B CNG THNG

TRNG I HC CNG NGHIP TP.HCM

VIN CNG NGH SINH HC- THC PHM
........................

Bi tp:

TM HIU V PH GIA ACID GLUTAMIC

LP: DHTP9B-N1
GVHD: NGUYN TH HONG YN
SVTH: 13049821- ON TN HUY
M HP: 210502602

MC LC:

I. Lch s m chnh...............................................................................................1
II. Gii thiu v ph gia glutamic........................................................................2
2.1. Khi nim v tnh cht vt l........................................................................2
2.2. Tnh cht ha hc..........................................................................................3
2.3. Vai tr ca acid glutamic v natri glutamat..................................................6
III. Cc phng php sn xut m chnh .............................................................7
3.1. Phng php tng hp ha hc....................................................................7
3.2. Phng php thy phn protit.......................................................................8
3.3.Phng php ln men.....................................................................................9
3.4. Phng php kt hp....................................................................................10
IV. C ch to axit glutamic ca chng Micrococcus glutamicus.......................10
4.1 T ngun cacbon l sacharide theo chu k Embden- Mayerhaf....................10
4.2. Cc sn phm ca qu trnh ln men L-AG..................................................12
4.2.1. Sn phm chnh..........................................................................................12
4.2.2 Sn phm ph..............................................................................................12
4.2.3.S chch hng to sn phm chnh...........................................................14
V. K thut sn xut axit glutamic v bt ngt....................................................15
5.1. Nguyn liu....................................................................................................15
5.2. Chng vi sinh.................................................................................................15
VI. c tnh v liu lng s dng......................................................................17
VII. Bt ngt c lin quan g n Hi chng qun n Trung Hoa khng.........19
VIII. Lu khi ch bin........................................................................................20

TM HIU V PH GIA GLUTAMIC TRONG THC PHM

I. Lch s m chnh:
Vo nm 1860 nh khoa hc Ritthaussen Hamburg (c) xc nh thnh phn trong
protein ng vt, c bit l thnh phn cc axit amin, trong c mt axit amin vi tn
gi l axit glutamic v mui natri ca n l glutamat natri, tip sau l Woff, mt nh
ha hc thun ty xc nh s khc nhau ca cc axit amin v trng lng phn t v cu
trc cng nhng hng s v l ha tnh ca chng.
Lch s m chnh c th cm mc u tin l ngy chng thanh nin Tokyo c tn l
Ideka theo hc ti Vin i hc Tokyo tt nghip c nhn ha hc nm 1989. Sau ny khi
qua c tu nghip Ideka c lm vic vi Woff. Chnh trong khong thi gian qu
bu ny Ideka hc c cch nhn bit v tch tng axit amin ring r. Sau ny khi
ng tr li Nht Bn, trong ba n gia nh khi cc mn n c ngi v ca ng cho
thm rong bin th v ca thc n c sc hn ln, ngt hn, v c v tht hp dn. Sau
ti phng th nghim ring ca mnh, Ikeda tm hiu trong rong bin c cht no m
lm cho thc n m v tht.
Ti phng th nghim ring ca mnh, Kikunae Ikeda tm hiu rong bin c cht no m
lm cho thc n thm m v tht. ng khng ng cng trnh nhn bit hot cht trong
rong bin ca ng li m ng cho mt ngnh cng nghip hng mnh th k 20.
T nghin cu c bn Ikeda tnh c axit glutamic t rong bin Laminaria Japonica
ri chuyn thnh Natri glutamat. Ikeda gi bn hn vn lp mt cng ty sn xut
glutamat Natri m ng t tn cho thng phm ny l Ajinomoto theo ngha ting Nht
l tinh cht ca v ngon. Ngy 21 thng 4 nm 1909, Ikeda ng k bn quyn sng
ch s 9440 ti Anh quc vi nhan : sn xut cht to v. Thc ra ngi ta bit axit
glutamic trc khi bit mui Natri glutamat l mt cht iu v. Tn axit glutamic xut
pht t thut ng Gluten ca bt m. Tch gluten, thy phn n bng axit v cui cng
thu c mt lng ln axit amin, trong axit glutamic chim 80% lng cc axit
amin.
Nm 1920, b mt v cng ngh sn xut mononatri glutamat (MSG) cng c khm
ph. Ngi cnh tranh vi Ajinomoto li chnh l ngi lng ging chu khng l, l
cc doanh nghip Trung Quc. Bt u t nm 1920 n nm 1930, hng V Tinh (Vi
Tsin) m dn min Bc gi chch i l m chnh sn xut hng nm 200 tn, cn Nht
lc sn xut hng nm c 4000 tn. Khi Nht m cuc chin tranh xm lc Trung

Quc, cc nh sn xut m chnh ca Trung Quc b dp b.

Mi n nm 1968 cng ty Ajinomoto ca Nht Bn mi hon thin qu trnh sn xut
m chnh thng phm bng phng php tng hp da vo cht ch yu l acrylonitrile
(CH2=CH - CN). Khi , cng ty ny mi ch sn xut m chnh bng phng php tng
hp.
Ti thnh ph Thng Hi trong sut nhng nm u ca th k 20 ngnh cng nghip
sn xut m chnh pht trin kh nhanh v n tr thnh mt sn phm thng dng
vi hu ht ngi dn Chu . Mc d vy lc ny m chnh l mt sn phm kh t,
nm 1952: 1kg m chnh gi khong 3,5 la. Nm 1956 cc qui trnh ln men dng tinh
bt lm nguyn liu ban u pht trin mnh lm gim gi thnh m chnh ,sau nm
1964 ngi ta s dng r ng ma lm nguyn liu sn xut m chnh lm cho gi m
chnh tip tc gim, iu ny to tin cho vic sn xut m chnh trn qui m thng
mi, cho dn nm 1968 gi m chnh khong 0,9 la/1 kg.
Ngy nay, vic sn xut axit glutamic ri chuyn thnh MSG (monosodium glutamate m chnh) khng nh bui ban u. Ngi ta khng tch axit glutamic c sn trong t
nhin nh t gluten ca bt m, hoc t rong bin m dng cng ngh vi sinh. T tinh bt
(ch yu l tinh bt sn - cung cp hydratcacbon) vi ging vi sinh vt v ngun Nit
to thnh axit glutamic ri chuyn mononatri glutamat. Theo cc nh kinh t mi nm
Vit Nam tiu th lng m chnh khong 50 triu USD. Theo t China Post (10/3/1993 i Loan) hu nh cc hng m chnh i Loan chuyn ra nc ngoi sn xut, nu sn
xut i Loan th gi 1 tn phi chi t 1200 1300 USD, cn sn xut nc ngoi th
chi ph thp hn, khong 800 900 USD.
II. Gii thiu v ph gia axit glutamic
1. Khi nim v tnh cht vt l
Axit glutamic l mt trong nhng axit amin khng thit yu, c th con ngi c
th t tng hp, thuc loi axit amin c 2 gc carboxyl v 1 gc amin. INS 620, tn
ting anh l Glutamate Natri.(L(+)-). L nhng tinh th khng mu, t nc =247~249oC,
nhit thng hoa 200oC t tan trong nc, etanol; khng tan trong ete, axeton. Axit
L (+)- glutamic c v ngt ca tht cn axit D (+)- glutamic.
Cng thc cu to ca acid glutamic:
Natri Glutamat
ting

Anh

Monosodium glutamate monohydrate, vit tt l MSG. Cc tn IUPAC: 2-

aminopentanedioic acid, 1-aminopropane-1,3-dicarboxylic acid. Tn quc t v cng

ng chu u: INS 621, EEC 621 (thng gi l cht iu v 621). Bt ngt chnh l
cht iu v 621. Cng thc phn t l: C5H8NO4Na. Trng lng phn t: 187,13.
Cng thc cu to ca natri glutamat l:
M chnh l loi
bt trng hoc tinh
th hnh kim ng nh, kch thc tu theo iu kin khng ch khi kt tinh.M chnh
thun 99%, tinh th hnh khi 1 2 mm mu trong sut, d dng ho tan trong
nc, v khng ha tan trong cn ,thm, ngon, kch thch v gic. V ca MSG c
th nhn ra r nht trong khong pH = 6 8.
Mui MSG thng dng to v cho thc phm v nng MSG thng trong
khong 0,2 n 0,5%. C 3 loi MSG l dng L,D v LD-MSG nhng trong
ch c dng L-MSG l to nn hng v mnh nht . - Thun m chnh l t l %
glutamat natri trong sn phm, hin nay thng sn xut loi 80 99%.
+ Trng lng phn t 187.
+ Nhit nng chy 1950 C. + pH = 6,8 7,2.
+ ho tan: tan nhiu trong nc, nhit tng ho tan tng. 250 C ho tan
l 74,0 g/100ml nc; 600 C ho tan l 112,0 g/100ml nc; 800 C ho tan l
32 340 Be.
+ Dung dch 10% MSG trong sut, khng mu,gi tr pH khong 6,7 7,2.
2.Tnh cht ha hc:
Tnh hot quang:
C ch hot ng nh cc axit amin khc c 2 dng ng phn quang hc nh cc
amino axit khc v c 2 dng ng phn quang hc D v L c C bt i. ng phn
L c v thm ngon, ng phn D c v khng thm ngon nn hn ch trong sn
xut. Trn th gii hin nay ngoi vic xc nh hm lng glutamat natri cn xc
nh thm hm lng L-glutamic bng my o gc quay cc nh gi thm cht
lng, trong L200C=+25,16

Phn ng phn hy nhit cao

Nung Natri glutamat nhit cao >350C

nhit cao trn di 100C, axit glutamic trong dung dch nguyn cht
b mt nc v chuyn thnh axit hydroglutamic.
S mt mt axit glutamic trong dung dch nguyn cht khi un nng l rt
nhanh: sau 8 gi un si axit glutamic b mt n 50%.
Tc dng vi axit v c v andehyde formic (HCHO)

Tc dng ca pH:

S mt mt axit glutamic(%) cc pH khc nhau

Thi gian un
nng (gi)
pH=4,5

pH=6

pH= 7,5

8,7

6,1

5,12

10,1

9,0

6,8

12,3

12,1

8,4

18,4

15,6

10,3

24,1

19,0

12,7

pH c nh hng rt ln n s phn hu axit glutamic. pH= 4,5 axit

glutamic tn hao nhiu nht: sau 1 gi l 8,75%; sau 5 gi tng ln 24,1%.
Trong khi nu mi trng l trung tnh hay cc im ln cn(pH= 6,5-7) th
s mt mt gim c rt nhiu.
Phn ng mt nc:
Khi nhit cao hn 80C th s xy ra phn ng mt nc:

3. Vai tr ca acid glutamic v natri glutamat

Vai tr ca L-AG Trong nhng nm gn y, vic nghin cu sn xut axit
glutamic c y mnh nht. Cng ngy ta cng s dng nhiu axit glutamic trong
vic nng cao sc kho v iu tr mt s bnh ca con ngi.
Axit glutamic rt cn cho s sng, tuy l mt loi amino axit khng phi thuc loi
khng thay th nhng nhiu th nghim lm sng cho thy n l mt loi axit amin
ng vai tr quan trng trong qu trnh trao i cht ca ngi v ng vt, trong
vic xy dng protit, xy dng cc cu t ca t bo. Axit glutamic c th m
nhim chc nng tng hp nn n tham gia vo phn ng chuyn amin, gip cho c
th tiu ho nhm amin v tch NH3 ra khi c th t cc aminoaxit khc nh
alanin, lsin, cystein, prolin, oxyprolin v.v... . N chim phn ln thnh phn protit
v phn xm ca no, ng vai tr quan trng trong cc bin i sinh ho h thn
kinh trung ng, v vy trong y hc cn s dng axit glutamic trong trng hp suy
nhc h thn kinh nng, mi mt, mt tr nh, s u c NH3 vo c th, mt s
bnh v tim, bnh teo bp tht v. v... L-AG dng lm thuc cha cc bnh thn kinh
v tm thn, bnh chm pht trin tr c tr em, bnh bi lit, bnh hn m gan.

L-AG cn dng lm nguyn liu khi u cho vic tng hp mt s ho cht quan
trng: N-acetylglutamat l cht hot ng b mt, vi sinh vt c th phn gii c,
t n da, c dng rng ri trong cng nghip m phm, x phng v du gi u.
Axit oxopyrolidicarboxylic, mt dn xut khc ca L- AG c dng lm cht gi
m trong m phm. Acetylglutamat c dng trong x l nhim nc bin do
du ho v du thc vt gy nn. L-AG phn b rng ri trong t nhin di dng
hp cht v dng t do, c trong thnh phn cu to ca protein ng thc vt.
Trong m L-AG to thnh t NH3 v axit - xetoglutaric. Trong 6 sinh vt, c bit
l vi sinh vt, L-AG c tng hp theo con ng ln men t nhiu ngun cacbon.
Vai tr ca m chnh Khi trung ho axit glutamic chuyn thnh glutamat natri (m
chnh), kt tinh c v ngt du trong nc, gn ging vi v ca tht. Glutamat natri
c ngha ln i vi i sng con ngi, n c s dng cc nc Trung Quc,
Nht Bn, Vit Nam... Cc nc chu u ch yu dng m chnh thay mt phn
tht cho vo cc hn hp thc phm, xp, ru, bia v cc sn phm khc. M chnh
l cht iu v trong ch bin thc phm, lm gia v cho cc mn n, cho, m n
lin, tht nhn to, cc loi tht c ng hp v. v... nh sn phm hp dn hn v
L- AG c a vo c th, lm tng kh nng lao ng tr c v chn tay ca con
ngi. Cc nghin cu khoa hc ch ra rng, glutamate ng vai tr quan trng
trong c ch chuyn ho cht b dng trong c th con ngi. Trn thc t, c th
ca mi ngi cha khong 2 kilogram glutamate c tm thy trong cc c bp,
no, thn, gan v cc c quan khc. Lng glutamate c trong c th ngi dng
t do v lin kt l khong 2000 g. Lng glutamate t do c trong c th ngi l
10 g, trong :
+ C bp : 6.0 g.
+ No : 2.3 g.
+ Gan : 0.7 g.
+ Thn : 0.7 g.
+ Mu : 0.04 g.
Cc nghin cu khoa hc cng cho thy rng glutamate t nhin c trong thc
phm v glutamate c ngun gc t m chnh u ging nhau. Chng c h thng
rut hp th v tiu ho nh nhau. Mt khi c tiu ho, c th chng ta khng
phn bit c u l glutamate t thc phm hay t m chnh. Thc t nghin cu
cho thy rng glutamate t thc phm hay t m chnh u quan trng i vi chc
nng ca h tiu ho.

III. Cc phng php sn xut m chnh

M chnh d c sn xut bng phng php no cng thng tun theo mt s
tiu chun sau:
- Tinh th MSG cha khng t hn 99% MSG tinh khit.
- m (tr nc kt tinh) khng c cao hn 0,5%.
- Thnh phn NaCl khng c qu 0,5%.
- Cc tp cht cn li khng cha Asen ,kim loi v hp cht Canxi.
C nhiu phng php sn xut m chnh khc nhau, t cc ngun nguyn liu
khc nhau. Hin nay, trn th gii c 4 phng php c bn:
1. Phng php tng hp ho hc
Phng php ny ng dng cc phn ng tng hp ho hc tng hp nn axit
glutamic v cc aminoaxit khc t cc kh thi ca cng nghip du ho hay cc
ngnh khc. V d: Nht nm 1932 tng hp c 300 tn axit glutamic, prolin
v.v... t cracking du ho, t furfurol tng hp ra prolin, lizin.
- u im: Phng php ny c th s dng ngun nguyn liu khng phi thc
phm sn xut ra v tn dng c cc ph liu ca cng nghip du ho.
- Nhc im: Ch thc hin c nhng nc c cng nghip du ho pht trin
v yu cu k thut cao. Mt khc sn xut bng con ng ny to ra mt hn hp
khng quay cc D, L-axit glutamic, vic tch L-axit glutamic ra li kh khn nn
lm tng gi thnh sn phm. Do nhc im nh vy nn phng php ny t c
ng dng cc nc.
2. Phng php thu phn protit
Phng php ny s dng cc tc nhn xc tc l cc ho cht hoc fermen
thu phn mt ngun nguyn liu protit no (kh u, kh lc) ra mt hn hp
cc aminoaxit, t y tch cc axit glutamic ra v sn xut m chnh. Qu trnh ny
c th tm tt nh sau: gluten ca bt m c thy phn bng axit HCl gii
phng ra tt c cc axit amin 1500 C. Sau cc cht cn b s c lc, dch lc
c c c v gi nhit thp lm gim ha tan ca cht tan, t cc
ht tinh th kt tinh ca hydroclorat glutamic Natri HOOC- CH 2- CH2- CH-COOH
qu bo ha s dn dn c
|

CH3

to thnh.
Nhng ht tinh th ny s c lc tch ring v sau c ha tan trong
nc. Dung dch ny s c trung ha bng Na2CO3 cho ti pH = 3,2 (pH ng

in), pH ny tinh th axit glutamic s kt tinh ra khi dung dch v c tch

ring bng phng php ly tm. Sau pha long v kt tinh ln 2 vi dung dch
Na2CO3 pH = 5,7 7,0. Than hot tnh v Na2CO3 c thm vo kh mu
v kt ta cc tp cht. Tp cht s c lc, dch lc c c c bng phng
php bay hi chn khng thu c dch c c MSG, dch c c c tch nc
bng phng php ly tm, sn phm thu c c sy kh to nn tinh th cui
cng l MSG tinh khit. Hiu sut thu hi MSG thay i trong khong 15% 25%
khi s dng bt m. i vi u nnh th hiu sut thu hi MSG thp hn rt nhiu
ch khong 4% 7%. Hin nay nc ta v nhiu nc trn th gii ch yu vn s
dng phng php ny.
- u im : D khng ch quy trnh sn xut v p dng c vo cc c s th
cng, bn c gii, c gii d dng.
- Nhc im: + Cn s dng nguyn liu giu protit him v t.
+ Cn nhiu ho cht v cc thit b chng n mn.
+ Hiu sut thp, a n gi thnh cao.
3. Phng php ln men
Phng php ny li dng mt s vi sinh vt c kh nng sinh tng hp ra cc axit
amin t cc ngun gluxit v m v c. Phng php ny ang c nhiu trin vng
pht trin khp cc nc, n to ra c nhiu loi aminoaxit nh: axit glutamic,
lizin, valin, alanin, phenylalanin, tryptophan, methionin ...
Phng php ln men c ngun gc t Nht Bn, nm 1956 khi m Shukuo v
Kinoshita s dng chng Micrococcus glutamicus sn xut glutamat t mi trng
c cha glucoza v amoniac. Sau mt s loi vi sinh vt khc cng c s dng
nh Brevi bacterium v Microbacterium.
Tt c cc loi vi sinh vt ny u c mt s c im sau:
+ Hnh dng t bo t hnh cu n hnh que ngn
+ Vi khun Gram (+)
+ H hp hiu kh
+ Khng to bo t
+ Khng chuyn ng c, khng c tin mao
+ Biotin l yu t cn thit cho sinh trng v pht trin
+ Tch t mt lng ln glutamic t hydrat cacbon v NH4 + trong mi trng c sc
khng kh.
Khi s dng Micrococcus glutamicus c nhiu cng thc thit lp mi trng nui

cy khc nhau, di y chng ti a ra 2 cng thc lm v d :

Tanaka (g/l) Ajinomoto (g/l)
Glucoza

100

Ur

KH2PO4

100
8

MgSO4.7H2O

0,1
0,25

0,04

Dch thy phn u nnh

Cao ng

2,5

0,5

Nit amin
Biotin

25

Fe v Mn

0,5

0,2

Thi gian ln men

35 h

Hiu sut thu hi

50

40 h
44,8

Nhit ln men gi 28oC v duy tr pH = 8,0 bng cch thng xuyn b sung
ur. iu kin hiu kh l rt quan trng bi v nu khng c sc kh th sn phm
to thnh khng phi l axit glutamic m l lactat. Khi s dng nguyn liu ln men
l r ng th cn phi b sung cc cht khng biotin kim sot s sinh trng
ca vi sinh vt. Phng php ny c nhiu u im nn ang c nghin cu v
ng dng nc ta v cc nc trn th gii.
- u im chnh: + Khng s dng nguyn liu protit.
+ Khng cn s dng nhiu ho cht v thit b chu n mn.
+ Hiu sut cao, gi thnh h.
+ To ra axit glutamic dng L, c hot tnh sinh hc cao.
4. Phng php kt hp
y l phng php kt hp gia tng hp ho hc v vi sinh vt hc. Phng
php vi sinh vt tng hp nn axit amin t cc ngun m v c v gluxit mt nhiu
thi gian, do ngi ta li dng cc phn ng tng hp to ra nhng cht c cu
to gn ging axit amin, t y li dng vi sinh vt tip tc to ra axit amin. Phng
php ny tuy nhanh nhng yu cu k thut cao, ch p dng nghin cu ch t p
dng vo cng nghip sn xut.
IV. C ch to axit glutamic ca chng Micrococcus glutamicus.
S sinh tng hp axit glutamic v cc aminoaxit khc tmi trng nui cy vi
khun to ch yu axit glutamic:

4.1 T ngun cacbon l sacharide theo chu k Embden- Mayerhaf:

Lc ny t A.pyruvic to ra Acetyl-CoA

Tt c cc loi saccharide u cho ta sn phm phn ng l axit - cetoglutaric. T

trong mi trng c ngun N th xy ra phn ng bi cc men to ra axit
glutamic.
Ngoi ra mt saxitamin khc cng c to thnh tcc sn phm trung gian ca
qu trnh phn hu ng v l sn phm phca dung dch ln men.
4.2. Cc sn phm ca qu trnh ln men L-AG
4.2.1. Sn phm chnh
Phng trnh tng qut ca qu trnh to L-AG tglucoza hay axetat v NH3 c
biu din nhsau :
C6H12O16+ NH3+ 1,5 O2 L-AG + CO2+ 3 H2O

3CH3COOH + NH3+ 1,5 O2 L-AG + CO2+ 3 H2O

Theo phng trnh ny th sn phm chnh l L-AG v CO2. Trong chc L-AG
l c quan tm v ngha kinh t ln lao ca n. y theo l thuyt, hiu sut
chuyn ho (HSCH) glucose hay axetat thnh L-AG u l 81,66%. Thc t nghin
cu v sn xut cha bao gi t c gi tr ny phn v c cht cn d li trong
mi trng, phn v phi dng cho tng sinh khi v to cc sn phm khng mong
mun ngoi L-AG. Theo Kinoshita v cng s, HSCH c th chp nhn c khi
a phng php ln men L-AG t glucose vo sn xut cng nghip l 30%. Ngy
nay, tu theo iu kin sn xut v phng tin qu trnh ln men ngi ta nng
c hiu sut chuyn ha ng thnh L-AG l 45 50 % trong sn xut v 55
57% ging t nhin hay 61 62% t ging t bin trong nghin cu phng th
nghim. Nhvy so vi hiu sut chuyn ha l thuyt, hiu sut thc t phng th
nghim mi t c 76% t glucose v 70% t benzoat. Ngi ta ang tm mi
bin php rt ngn khong cch gia hiu sut chuyn ha l thuyt v hiu sut
chuyn ha thc t.
4.2.2 Sn phm ph
a. Axit lactic
Trong iu kin ti u, L-AG sinh ra l ch yu. Nu chch khi iu kin ny th
Corynebacterium glutamicum s to axit lactic thay v to L-AG. C hai l do cbn
dn ti tnh trng ny, hoc l qu d tha biotin hoc l qu t oxy ho tan. i khi
s thay i nhit t ngt t 30 n 37 C cng dn ti vic bin qu trnh ln
men L-AG thnh qu trnh ln men axit lactic nh xy ra vi B. divaricatum.
b.Axit sucxinic
Tng t nh axit lactic, axit sucxinic c sinh ra vi s lng ln khi cung cp
tha biotin hoc cung cp t oxy ho tan, c bit nhiu khi va tha biotin va thiu
oxy ho tan. Ngun gc ca s tch t axit sucxinic l do axit fumaric b kh bi
coenzim NADH l cht cho hydro. V vy cn phi cung cp oxy ho tan v gii
hn nng biotin phn ng va ni t c c hi din ra.
c.Axit - xetoglutaric
Mi qu trnh sinh tng hp u c phn ng to L-AG t -xetoglutaric nh xc
tc ca hai h thng enzim transaminase v L-AG dehydrogenase. Phn ng ny
thc hin c hon ton khi mi trng c d NH4+ v pH t trung tnh n kim
yu. Nu mi trng thiu NH4+ v pH phm vi axit yu th phn ng trn khng
thc hin c. Kt qu l -xetoglutaric b tch t ngy mt nhiu trong mi trng

thay v L-AG. Ngi ta thy -xetoglutaric c hnh thnh ch yu t axit citric v

trong t bo di iu kin hn ch nng biotin v NH4+ ri mi thi ra ngoi
mi trng. Trong trng hp ln men L-AG t n-alkan nh chng
Corynebacterium hydrocacboclastus S10B1 -XG sinh ra nhiu nh hn ch nng
B1ca mi trng. Vic ny gy nn thiu ht TPP cn cho hot ng ca enzim,
xitrat-decacboxylaza, hn ch isocitrat i vo chu trnh glyoxylat, thc y isocitrat
i vo chu trnh TCA v sn sinh ra -XG.
d.Glutamin v sn phm khc
Ngi ta nhn thy trong tt c cc qu trnh ln men sn xut L-AG u c
glutamin (GM), L-acetylglutamin (L-AGM), alanin v aspartic trong dch men vi
s lng khc nhau tu thuc vo loi ging v iu kin nui dng chng hoc
thay i cu to mi trng u c th chuyn qu trnh sn xut L-AG thnh qu
trnh sn xut glutamin nh cng mt ging. Trong iu kin bnh thng glutamin
c tng hp nn nh enzim glutamin-systhetaza. Enzim ny hot ng tt pH
axit v khng b km hm bi (NH4)2SO4 nng cao. Mt loi enzim khc cng
xc tc qu trnh to glutamin, l glutaminase. Nhng enzim ny hot ng tt
pH kim v nng thp ca (NH4)2SO4, nhng b km hm bi (NH4)2SO4
nng cao. Nakanishi v cng s cho bit chng Corynebacterium glutamicum
KY9609 sinh lng ln GM v L-AGM bn cnh L-AG di iu kin mi trng
c nng biotin, NH4Cl v ion Zn2+thch hp. y c GM ln L-AGM u tng
ln v L-AG gim xung khi mi trng c pH axit yu sau giai on sinh trng.
Nu dng (NH4)2SO4vi nng 4% lm ngun cung cp NH 3 ngoi ur th lng
glutamin sinh ra bng lng L-AG. Nu thay (NH 4)2SO4 bng NH4Cl vi cng nng
th lng GM sinh ra p o lng L-AG v qu trnh ln men L-AG
hon ton chuyn thnh qu trnh ln men GM. Hiu sut ln men GM cng tng
cao khi mi trng c thm ion Zn2+ hoc Zn2+ cng vi Ni2+ v Cr6+ vi nng
thch hp v c th t ti 40g t 133g glucose (HSCH l 30%). Yoshihaza, v
cng s pht hin ra rng nu thm vo mi trng mt trong bn cht MG, DMG,
TMG v HETMA v c bit thm c cht thi stephen, mt sn phm ph cc
cng on sn xut ng t c ci ng, th hiu sut ln men cc axit amin v 5inosinic tng ln rt nhiu v c th sn xut cL-AG ln GM nh cng mt chng
C. acetoacidophilumATCC 13870 [224]. Gn y, Tsuchida v cng s to c
ging B. flavumAJ 12418 hoc C.acetoacidophilumAJ 12419 bn vng vi cc
dipeptit (tyrosin-glutamic hoc alanin-glutamic) c kh nng tng hp mt lng

ln GM khi c mt ca mt trong bn dipeptit ni trn. Suzuki v cng s tm

thy s c mt ca trehalose v glucose trong dch men ca chng Athrobacter
parafineus sinh trng trong mi trng n-parafin C13-C21 khi c thm PG vo
giai on u ca qu trnh sinh trng. y tng lng ng tng lin tc theo
thi gian ln men nhng ng kh glucose tng rt t. Nh vy s cn li l ng
khng c tnh kh, ng trehalose. Suzuki v cng s cho bit vic b sung
Cu2+vo mi trng vi lng 1,26 mg/l v b sung PG vo giai on u sinh
trng ca Athrobacter parafineusKY 4303 trn mi trng parafin C13-C21 lm
tng tch lu L-AG theo thi gian. Gn y, Yoshii, H. v cng s thng bo rng
c th iu tit s tch lu trehalose lm tng sn lng L-AG trong ln men.
Okazaki v cng s khng nh c ti 35 45% axit oleic a vo mi trng
c B.thiogenitalisNo 653, mt th t bin vn i hi axit oleic cho sinh
trng v tch lu L-AG, li dng to thnh hp cht ng - lipit. Nakao v
cng s cho bit nu thm PG hoc cephaloridin vo cc t bo C.alkanolyticum
No.314 sinh trng trn n-parafin th s gy nn s tch t ng thi ba sn phm
photpholipit, dn xut N-acetyl-hexoamin, (mt hp cht trung gian to nn thnh t
bo) v L-AG trong gia L-AG v photpholipit ngoi bo c mi tng quan
cht ch: L-AG t l thun vi photpholipit ngoi bo. Kikuchi v cng s nghin
cu tip vn ny v xc nhn cc axit bo C14-C18cng c tch t ngoi bo
cng vi photpholipit; thnh phn cu to ca photpholipit ngoi bo v ni bo u
ging nhau. Vic tch lu N-acetyl hexozamin ngoi bo c lin quan vi bn cht
ca cht khng sinh a vo t bo. Tt c cc khng sinh c ch tng hp mng t
bo u thc y vic bi tit cc cht cu to nn t bo vo trong mi trng.
Trong s7 khng sinh em th D-cycloserin c tc dng mnh nht. N-acetyl
hexozamin l nhng sn phm trung gian cu to nn thnh t bo. Ngi ta bit
hp

cht

ny

bao

gm

thnh

phn

axit

N-acetylmuramic,

Nacetylhexozaminuronic v N-acetylglucozamin .
4.2.3.S chch hng to sn phm chnh
Trong iu kin bnh thng sn phm chnh ca mi qu trnh sinh tng hp LAG l L-AG v CO2 nh ni trn. Nhng khi thay i iu kin ln men th
sn phm chnh khng phi l L-AG m l cc sn phm khc. Takamura v cng s
cho bit thay i lng cao nm men v cao ng gi cho nng photphat v c
trong mi trng phm vi trn di 0,0129% slm cho vi khun Acetobacter
aerogenes khng sn sinh L-AG m sn sinh valin vi s lng ln. Asaki v cng

s khng nh c th hn ch nng photphat v c bng cch thm 6mercaptopurin cho qu trnh ln men LAG vi khun trn chuyn thnh qu
trnh ln men valin. Kinoshita S. v cng s gy t bin cc ging sinh L-AG
to ra ging mi c th sinh lysin mi trng giu biotin. Nara v cng s
nghin cu tc dng ca PG ln vic chuyn sn phm chnh cc chng sinh
homoserin, lizin hay valin v thy rng nu thm 4 v/ml PG vo gi th7 9 sau
khi bt u ln men nh chng sinh homoserin M. Glutamicus 534 - Co147 th qu
trnh ln men homoserin s chuyn thnh qu trnh ln men L-AG. Hin tng
tng t cng xy ra i vi ging sinh lysin v valin. Cc ging ny tch lu L-AG
thay v lysin hay valin khi c thm PG vi lng v thi im nh ni trn.
Sau ny, Shiratsuchi v cng s xut mt quy trnh ln men mi bin qu trnh
ln men lysin thnh qu trnh ln men c lysin ln L-AG. Nguyn tc ca phng
php ny rt n gin: cc tc gi sdng ging sinh lysin ln men trong mi trng
giu biotin kt hp b sung cht hot ng b mt hoc PG (tng t khi ln men
bng ging sinh L-AG) v iu bt ng xy ra: c lysin v L-AG u c tch t
vi s lng ln ti mc c th p dng tt trn quy m cng nghip. Phng php
mi mang li hiu qu trn 2 phng din. Mt l tng lng lysin v LAG sinh
ra u nhiu gp 1,3 1,45 ln phng php c, hai l pH mi trng t b thay i
nh skt hp gia lysin (mang tnh kim) v L-AG (mang tnh axit).
V. K thut sn xut axit glutamic v bt ngt
5.1. Nguyn liu
ln men sn xut axit glutamic, ngi ta dng nguyn liu ch yu l dch c
ng, hoc r ng, hoc cc ngun nguyn liu tinh bt qua giai on ng
ha. Khoai m l nguyn liu tinh bt c s dng nhiu nht hin nay. Ngoi ra
cn c cc ngun dinh dng b sung nh mui amn, photphat, sulfat, biotin,
vitamin B
Trong thc t sn xut, ngi ta dng r ng lm mi trng ln men thay cho cao
bp. R ng thng pha long n 13 14% v thanh trng trc khi ln men.
Nu l nguyn liu cha tinh bt, th tinh bt phi c thy phn (qu trnh dch
ha v ung ha) nh enzym a -b- amylaza ri sau mi b sung thm dinh
dng vo mi trng ln men.
5.2. Chng vi sinh
Tham gia vo qu trnh ln men sn xut axit glutamic, chng vi sinh thng s
dng l: Corynebacterium Glutanicum, Brevibacterium Lactofermentus, Micrococus

Glutamicus; nhng ch yu nht vn l chng Corynebacterium Glutamicum (loi vi

khun ny c nh vi sinh vt Nht Bn Kinosita pht hin t 1956, c kh nng
ln men t tinh bt, ng, khoai, khoai m to ra axit glutamic).
Ging vi khun thun khit ny c ly t ng thch nghing ti cc c s gi
ging, sau c cy truyn, nhn sinh khi trong mi trng lng (nh ni
phn trn). Khi lng sinh khi uc nhn ln n yu cu ph hp cho quy trnh
sn xut i tr. Trc khi nhn, cy, mi trng lng phi c thanh trng bng
phng php Pasteur.
Chng vi khun ging phi c kh nng to ra nhiu axit glutamic, tc sinh
trng pht trin nhanh, c tnh n nh cao trong thi gian di, chu c nng
axit cao, mi trng nui cy n gin, d p dng trong thc t sn xut.
Axit glutamic sn xut bng phng php ln men vi khun, vi nguyn liu l
ng. Qu trnh ny c xc tc nh h enzym c sn trong vi khun, chuyn ha
qua nhiu giai on trung gian vi nhiu phn ng khc nhau to ra nhiu sn phm
ph, v cui cng l sn phm axit glutamic. Thc cht ca qu trnh ny l ng
c chuyn ha (qu trnh ng phn theo Enbden Meyerhoff), ri sau thng
qua chu trnh Krebs ca qu trnh h hp hiu kh ca vi khun, sn phm axit
glutamic c hnh thnh. S hnh thnh axit glutamic ph thuc vo s tch t axit
a - xtoglutaric trong t bo vi khun, s c mt ca NH 3 v enzym xc tc l
glutamat dehydrogenaza.
Phng php ln men vi khun l phng php c s dng rng ri hin nay trn
th gii sn xut axit glutamic v bt ngt. Hng nm, sn lng bt ngt c th
gii sn xut theo phng php ny khong 25 30 vn tn. Vit Nam cng c
nhiu nh my sn xut bt ngt bng phng php ln men nh VeDan, Ajino
Moto, Vit Tr, Thin Hng
sn xut bt ngt t axit glutamic bng phng php ln men, quy trnh cng
ngh c trin khai theo cc giai on sau:

Chun b dch ln men: Mi trng ln men c chun b sn t cc nguyn

liu ng hoc tinh bt (nh nu phn trn) c thanh trng k trc khi cy
vi khun ln men glutamic vo.

Giai on ln men: dung dch nhn sinh khi vi khun, dung dch ln men c

chuyn vo cc dng c, thit b ln men, sau corynebacterium glutamicum vo, cho

ln men trong iu kin thong kh, gi nhit 32 370C trong thi gian 38 40
gi. Kt thc qu trnh ln men, lng acid glutamic c th t 50 60g/ lt.

Trong thi gian ln men, pH s chuyn dn sang acid do s hnh thnh axit glutamic
do ngi ta thng b sung thm dinh dng vo mi trng ngun amoni
(NH4Cl, (NH4)2SO4, ur) gi n nh pH cho vi khun hot ng tt.
Khng c iu kin ln men l ym kh v sn phm to ra s l axit lactic.
to thong kh, trong cc thit b ln men b tr b phn khuy trn dch vi tc V
= 450 vng/ pht.

Tinh sch acid glutamic:

Kt thc qu trnh ln men, acid glutamic c to thnh cng vi mt s tp cht

khc, do cn phi tinh ch cc tp cht ny ra khi dung dch cha acid glutamic.
Phng php thng dng l nha trao i rezin. Nha trao i rezin c hai loi:
rezin dng tnh (mang tnh axit) v rezin m tnh (mang tnh kim).
Dch ln men c cha axit glutamic v tp cht cho chy qua ct nha (c cha
rezin) t di ln vi tc 150 180 lt/ pht, thi gian chy qua ct l 150 180
pht. Song song, ngi ta cho dng nc chy qua ct cng chiu vi dung dch ln
men ra cc vi khun bm vo b mt rezin. Gi nhit trong ct trao i ion l
600 650C. Sau khi kt thc qu trnh trao i ion, dng NaOH 4 5% tch axit
glutamic ra khi ct (tc chy NaOH l 5 6m/ gi, lu lng 100lt/ pht).
Ngi ta c th s dng than hot tnh kh mu. Axit glutamic c thu bng
cch iu chnh pH = 3,2 ri c c dung dch v gim nhit xung 40 150C s
thu c tinh th acid glutamic vi lng 77 88% hoc cao hn.

S to thnh bt ngt:

Bt ngt l mui natri ca axit glutamic, gi l glutamat natri. Dng NaOH 40%
50% trung ha dung dch axit glutamic n pH = 6,8, sau em lc, c c, v
kt tinh bng phng php sy chn khng nhit thp s thu c tinh th bt
ngt mu trng. tinh khit ca bt ngt c th t 99 99,6% mononatri
glutamat.
VI. c tnh v liu lng s dng:
Trong mt hi tho ti H Ni do Vin Dinh dng t chc, cc chuyn gia thuc
y ban K thut Glutamate quc t cho bit: bt ngt c th b bin i thnh cht
gy t bin gen khi thc hin th nghim trong hai gi ng h nhit 300 0 C.
Nhng chng ta nn nh rng: khng ch bt ngt b bin i m nhit ny, cc
thnh phn thc phm t nhin nh protein cng u b bin i thnh cht gy t
bin gen, cc nguyn liu thc phm b nu nhit cao cng b chy thnh than.
Tuy nhin, nhit si ca nc v thc n c ch bin hng ngy, natri v

Glutamat - hai thnh phn chnh ca bt ngt vn l nhng cht n nh.

Hng trm cuc th nghim ton din trn ng vt v c c th ngi c thc
hin bi cc vin nghin cu hng u trn th gii trong mt thi gian di a ra
kt lun: Bt ngt m bo an ton cho mc ch s dng i vi mi la tui, c
th nh sau:
- y Ban Chuyn gia v Ph gia Thc phm (JECFA) ca T chc Y t Th gii
(WHO) v T chc Lng nng Quc t (FAO) cng b trong cun Hng dn s
dng an ton cc cht ph gia thc phm (Guide to the Safe Use of Food Additives)
xut bn nm 1979 th bt ngt thuc danh sch A1 lit k cc ph gia c y
Ban JECFA hon ton thng qua cho php dng an ton (fully cleared) vi liu dng
hng ngy chp nhn c l 0-120mg/kg. C ngha vi th trng ngi bnh
thng 50kg th mi ngy c th s dng khong 6g bt ngt.
-Nm 1991 u ban Khoa hc v thc phm ca Chu u ti khng nh bt ngt an
ton v c th dng hng ngy.
-Nm 1992 bo co t hi ng Khoa hc thuc hip hi Y t M khng nh di
bt k hnh thc no cng khng chng minh c n l mi nguy hi ng k i
vi sc kho.
-Nm 1995 bo co c FDA u quyn ca hip hi Thc nghim sinh hc lin
bang (FASEB) xc nhn cc nh gi v an ton ca cc u ban nu, nhng cho
rng mt t l cha r dn s c th d ng vi bt ngt v pht trin thnh mt hi
chng phc tp khi tiu th hn 3g bt ngt thun.
-Nm 2002 mt bo co pht hin nhng con chut c nui theo ch dinh
dng b sung t 10 20% bt ngt tinh khit b thoi ho vng mc, c th do s
dn t ca mui glutamate trong pha l dch. Tuy nhin, mt lng glutamat cao
hn rt nhiu so vi lng bt ngt s dng lm gia v thc phm.
- cc nc trn th gii:
+ Ti Hoa k: bt ngt c cc c quan qun l thuc v thc phm Hoa K a
vo danh sch cc gia v thc phm c php s dng. Bt ngt c coi l an ton
cho mc ch s dng v c xem l mt thnh phn thc phm ph bin nh
mui, tiu, gim, bt n.... v khng quy nh liu dng hng ngy (Theo ti liu
Code Federal Regulation Part 182 - 1994. FDA - US )
+ Ti Php: Bt ngt c coi l an ton v cng khng quy nh liu dng hng
ngy. (Theo ti liu Rglementation des produits qualit - Repression des fraudes
1991 et modifi 6/1993).

+ Ti Thi Lan: liu dng hng ngy ty theo yu cu (As required).

+ Ti cc nc Chu khc: Malaysia, Philippin, Nht, Trung Quc, i Loan, Hn
Quc u coi bt ngt l an ton v khng quy nh liu dng hng ngy.
+ Cng ng Chu u ( EC) t m s bt ngt (MSG) l ph gia thc phm s
E621 v cho php dng khng c khuyn co s lng hng ngy.
+ Vit Nam
- B Y T: bt ngt c xem l mt gia v thc phm v c a vo danh
mc cc ph gia thc phm c php s dng trong ch bin thc phm theo quyt
nh s 3742/2001 Q- BYT ngy 31/8/2001 ca B Y T.
- B khoa hc cng ngh v mi trng: bt ngt c php s dng nh mt ph
gia thc phm trong ch bin thc n gia nh, ti cc nh hng cng nh trong
cng nghip ch bin thc phm (Bo co ngy 20/4/1995 ca Tng Cc tiu chun
o lng cht lng - B khoa hc Cng ngh v Mi Trng).
VII. Bt ngt c lin quan g n Hi chng qun n Trung Hoa khng?
Hi chng qun n Trung Hoa (Chinese Restaurant SyndromesI) l cm t dng
ch triu chng kh chu nht thi do b d ng khi n ti cc qun n Trung Hoa.
H cho rng trong cc thc n c nhiu bt ngt gy nn hin tng nng mt, hoa
mt, kh chu.... Nm 1968, bc s Ho Man Kwok thuc t chc National Biomedial
Research Foundation vit mt bi bo cho chuyn san y t New England of
Medecine nh sau: Ti b mt hi chng l mi khi i n trong mt nh hng Tu,
c bit l nh hng bn thc n Hoa. Hi chng thng bt u din ra khong 15
20 pht sau khi n xong mn u tin, ko di khong hai gi ng h m khng
c cc hu qu li. Nhng triu chng ni r nht l b t cng pha sau gy, dn
dn lan to ra hai cnh tay v lng, b u oi v tim p nhanh Li thut ny bt
u gy hoang mang v sc kho trn khp th gii v bt ngt v t Hi chng
qun n Trung Hoa ra i t y. Tuy nhin, ng v cc cng s nghin cu v
pht hin ra rng cc triu chng trn c th gy ra bi mt s thnh phn c trong
nc tng. Nhng triu chng ny li khng xy ra khi s dng loi nc tng
ny nh. Mt vi nhn nh rng triu chng gy ra c th do ru c s dng
rng ri trong mn n cc nh hng Trung Quc, v cc triu chng ny tng t
nh cc triu chng do nh hng ca ru. Nhn nh khc li cho rng triu
chng ny gy ra bi bt ngt c s dng lm gia v ch bin trong cc mn n
nh hng Trung Quc. V mt kh nng khc l hm lng mui cao trong cc mn
n Trung Quc c th lm cao natri mu nht thi, gy nn hin tng gim kali

trong ni bo, to ra cc triu chng t c, hi hp v c cm gic yu

Cn theo tin s Kenny ca trng i Hc Y Khoa George Washington lm th
nghim i chiu lm sng v vn ny, kt qu ca th nghim chng minh:
bt ngt khng h lin quan trc tip ti d ng. Nghin cu ca ng cn a ra kt
lun, n xong ung nc c chua v c ph en, cng c th gy nn kh chu sau
khi n mt s thc phm hoc mt s thnh phm ca thc phm no , nhng
thc phm ny thng l ht tiu, scla, cc loi s hn v nhiu loi thnh phn
thc phm khc.
Cho n nay, da trn cc nghin cu khoa hc ng tin cy, y ban hn hp v ph
gia thc phm (JECFA) ca T Y T Th gii (WHO) v t chc Lng Nng
(FAO) nm 1987, chnh thc tuyn b: Cc nghin cu khng th chng minh
c rng MSG (bt ngt) l tc nhn gy ra cc triu chng ca Hi chng qun
n Trung Hoa.
Trong thc t i sng hng ngy ta cng c th gp mt s ngi khi n c bin,
tm, cua hay gh, tht g, tht b ......cng sinh ra d ng mn nga hoc kh chu.
cng l do c a mn cm ca tng ngi d ng vi mt loi thc phm khc
nhau. Nn khng th ni bt ngt l tc nhn gy ra ton b Hi chng qun n
Trung Hoa i vi tt c mi ngi khi cha chng c chc chn. Qua cc quy
nh quc t v quc gia c s dng bt ngt (MSG), chng ta nhn thy khng c
quc gia no cho bt ngt l cht c v cng khng c quc gia no cm s dng
bt ngt trong ch bin thc phm. Tuy nhin, cn xc nh r bt ngt ch l mt
ph gia thc phm, iu v an ton cn thit (tng t nh dm, tiu, mui n...),
bn thn bt ngt khng phi l mt cht dinh dng c th thay th cho tht, c,
trng, sa....
VIII. Lu khi ch bin:
Khng nu nhit caoKhi cho thm bt ngt vo thc n c nu nhit
cao s xy ra thay i ha hc, khin bt ngt tr nn c hi cho sc khe. 70 90oC l nhit thch hp nht ha tan bt ngt. V vy nn gia gim bt ngt
khi thc n chn v bc khi bp.
Khng cho trc tip khi thc phm nhit thpBt ngt ha tan km nhit
thp. Nu bn mun s dng bt ngt tng v ngon cho mn ngui th nn ha tan
bt ngt trong nc m ri mi trn vo thc n ngui.
Khng cho vo cc thc phm ngtTuyt i khng nn thm bt ngt vo thc
phm c v ngt t nhin (c chua, tm.) v s lm mt hng v, ngt ca mn

n v gy v kh n.
Cho qu nhiuCng nh cc gia v khc, lng bt ngt d tha s khin mn n
b mt v v cn c hi cho sc khe. Khng cho vo cc mn chin rn rau c v
thc phm c mu vngV nh vi mn trng, trong trng c nhiu bt v khi kt
hp vi mui natri clorua ri un nng s to ra th bt ngt tinh khit, gip trng
c hng v thm ngon. V th cho bt ngt vo trng l tha v cn khng tt cho
sc khe.
Gn y trn th trng cn xut hin nhiu loi bt nm c kh nng to ngt gp
nhiu ln so vi m chnh. Theo cc nh chuyn mn, cc loi bt nm trn cng
chng c gi tr dinh dng g, nhng li cho o gic kh cao, lm tng hng v ln
hng chc ln nh vo cc nucleotid v mui natri. Vic s dng chng s khng c
li cho sc khe. Ngoi ra, hin nay xut hin cc loi m chnh gi nhi nhn hiu
ca cc hng ni ting v pha thm nhng cht c hnh dng tng t nh hn the,
phn. Cc cht ny l tc nhn gy tn hi cho d dy, gan, lm cho con ngi km
n, kh chu ton thn v c th gy ung th bng quang.

TI LIU THAM KHO:

Cng ngh sn xut m chnh v cc sn phm ln men c truyn Trng i hc
Bch khoa H Ni GS.TS Nguyn Th Hin (Ch bin).
Ph gia thc phm m Sao Mai (Ch bin), Nguyn Th Hong Yn, Bi ng
Khu.
http://www.amthucchay.org/2013/05/tim-hieu-ve-bot-ngot-bot-nem-va-suckhoe.html.
http://doc.edu.vn/tai-lieu/de-tai-san-xuat-glutamate-9966/