Nghiên cứu điều chế magie lactat bằng phương pháp sinh tổng hợp

B Y T
I HI



NGUYN TH THU TRANG

U CH MAGIE
LACTAT B
TNG HP


N TT NGHI



I - 2014
B Y T
I HI



NGUYN TH THU TRANG

U CH MAGIE
LACTAT B
TNG HP


N TT NGHI

ng dn:
1. TS. Đàm Thanh Xuân
2. DS. Nguyễn Minh Ngọc
c hin:
1. Bộ môn Công nghiệp dược
2. Trường Đại học Dược Hà Nội


I - 2014
LI C

Vi s i li
c          DS. Nguyn Minh Ngc,
nhng  trc ting d em trong su


Em i li c
ch k thu 
 u kin thun li cho em trong sun.

Em rt mong nha th 
thiu khoa hc c





Nguyn Th Thu Trang

MC LC

LI C
MC LC
DANH M VIT TT
DANH MNG
DANH M  TH
T V
NG QUAN
1.1. Magie lactat
1.1.1. 
1.1.2. 
1.1.3. Mt s sn ph 
1.2. n xut magie lactat
1.2.1. ng h
1.2.2. 
1.3. Vi khun sinh axit lactic
1.3.1. 
1.3.2. Chi Lactobacillus
1.3.3. Lactobacillus acidophilus
1.3.4. 
1.4. 
U
t liu, thit b
t li
2.1.2. Thit b s d

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2.2. Nu
2.2.1.  
2.2.2. Kht s  
u

ng h
 d
2.3.4. lm
p mdihydrat
 

u su 
2.3.8
C NGHIM, KT QU N.
3.1.  o magie lactat 
3.1.1. La ch
3

3.1.2. La chn ph
3.1.3. n phm m
3.2. Kht s  
3.2.1. Khng c
3.2.2. Khng ca n 
KT LU XUT
U THAM KHO
PH LC
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DANH M VIT TT
ATCC
American Type Culture Collection
C%
N ph

n Vit Nam
E. coli
Escherichia coli
FAO
Food and Agriculture Organization
Glc
Glucose
H
Hiu sut
L. acidophilus
Lactobacillus acidophilus
L. brevis
Lactobacillus brevis
L. buchneri
Lactobacillus buchneri
L. bulgaricus

Lactobacillus bulgaricus
L. casei
Lactobacillus casei
L. delbrueckii
Lactobacillus delbrueckii
L. fermentum
Lactobacillus fermentum
L. leichmenii
Lactobacillus leichmenii
L. plantarum
Lactobacillus plantarum
S. lactis
Sacharomyces lactis
M
Kh

 lch chun
R
f
H s di chuyn
x
 

V
WHO
World Health Organization


DANH MBNG

Trang
t s s
Bt
ng

Bt b s dng
Bt qu tng hlactat
Bng 3.2. Kt qu tng hp magie lactat b
3
ngay
khi b
Bng 3.3. Kt qu tng hp magie lactat b dng MgCO
3
khi
k
Bng 3.4. Kt qu tng hp magie lactat b sung dn MgCO
3

sau mi 24 gi
  ng sn ph    c b  

Bng 3.6. Kt qL. acidophilus  thu magie lactat  
Bng 3.7. Kt qu L. acidophilus  thu magie lactat v
n glucose
Bng MgCO
3
b sung sau mi 24 gi i n
u chnh pH = 7
Bc sau 96 gi i n
u chnh pH = 7

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DANH M  TH


Trang
L. acidophilus n vi quang hc
L. acidophilus n t

 magie lactat
 
c xut hi
 3.4. Ta trng xut hich NH
3

ch amoni clorid
ch trong such amoni clorid
t hin ta kt tinh trch Na
2
HPO
4

 t qu chy sp mng: sn phu ch c,
u ch bng h
hc (từ trái qua phải)
 u sut to magie lactat  
 u sut to magie lactat  
glucose
8
8
30
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1

T V
n pha vi sinh vc ng
dng ph bin trong mc ci sng. n phm c
ngh t, axit i lactat ng sn phi
t rt sng dng r Lactobacillus acidophilus c quan
u nhit trong nhng chng vi khu
y  p [4]nh canxi lactat vc nhc
te lactat t mui lactat ng dc
phi l ln [1], [8].
t cn thit cho s n th l ca
   magie  c t     p
Magie duy nh  nu
u ch t v quan trn thit.
agie lactat c s dng rt ph bing cha bnh,
magie lactat n sn phn xut canxi
lactat, axit   ng sn ph     s dng trong
p thc
phm, m ph [16], [22].
 git sn xut magie lactat 
mn.  Vi 
n, nhu v kh ng hp
u. Vi mong mu

cu sn xua ch 
u ch magie lactat bng h
Vi nhng m
-  nh o magie lactat.
- K.
2

. TNG QUAN

1.1. Magie lactat
1.1.1. m:
 c t
6
H
10
MgO
6
.
 c (theo IUPAC): Magie 2  hydroxyl propanoate.
 n: C 35,60%, H 4,98%, Mg 12,01%, O 47,42%.
 Kh: 202,45g/mol.
 c cu to: gm cation Mg
2+
 lactat.



Magie lactat i kt tinh ca axit lactic vi ion Mg
2+
, tn ti trong t

dng dihydrat (C
6
H
10
MgO
6
.2H
2
O) 
6
H
10
MgO
6
.3H
2
O).
 t: Magie lactat tn ti dng k
c lnh, tan tt trt tan trong
c ln 96
0
[23].

1.1.2. ng
Magie lactat c s dng trong nhic:
 Trong Y hc:
- t t n thi  h tr
cha h tim mch, th c
ng thng phi hp vi Vitamin B6.
- c s du tr thiu mc

s d  u tr chng       ng, h tr trong
chuyn ng c dng glucose.
3

- B sung mng h t do magie h tr
trong vip thu, t  
- Sn phm b sung magie, phi hp mng
s dng trong u tr thiu magie nng, thiu canxi lon thn kinh
ngou cht).
- S dng m gim nhng bin chi
i vi nhi nghin thu
- m s dng magie lactat b sung so vi m
loi mui mc s dt s v v hp thu
dn sinh kh d dng mui magie lactat ci thic v
i sinh kh di 99% [22].
  thc phm:
- t ph gia thc ph u v  t.
- Magie lactat  n phm b  
cht b  ung [16].
 c:
- Magie c s dc u vu chnh pH
- c trong d sn xut b
s i b y cao,
 th l gii th ci thip vi m
c s dc dp vic trong sn phm: sn
ph [22]
 p:
- Magie lactat n sn phn xut axit
i lactat i lactat. Axit n
xut cc phm, thc phm,

p dt, tng hp vecni, cht do, nhng nhit.
4

- Magie lactat c s d  tn
n t, thit b t  [22]

1.1.3. Mt s sn ph ng
t smagie lactat
STT



1
Maglek B6
Magie lactat,
Pyridoxin

Lek-AM, Poland Lek-AM,
Ba Lan
2
Magne B6
Sanofi-Aventis, Lithuania,
Sanofi-Aventis, Malta;
Sanofi-Aventis, Ba Lan
3
Magne B6 Stada
Stada-VN JV, Vietnam
Stada-VN JV, Vit Nam
4
Magne - B6

Corbière
Mekophar, Vit Nam

5
Magnelis B6

Nga
6
Magvit
GlaxoSmithKline, Ba Lan
7
Amfamag B6
Ampharco USA, Vit Nam
8
New - Mag
c
9
Magne-B6 Boston
Boston Pharma, Vit Nam
10
Magnerot
Magie lactat,
Axit orotic
a, Nga
11
Magnesium recip
Magie lactat,
Magie citrat

Recip, Thn

12
Magnesium
NYCOMED, Romania
13
Oromag
Theramex, Tunisia
14
Magnegon
Takeda Pharma, Th
15
Wamag
Magie lactat, hn
hp mui canxi
Hasco, Ba Lan

16
Magnesium
Glycocolle
Lafarge
Magie lactat,
glycin
SERP, Monaco


1.2. n xut magie lactat
1.2.1. ng hc
5

Bng h
cu tng h c magie lactat t      nhau 

trong sn xut axit lactic [14], [18].
- m:
+ D c magie lactat t nhiu ngu
 Th halogenid axit t axit  cloropropionic.
 Kh axit bng hydro m axit pyruvic.
 Cng h  ngun acetaldehyde [7].
 [27].
+ Thi gian ngn [27].
- m: Tng hp mng to
dng muaxit D (-axit racemic D,L - 
i vi sc khe c ci tin k thu
to sn phm taxit L (+) lactic v
cu [15].
-   ng r  o magie lactat  
tng hp cht ph gia thc phm, cht do, sn ph
[22].

1.2.2. t
 cu kin
k c vi hing vi sinh vt sinh axit 
      t nh  h enzym lactat dehydrogenase
chuyaxit lactic. Vic b p cht m
n dn hoc ngay t u s to ra mui magie
lactat ng thp tinh
thu d   thu sn phm.
6

Thc cht to magie lactat 
men to axit lactic.
   ng hp magie lactat t vi sinh v   

ng hng
m cc hi, d y, nhi sn xut thp,
ng sn xut thp, tn dn phm tha cc
phm, vn phm r c sn phm tinh khi
[21], [24].
- m:
+ To sn phm t  axit L (+) lactic [21].
+ Tn dt th
nghip sn xuo, ch bin sa, g [24].

1.3. Vi khun sinh axit lactic
1.3.1. n sinh axit lactic
Vi khun sinh lactic gc khun hay cu khun Gr (+), t l GC
) thp, chc axit 
hoc vi hia v
i chy
trong thc vn phm ca axit m chung ln
nht ca vi khu axit l
sn phi cht cu [3].
Vi khun sinh lactic ch yu thuc 4 chi: Lactobacillus, Pediococcus,
Streptococcus, Leuconostoc; t s m thuc chi Rhizopus
[1].
c ca vi khun sinh lactic rnc
tc mi.
7

a vi khun Gram
, hn y
hoc vi hi dnng t t ra axit lactic.
n ti tt  pH thp (4,5  6,8), nhi khong 10 

50
0
C [1], [3].
Vi khun d 
i phc t    ng, gm    axit  
peptid ng       u lo ng: glucose,
  dng
 [1].

1.3.2. Chi Lactobacillus
n nhn sinh lactic vi nhiu chc
ng dng trong sn xut. Chi Lactobacillus n vi
 chuy n phm to ra
 [3]:
-  n phm cua
 yaxit lactic (90 - n ph tn
ti dng vt nhi din: L. acidophilus, L. bulgaricus, L. delbrueckii…
-  t buc, sn phm
c axit lactic, axit acetic, axit 
i din: L. plantarum, L. casei, L. leichmenii…
-  t buc, sn phm cu
axit lactic, axit acetic, carbi din: L. brevis, L. fermentum,
L. buchneri…

1.3.3. Lactobacillus acidophilus
8

L. acidophilus thuc gii vi khuFermicutes, lp bacilli,
b lactobacilliales, h lactobacillaceae, ging lactobacillus [3], [6].

.1. L. acidophilus 
hin vi quang hc

 1.2. L. acidophilus  
hin t
L. acidophilus c khun Gram (+), dc
c to chui ngng, phn ng catalase
h; i khut, nhi t  45
0
C, chu
i thp (5  6); hp hi  y
 n ph [3],
[8].
L. acidophilus         i trong h
thng li vi khu vi
khui. L. acidophilus p t dch rut tr ch ru
Hin nay, L. acidophilus c s dng nhi ph
 u tr ng hp ri lo
tr mm ho [9].
L. acidophilus thun phm cui
axit c ng dng rrong thc tio qun thc
phm, dung dch v sinh ph n (D 

9

axit
- Ca axit lactic
c t
3
H
6
O
3
.
+ Kh: 90,08.
c cu to:



ng ca axit  ca axit 
baxit i quang: axit D (-) lactic, axit L (+) lactic.




Axit D (-) lactic Axit L (+) lactic
+ C





Axit D (-) lactic Axit L (+) lactic
Axit   t dng ch ng th     n hp

racemic. Dng axit racemic  t qu t 
tng h hp cht lactonitril hoc nh hong ca enzym DL
 lactatdehydrogenase do mt s  sinh v ng hp
axit lactic t [26].
10

axit t khi b 
d     chuy  ng axit    ng ca
enzym racemase t mn lactic.
+ t ca axit lactic:
Dng dung dch: axit t lng trong su
 
Dng tinh th: d tan  n to cht lng.
 pha lng: axit lactic d chuyn sang dng dimer mch thng lactoyl
ch th
  t ester v       ng dimer mch
 c thc hi
 140
0
t th b  quay cc.

1.4. u agie lactat
Peter Johannes Marie Baets ng s (2013)   
sn xut mui lactat b nguu thc vt,
m 25% cacbohydratt thc v
Mg(OH)
2
agie lactat [25].
Jan Van Krieken vs dng 
Mg(OH)

2
, MgO MgCO
3
duy 
cac 7 
maglactat magie t [20].
Gerrit Leendert Nanninga agie
lactat  d c t a cacaxit
c s d
 agie lactat. Magie
lactat kt tinh t      kh  t tinh
11

khit k ngun cacbohydrat  c s dng, thn
cacbohydrat  sucrose, tinh bt (lng [17].
Ben - Yoseph Eliahu, Kogan Leni, Wajc Samuel 
hu haxit lactic d 
cacbohydrat bi vi khun [11]. i phi kit ch
nhi  pH. M bin cho tt c 
c tit ra b S st
gii vi khuc s d 
i cht ca vi khung li. y, i ta ng
Ca(OH)
2
c canxi lactat, c
ngaxit  tinh khit cao vt m
 c s dng H
2
SO
4

 gi canxi lactat 
to ra canxi sult thi rn (th

agie lactat
 

2

 
canxi
lactate
 

      
 

axit
la clorua.
12

Sinh


Aharon Meir Eyal, Rod Fisher (1998) s

    Lactobacillus delbrueckii  Lactobacillus
acidophilus. libohydrat
 
              5,
t -   - 6,5

 -  
pH 
  
              
cacbonat, bicacbonat [10].

   MgO, Mg(OH)
2
 MgCO
3
     
(lact
nxi [20].
Bode Harold Eli           tinh ch
magie lactat t maxit ng, si
i b ta  magie sulfat  kt
t lactat, magie lactat; li
 l magie lactat kt tinh; 
 tinh khiagie lactat, ra vi c l[12].
Lesniak Wladyslaw, Przemysl R   MgCO
3
 
axit ng trng [19].
13

u v sinh tng hp, magie lactat c tng hp
b thc ph
    n xut magie
lactat t axit MgO u kin k c [27].
Mc s dng r

vin ti Vi 
n xut




















14

U

t liu, thit b
2.1.1. vt liu
 Chng ging: Lactobacillus acidophilus ATCC 4356 d  
(ngun gc: Vin Kim nghim).

 t
Bng 2.1. t

cht
Ngun gc

cht
Ngun gc
Glucose
Trung Quc
NaOH
Trung Quc
Sa bt
Vit Nam
NaK(C
4
H
4
O
6
)
Trung Quc
Cao nm men
c
H
2
SO
4
98%
Trung Quc

(NH
4
)
2
SO
4

Trung Quc
Na
2
S
2
O
3

Trung Quc
KH
2
PO
4
Trung Quc
KMnO
4
Trung Quc
MnSO
4
.H
2
O
Trung Quc

Br
2
Trung Quc
FeSO
4
.7H
2
O
Trung Quc
Isopropanol
Vit Nam
MgSO
4
.7H
2
O
Trung Quc
Methanol
Vit Nam
MgCO
3

Trung Quc
HCl
Trung Quc
CuSO
4
.5H
2
O

Trung Quc
NH
3
Vit Nam
Axit oxalic
Trung Quc
Br
2
Trung Quc
CuSO
4
.5H
2
O
Trung Quc
CuSO
4
.5H
2
O
Trung Quc
NaK(C
4
H
4
O
6
)
Trung Quc
HCl

Trung Quc
Na
2
S
2
O
3

Trung Quc
Isopropanol
Vit Nam
HCl
Vit Nam
Methanol
Vit Nam
Bn mng silica
gel G
c
NH
3
Vit Nam

15

 y s dng
 ng
ng


Glucose

4,0g
Sa bt
1,0g
Cao nm men
0,4g
(NH
4
)
2
SO
4

0,2g
KH
2
PO
4

0,25g
MnSO
4
.H
2
O
0,001g
FeSO
4
.7H
2
O

0,001g
MgSO
4
.7H
2
O
0,03g

100ml
u chnh pH = 6,3  6,5

 



Glucose
7,0g
Sa bt
2,6g
Cao nm men
1,3g
(NH
4
)
2
SO
4

0,2g
KH

2
PO
4

0,25g
MnSO
4
.H
2
O
0,001g
FeSO
4
.7H
2
O
0,001g
MgSO
4
.7H
2
O
0,03g

100ml
u chnh pH = 6,3  6,5


 ch s dng
ng dn c

 Dung d
c v 100 ml.
 Dung d       
4
.5H
2
  
ng 2-3 git H
2
SO
4
c v 500ml.
 Dung d         
NaK(C
4
H
4
O
6

2
 ngu H
2
O
v 500ml.
 Dung dch H
2
SO
4
     

2
SO
4
98% pha vi

2
i.
16

 Dung dch Na
2
S
2
O
3
     
2
S
2
O
3
 
Na
2
CO
3
c c
2

2

O v 1000ml.
 Dung dch HCl 0,0axit hydrocloric vc va
 100 ml.

2.1.2. Thit b s dng
Bng 2.4. t b s dng
Thit b, dng c
Ngun gc, xut x
thut
c

c
Ni hp
ALP, Nht
T cy
Sanyo, Nht
T m
c
T sy
c

c
nh
c
Bp y
HHS, Trung Quc
T lnh
Daewoo, c

c

nh mc, pipet, ng nghing c
pha ch

2.2. Nu
2.2.1.  ng o magie lactat
 La chn agie cacbonat
- B
- 
17

- K
 La chn thu sn phm magie lactat
- Lng phu lc Buchner.
- ng.
- nh.
  sn phm magie lactat
- 
2+
.
- 
- Sp mng.
2.2.2. Kht s  
 Khng ca pH: 6  8.
 Khng ca n glucose: 7  12%.

u
ging:
  ng  .2)  c tit   0,6 atm
  ngui xung 40  45
0

C s cy chng L. acidophilus. 
trong t m  nhi 37
0
C. Sau 24 gi c ging cn thit.
Nu gic gi  nhi thp (trong t li
 ng v tr   ng b  trong t m nhi  37
0
C
trong 2 - i ti [7].

2.3.2.ng hp magie lactat:
Chun b 2.3) vi c
l            u ti
ng  0,6
 ngun nhi khong 40  45
0
C, trn l