ln men.
2. Sucrose:
ng tinh luyn c xem nh l ngun sucrose nguyn cht m A.niger c kh nng s dng
rt tt.Nng ti u ca sucrose l vo khong 15% - 22%, v dng nhin ta cn thm vo
cc ngun dinh dng khc nh NH4NO3, KH2PO4, MgSO4, ng thi cn acid ha bng acid
sulfuric n khi t pH 2.6-3.0.
3. Syrup:
Xi-r ng cng c th c s dng lm c cht cho ln men acid citric chm.u im ni tri
nht ca ngun c cht ny l tinh khit ca n.Tuy nhin syrup li rt d gim cht lng
nhanh chng trong qu trnh vn chuyn v lu tr.V th ngun c cht ny ch c th s dng
trong nhng khong thi gian sn xut ng t cao im, v nh my ng phi rt gn vi
nh my sn xut acid citric.
i vi c cht ny, ta ha tan vo nc n nng 15-20%, thm vo cc cht dinh dng
cn thit (NH4NO3, KH2PO4, MgSO4 v (NH4)2C2O4), acid ha v sau tin hnh hp kh
trng.
4. Tinh bt
Sn xut acid citric bng ngun c cht l tinh bt ng, la m, khoai ty, tng i ph bin.
Ngun c cht ny c tinh khit v phng thc thy gii (bng acid hoc bng enzyme, hoc
kt hp c hai) kh ph hp cho vic sn xut acid citric.
S dng phng thc thy gii ca tinh bt bng enzyme cng c th s dng cho ngun c cht
l syrup dextrose.Quy trnh ny thm ch c p dng vo quy m cng nghip. Tuy nhin
cn phi ch n lng kim loi nng trong dch ln men v tin hnh loi b chng nu chng
vt ngng cho php.

5. Alkan
Vi gi thnh r v c nhiu chng vi sinh vt s dng, alkan lm thay i mnh m
phng thc sn xut acid citric trong nhng thp nin 1960 1970. V d in hnh l quy trnh
s dng nm Candida lipolytica. Hin nay alkan khng cn c s dng ph bin nh trc do
vn thu hi gp kh khn (do s to thnh acid isocitric).
6. Du & cht bo
Hin nay du c s dng lm ngun carbon chnh sn xut acid citric tng t nh alkan
trc y. Trong mt v d c th: du c c chng t bin ca Candida lipolytica c th ln
men to acid citric vi hiu sut ln n 145%. Ngoi ra du cng c thm vo mi trng ln
men bng Aspergillus niger vi nng thp, thm ch c trng hp l ngun carbon duy nht.
Ngi ta thy rng y l ngun carbon c th cho nng sut cao trong vic to acid citric.Du
v cht bo c th thay th alkan trong vi quy trnh, tuy nhin gi thnh th cha hn vn cn
mc thp.
III.
Sn xut acid citric
1) Cc phng php sn xut acid citric:
Qu trnh tng hp sn xut acid citric bng cng ngh ln men l phng php c gi tr
kinh t nht v c s dng rng ri trn ton th gii. Hn 90% lng acid citric trn th gii
c sn xut bng cng ngh ln men, v chng c nhng u im sau:
(1)
(2)
(3)
(4)

quy trnh n gin v n nh

nh my thng t phc tp v cn t h thng iu khin phc tp
yu cu k thut cn thit t
tiu th nng lng t hn v nhng s c mt in thng xuyn khng gy nh
hng nghim trng n hot ng ca nh my sn xut.

Sn xut acid citric bng cng ngh ln men gm 3 giai on:

(1) chun b v cy nguyn liu th (inoculation)
(2) ln men
(3) thu hi sn phm.
Trong sn xut acid citric theo quy m cng nghip, thng thng ngi ta s dng 3 phng
php ln men: ln men chm, ln men b mt v ln men bn rn (solid-state) hay cn gi l qu
trnh Koji.
a.

Phng php ln men b mt

Phng php ln men b mt ng dng nhiu vo nhng nm u ca th k XX, theo cc vi

sinh vt pht trin hn trn b mt mi trng nm gia pha rn v pha kh hoc nm gia pha
lng v pha kh. Nh vy, phng php ln men b mt c th s dng hai loi mi trng: mi
trng bn rn, cn gi l mi trng xp v mi trng lng.

+ Ln men b mt vi mi trng bn rn
Phng php ny t c p dng v hiu sut thu khng cao v kh phc tp khu chit, tch
acid citric. Trc kia ngi ta dng mi trng cm m, cm go c trn khong 15 - 25% tru
tng xp ca mi trng. Ngoi ra, trong mt s c s ngi ta cn dng mi trng khc
t sn, khoai ty, bp Mi trng lm nguyn liu phi c nghin nh n kch thc va
phi. Sau khi nghin xong mi trng phi c lm m n m khong 60 - 65% v em
hp thanh trng bng hi nng. Mi trng lm xong phi c lm ngui v chun b cho qu
trnh nui cy. Trc ta phi chun b ging. Qu trnh ny cn gi l giai on thu nhn bo
t ging.
Phng php thu nhn bo t ging: C 2 phng php thu nhn:
Phng php th nht: Chun b 3 - 4 bnh tam gic dung tch 150 ml, cho vo khong 50g
cm c m 60%. Hp thanh trng v ngui.
T ng ging Aspergillus niger gc, ta chuyn ton b khun lc bng cch cho vo mi ng
nghim 10ml nc v trng, khuy u cho bo t ging trong ng nghim ha trn trong nc.
Bng phng php v trng chuyn ton b sang cc bnh tam gic chun b mi trng sn.
Lc u cho mi trng trn u bo t. Nui chng trong t m c nhit 30 - 37oC, trong
thi gian 3 ngy. Khi thy trong bnh tam gic ton l bo t mu en l kt thc giai on nui
cy.
Tip , cng mi trng nh trn nhng c chun b vo khay nhm hoc inox. Ti u khi
cho 10% mi trng ging t bnh tam gic nui trn vo. Chiu dy khi trn khong t 3
- 5cm. trong phng c nhit n nh l 32oC. Nui trong khong 3 - 4 ngy khi bo t
mu en xut hin kn b mt mi trng, em sy nhit < 40oC ta thu c ging bo t
sn sng cho qu trnh sn xut i tr.
Phng php th 2: Ngi ta nui trong mi trng lng ging Aspergillus niger. Mi trng
thu nhn bo t c thnh phn nh sau:
Dung dch nc malt c nng cht kh 3 - 5%.
NH4Cl:0,25g
KH2PO4 1,25g
MgSO4 0,25g
FeSO4 0,0125g
Mi trng chun b xong c a vo bnh c dung tch 2 - 3lt vi chiu cao ca dung
dch trong cc bnh l 1cm, em hp thanh trng 1 at trong 30pht. Sau chuyn ging t
ng nghim ging vo. Tin hnh nui nhit 32oC trong thi gian 4 ngy hoc lu hn cho
n khi trn b mt xut hin vng nm si, lc u l mu trng sau l en cha ton bo t.
Ngi ta thu bo t ny em sy nhit < 40oC v dng n nh bo t ging cp 1. sn
xut bo t ging cp 2,3, ngi ta thc hin nh qu trnh nui nm si trn khay trnh by
trn.
Cch thc thc hin qu trnh ln men b mt:
Trn cm vi nc theo t l 1:1. Hp thanh trng 1at trong 30 pht v ti u ra khay, sau khi
lm ngui s c trn ging vi t l 0,3 - 0,5%. Chiu dy ca khi cm + bo t ging

khong 3 - 5cm. Tin hnh nui nhit n nh l 30 320C trong 4 - 5 ngy. Thi gian ln
men kt thc khi bo t nm si mi bt u xut hin nhiu nhng cha hon ton chuyn qua
mu en. Phng php nui cy b mt thu nhn acid citric trn mi trng bn rn hin nay
khng cn p dng cc nc chu u na. Tuy nhin, nhiu nc vn phng php ny vn
cn c p dng v thay cm bng bt khoai m. C 3 4kg bt khoai m ngi ta thu c
1kg acid citric.
+Ln men b mt vi mi trng lng
Thng thng ngi ta thay ng saccharose bng mt r. Mt r c x l trc khi lm mi
trng. X l mt r gm 3 vn cn gii quyt: -X l mu.
-X l h keo c trong mt r
-X l st nu mt r cha st
Mt r ng cn phi c x l mu v h keo bng cch cho dch qua than hat tnh hp
ph v pha long iu chnh h keo ca dch mt r. Nu trong mt r c Fe3+ ngi ta cho
vo mt r sau pha long K4{Fe(CN)6}.
Tip theo ta hiu chnh hm lng ng v pH thun li cho qu trnh ln men. Hm lng
ng c hiu chnh hm lng t 14 - 15% v khi lng.
Ngoi ra qu trnh ln men thun li cho vi sinh vt ngi ta b sung MgSO4, NH4NO3,
KH2PO4 v iu chnh pH t 2,2 - 6 ty thuc vo thnh phn cc cht b sung m iu chnh
pH cho hp l. Qu trnh chun b ging c trnh by mc trn.
Mi trng ln men phi c lc k v thanh trng, lm ngui, phn phi vo cc khay, nui
nhit 28 32OC trong thi gian t 48 - 72 gi. Qu trnh ln men trn b mt ca dch lng
nh lp vng dy trn l khun lc ca nm Aspergillus niger.
Acid citric s c thm thu qua mng t bo vo mi trng. Kt thc qu trnh ln men,
ngi ta ly phn dch ln men em i lc thu acid citric v tip tc ln men m mi.
b. Phng php ln men chm
Trong cng ngh sn xut acid citric theo phng php chm, ngi ta s dng mi trng ging
nh mi trng lng dng trong phng php ln men b mt.
Qu trnh c thc hin trong cc thit b ln men c cnh khuy v c h thng thi kh lin
tc. Ngi ta tin hnh ln men 28 320C trong thi gian 6 7 ngy.
Trong qa trnh ln men, ngi ta thng phi s dng CaCO3 iu chnh pH v acid citric
c to thnh s lm gim pH xung 1 1,5. Vic iu chnh ny cn c ngha l ngi ta
chuyn acid citric thnh xitrat canxi lng xung. Kt thc qu trnh ln men, ngi ta s dng
H2SO4 tch acid citric ra. Tin hnh c c v kt tinh acid citric.
Trong k thut sn xut ny, cng ngh sn xut cng nghip chnh cho axt citric, cc
m cy nm Aspergillus niger c nui trong mi trng cha sucroza hayglucoza sinh ra
axt citric. Ngun ng l nc ngm ng c c, nc r ng, tinh bt ng thy phn hay
cc dung dch ng r tin khc. Sau khi nm c lc ra khi dung dch c to thnh, axt
citric c c lp bng kt ta n vi vi ti (hydroxit canxi) to ra mui citrat canxi, t
axt citric c sinh ra bng x l mui ny vi axt sulfuric.

Ngoi ra axt citric cn c tch t sn phm ln men ca nc lo bng cch dng mt

dung dch hydrocacbon ca mt baz hu c Trilaurylamin chit. Sau tch dng dch hu
c bng nc.
So snh gia ln men chm v ln men b mt :
Mc d qu trnh sn xut b mt cn t nng lng hn, nhng rt nhiu nhc im
trong . Yu cu khng gian ln hn cho sn xut v c lp, yu cu s thng hi v v trng
cao hn. Mt trong nhng vn ln nht ca qu trnh sn xut l v trng. u im chnh ca
qu trnh ln men chm l: thi gian ln men ngn (6-7 ngy), m bo mc v trng cao hn
v kim sot cc thng s qu trnh, qu trnh hot ng n gin, yu cu khng gian nh hn,
qu trnh lp li v nng sut cao hn.
Schierholt so snh li nhun kinh t ca qu trnh ln men b mt v ln men chm trong
sn xut axit citric. Vi 300m3 v 150m3 ln men trong 9 ngy thu c nng sut 72 tn v 12
tn mi ngy. ng kt lun chi ph u t xy dng qu trnh ln men b mt cao hn so vi ln
men chm 2,5 ln. Tri ngc vi iu ny, cc chi ph v trang thit b l cao hn ng k qu
trnh ln men chm, v hn 60 phn trm nhng chi ph bao gm cc thnh phn phc tp cng
nh cc phn ng sinh hc v kim sot cng c tinh vi hn...
Tng chi ph u t cho qu trnh chm thp hn khong 25 phn trm cho dung lng
cao hn v thp hn 15 phn trm i vi dung lng nh hn so vi qu trnh ln men b mt.
Chi ph u t cho qu trnh chm ngc vi chi ph sn xut cao hn ng k so vi cc quy
trnh khc. c bit r rng l tiu th nng lng in cao, cao hn khong 30 phn trmso vi
yu cu ca qu trnh ln men b mt. Chi ph lao ng cc nc pht trin cao cho nnln
men b mt c s dng nhiu hn. Cc nc ni nhit nc lm mt vt qu 20C, thm
chi ph lm mt cho cc phn ng sinh hc xy ra do phi ci t h thng lm mt tng
hp cho qu trnh ln men chm. Qu trnh ln men chm nhy cm nu b gin on ngn hoc
s c trong thng kh, kt qu khng ch mt nng sut, m cn gy ra s c trn cc l tng
ng. Qu trnh ln men b mt, cc dung dch axit citric hoc dch ln men tp trung nhiu hn
so vi qu trnh ln men chm, b nh hng bi s bay hi cao hn qu trnh ln men chm.
Sn xut axit citric bi b mt rn hoc bng c lp t cam, qut,nc tri cy chim t l
khng ng k trn quy m th gii. Mc d c hai qu trnh sn xut vi chi ph thp, chng
c s dng ch yu cc nc c truyn thnglu i (, Hy Lp, Chu ).
2) Cng ngh sn xut acid citric bng phng php ln men chm s dng nm mc
Aspergillus niger
Cng ngh sn xut acid citric gm 8 giai on
-

Nui cy nm mc (nui cy trong phng th nghim v nhn ging trong sn xut)

Chun b dung dch r ng 3-4% trong nhng thng nui cy nhit 35-380C.
B sung dung dch cc cht dinh dng vo thng nui cy. Chuyn men ging t
phng th nghim vo theo t l 3g bo t kh/ 2-3 lit dung dch r. Sau m cnh

khuy v cung cp khng kh v trng (np khng kh v o trn sut qu trnh nhn
ging). Duy tr p sut trong thng 0.1-0.2at, nhit bng 34-350C v thi gian 2836h. Thi k u cho oxy vo ti lng 9-10m3/h, thi k cui (24-30h) l 90100m3/h.
Chun b dch ln men:

Trc ht phi dng hi cao p tit trng thit b v ng ng. R ng c pha thnh 2
loi nng : nng 3-4% nui cy mc ging v ln men ban u. Nng 25-28% b
sung trong qu trnh ln men, pha ch dch ln men, dng nc v trng trn vi dung dch
cc mui dinh dng v r ng ri khuy u. Mi trng 3-4% c pha ch trong thit b
ln men. Sau cho mc ging t thit b nui cy vo v tip tc khuy trn trong 30 pht.
-

Ln men:

Trong qu trnh ln men lng ng gim nhanh, b li dng dung dch r c nng 2528% b sung gin on vo dung dch ln men. Thi k u gi 33-34oC , khi to acid
mnh th gi nhit 31-32oC. Thi k u cung cp 100m3/h ( th tch thit b 50m3). Thi k
cui 800-1000m3/h.
-

Tch nm mc:

Kt thc qu trnh ln men bng cch kim tra mu. Nu hai mu kim tra cch nhau 4-6 gi m
c acid nh nhau th coi nh kt thc qu trnh ln men. Thi gian ln men c th ko di 510 ngy, ph thuc vo hot lc ca nm mc. Khi kt thc qu trnh ln men th un nng dch
ln men 60-650C v chuyn vo thng trung gian tch nm mc. Nm mc c tch trn
my lc chn khng.
-

To canxi citrate

Dung dch ln men l hn hp gm: acid gluconic, acid oxalic, ng khng ln men v cc
hp cht khong.
Tch acid citric bng cch cho lin kt vi cation Ca to mui t tan canxi citrate. Dung dch
ln men cho vo thit b trung ha v em un si. Sau m cch khuy v cho sa vi vo
trung ha. Qu trnh trung ha c kt thc khi pH =6.8 7.5.
Khi trung ha to thnh
C6H8O7 + 3Ca(OH)2 Ca3(C6H5O7)2 + 6H2O
Acid citric

canxi citrate

2C6H12O7 + Ca(OH)2 Ca(C6H11O7)2 + 2H2O

Acid gluconic

Canxi gluconat

C2H2O4 + Ca(OH)2 CaC2O4 + 2H2O

Acid oxalic

canxi oxalate

Dng thit b lc chn khng tch cc cht kt ta canxi citrate v canxi oxalate ri em sy kh.
-

Tch canxi citrate: Dng H2SO4 tch canxi citrate (trong thit b tch c cnh
khuy, ng phun hi v thot hi). u tin cho nc vo thit b 0.25-0.5m3/1 tn
acid citric cha trong citrate, m cnh khuy v cho cht kt ta vo. lm trong
acid citric, dng than hot tnh vi lng 2% so vi lng acid citric trong citrate.
Sau em un nng ln 600C v cho H2SO4 c t trng 1.8-1.84 vo (0.425l
H2SO4/1kg acid citric c trong citrate). Khuy u ri un si 10-15.

Ca3(C6H5O7)2 + 3H2SO4 2C6H8O7 + 3CaSO4

tch canxi oxalate khi c mt acid citric, s dng 1 lng d acid sulfuric, khi canxi
oxalate s kt ta vi thch cao c to thnh v lc trong dung dch ch cn acid citric.
tch dung dch acid citric khi kt ta c cha thch cao, canxi oxalate, than, cc hp cht sulfur
ca kim loi nng, chuyn ha hn hp vo lc chn khng, dung dch sau khi lc em sy.
-

Sy dung dch acid citric trong thit b sy chn khng:

+ Giai on u sy n t trng 1.24-1.26.

+ Giai on hai sy n t trng 1.32-1.36 tng ng vi nng 80%.
-

Kt tinh v sy kh acid citric:

Khi nhit ca dung dch t 35-370C th cho mm kt tinh (tinh th acid citric) vo kt tinh,
tip tc lm ngui 8-100C v cho khuy lin tc trong 30. Sau cho qua thit b ly tm tch
tinh th ri a i sy kh (dng thit b sy kiu bng ti, tc nhn sy l khng kh vi nhit
khng qu 350C).
3) Cc yu t quyt nh n qu trnh sn xut acid citric:
-Mi trng dinh dng bao gm ng, cc hp cht hu c, v c. nui cy Asp. niger s
dng mi trng c thnh phn (g/l):
Saccharose 140; NH4N3 -2.23; KH2PO4 1; MgSO4.7H2O 0.23.
-Mi trng ln men bao gm:
Nc 1000ml + ng 150g + NH4Cl 1.9g v b sung ZnSO4 lm tng kh nng tch ly acid
citric.
-pH ca mi trng:
+ nm mc pht trin tt gi pH = 6
+ ln men tt gi pH 3.4-3.5
+ iu chnh pH thng dng mt s acid nh HCl.
V iu kin mi trng nm pht trin v thu acid citric l khc nhau nn trong sn xut
phi chun b mi trng cho nm pht trin y , sau iu chnh mi trng thch hp
ln men citric.

-S thong kh: Tt c h si ca nm mc l loi hiu kh in hnh, rt cn oxy t do. Trong

sn xut c th dng qut gi v trng vo phng ln men hoc thi kh v trng vo dch ln
men.
-nh hng ca nhit : Nhit thch hp khong 31-370C. Sinh khi nm mc pht trin
mnh 34-370C. to ra nhiu acid, cn duy tr nhit 31-320C. to ra nhiu acid, cn
duy tr nhit 31-320C. Nhit thp hn th tch ly nhiu acid gluconic. Nhit cao hn th
vic to acid citric b km hm.
-Thi gian nui cy 7-10 ngy.
IV.

Tnh hnh sn xut acid citric hin nay

65% lng acid citric sn xut c s dng vo cng ngh thc phm v nc gii kht. Lng
acid citric c sn xut:
-

Nm 2004 l 1.4 triu tn

Nm 2005 l 1.6 triu tn

Trong Trung Quc l nc c lng sn xut cao nht, chim khong 50% sn lng ca
th gii. Trong khi sn lng ca c chu u v M chim khong 1/3 th gii.

V lng s dng th 3 khu vc trn cng l 3 khu vc c mc tiu th cao nht, chim tng
cng khong 65%-70% ca th gii.
V.

Hng tng lai trong sn xut acid citric

Hin nay acid citric khng cn c nhn nhiu s quan tm ca sinh hc phn t, mt phn l
do y l mt lnh vc tng i hon thin.

Hin nay, vi nhng hiu bit tch ly v cc c ch iu ha, vic ci bin qu trnh chuyn ha
c th thc hin c.
-

Cng ngh ln men m c th thay th bng cng ngh ln men bn lin tc nhm nng
cao nng sut v h gi thnh. Tuy nhin, iu ny s dn n s ph hy h si ca nm.
Nguyn nhn vn cha c gii p.

Khi nng NH4+ ni bo gim di mc 1mmol/g sinh khi kh, sinh tng hp acid
citric b ngng li, nm thay i hnh thi theo hng bt li => b sung nng thp
(30g/m3 ~ 2mmol/g sinh khi kh) trong qu trnh ln men. Nu iu ny c khng
nh, ch cn quy m pilot trong thi gian ln men di n c th dn n mt s thay
i ton din trong sn xut acid citric.

Kh nng c nh sinh khi nm bng vt liu v c hoc hu c v s dng trong thi

gian nhiu tun, hoc vi thng ang cn l mt thch thc.

Cng ngh thu hi truyn thng lm pht sinh mt s vn lin quan ti vic x l dch
thi (vi COD khong 20kg/m3) v cht thi rn (c th l khong 0.15kg sinh khi kh,
2kg thch cao cho mi kg acid citric khan). Mt s cng ngh c xut: thay th
nguyn liu r ng bng tinh bt thy phn nhm gim phc tp trong qu trnh thu
hi. C th thu hi qua 1 bc duy nht nu thu hi tricalcium citrat t dch ln men
lc v loi mu bng in ly gii, v acid citric c hp ph ln nha trao i anion yu
hoc ziolit s dng k thut sc k trao i ion pha rn chuyn ng. Sn phm sau c
th thu hi bng nc, acid long hoc s dng mng lng => hn ch lng cht thi rn
to ra => gim thiu chi ph x l.

Ti liu tham kho

http://www.authorstream.com/Presentation/chweetpia-1395191-citric-acid/
http://baigiang.violet.vn/present/show/entry_id/8212525
http://luanvan.net.vn/luan-van/quy-trinh-len-men-acid-citric-tu-aspergillus-niger-58815/
Cng ngh sinh hc tp 5 Cng ngh vi sinh v mi trng GS.TS. Phm Vn Ty TS. V
Nguyn Thnh
C s cng ngh sinh hc tp 4 Cng ngh vi sinh L Vn Hng, Nguyn Vn Cch.
Citric acid production - Marin Berovic, Matic Legisa