Nghiên cứu một số hướng ứng dụng màng BC tạo ra từ vi khuẩn gluconacetobacter


KHOA SINH - KTNN
======







GLUCONACETOBACTER










2014


KHOA SINH - KTNN
======







GLUCONACETOBACTER


 c

:
















 TS.   ,        
ng d
t - Vi sinh, khoa Sinh - 
 

m khoa Sinh - 
 
 .



Hà Nội, ngày 12 tháng 4 năm 2014
Sinh viên














 m,              
         


Hà Nội, ngày 12 tháng 4 năm 2014
Sinh viên





 1

 1
 2
 2
 2
 2
 2
.  3
          Gluconacetobacter trong
 3
Gluconacetobacter 3
Gluconacetobacter 5
Gluconacetobacter 7
 7
1. 8
 8
 9
a cellulose và màng BC 9
 10
 11
 11
 11
.  13
 13
 13
 13
 14
 14
2. 15
 16
 16

 17
.  18
3.1. QGluconacetobacter BHN
2
18
 
Gluconacetobacter BHN
2
18
Gluconacetobacter
BHN
2
19
 20
 20
 23
 25
 29
 29
 29
 30



 

A.xylinum
Acetobacter xylinum
ASTM
American society for testing and materials

BC
Bacterial cellulose
CS
Cellulose synthate
Cs

G
-

Gram âm
GK
Glucokinase
Nxb

PMG
Phosphoglucomutase
TCVN

TP.HCM

UDPGlc
Uridine diphospho glucose
UGP
Glucose-1-phosphate uridylytransferase



 4
Gluconacetobacter 7
 21
 23

 25




Hình 1.1. Gluconacetobacter 11
        Gluconacetobacter BHN
2
 
       Gluconacetobacter BHN
2
trên
MT1 19
Hình 3.2. Màng BC sau 5 ngày lên men 20
 20
 22
 22
 22
 24
 24
 24
Hình 3.10. Nem chua b 24

 24
 27
Hình 3.13. Táo sau 5 ngày 27
 27
 27
 27
 27
 28
 28
 28
 28
Hình 3. 28
 28
Nguyễn Thị Thùy 1

M U

1. Lý do ch tài
c (Bacterial cellulose; Biocellulose; BC

      -      
  igin,
hemicellulose, pept 

Bacterial cellulose 
 khác nhau: N
 
 bào, 
           
 quá trình

ch,
 [21], [22], [24].

           
           
 
       
 
 2 phân , Gluconacetobacter BHN
2


“Nghiên cứu một số hướng
ứng dụng màng BC tạo ra từ vi khuẩn Gluconacetobacter”.
Nguyễn Thị Thùy 2

2. Mc tiêu c tài


3. Ni dung
Gluconacetobacter
BHN
2



c và thc tin
4.1. Ý nghĩa khoa học: n vi 

4.2. Ý nghĩa thực tiễn:          

m và 
4.3. Điểm mới
Gluconacetobacter BHN
2
 
600cm
2
, 1300cm
2

.
Nguyễn Thị Thùy 3


TNG QUAN TÀI LIU

1.1. V  m phân loi ca chng Gluconacetobacter trong
sinh gii
1.1.1. Vị trí phân loại của chủng Gluconacetobacter

1898, Beijerinck 1898, Hoyer 1899, Hansen 1911, Henenberg 1926,
  Gluconacetobacter
 Acetobacter khi Yamada và
          
- 10 là thành
n ubGluconacetobacter,
trong khi Q - n ub
Acetobacter     Gluconacetobacter    

              

Gluconacetobacter 
Gluconacetobacter nom. corrig, theo 
      Gluconacetobacter  


     Gluconacetobacter     
             
Acetobacter Gluconacetobacteri cùng Yamada (2000)
        Gluconacetobacter    
ng t là
Nguyễn Thị Thùy 4

D - u Q - n ub
            
Gluconacetobacter           


AND.
Theo Bergey (2005) [16], G.intermedius    
Gluconacetobacter 
Acetobacter, Acidomonas, Asaia, Gluconobacter, Gluconacetobacter và
Kozakia


(1)
(2)
(3)
(4)
(5)
(6)

Oxy hóa ethanol thành acid
acetic
+
+
-/w
+
+
+
Oxy hóa acid acetic thành
CO
2
và H
2
O
+
+
+
+
-
w
Oxy hóa lactate thành CO
2
và
H
2
O
+
w
+
+/-

-
w



+

+

-

+

+

+
Sinh  - manitol
+/-
w
+/-
+/-
+
+

-/w
+
-
_
-
-


-
-
-
+/-
-
-
Oxy hóa glixerol thành
dihydroxyaceton
+/-
w
-/w
+/-
+
+







Nguyễn Thị Thùy 5

D-manitol
Glycerol
Raffinose
-/+
-/+
-

-
-
Nd
+/-
+
-
+/-
+
-
+
+
-
-
+
+
i ubiqiunone
Q-10
Q-10
Q-10
Q-10
Q-10
Q-10
%mol G + C
52-60
63-66
59-61
55-66
54-63
56-57

Chú thích: (1): Acetobacter (4): Gluconacetobacter +: kết quả dương tính
(2): Acidomonas (5): Gluconobacter -: kết quả âm tính
(3): Asaia (6): Kozakia nd: Chưa xác định
w: yếu

1.1.2. Đặc điểm phân loại của chủng Gluconacetobacter
Acetobacter xylinum BHN
2
n Thùy Vân

am và cs [1], [3],
 A.
xylinum BHN
2
trên (

 
             
BHN
2
Gluconacetobacter intermedius
AB 166739, Gluconacetobacter intermedius AB099296,
Gluconacetobacter intermedius 
   Gluconacetobacter BHN
2
_21 là Gluconacetobacter
intermedius [1].


Nguyễn Thị Thùy 6


CGluconacetobacter 


H
2
SO
4

 c           

.

      Gluconacetobacter hình thành


Gluconacetobacter g

BC. 

 
.

 m sinh lý: C Gluconacetobacter   pH: 4 - 6,
- 35
o

o


. Gluconacetobacter 
         

 N
thành
CO
2
và H
2
          
Nguyễn Thị Thùy 7


ng 1.2:
Gluconacetobacter
STT




1
Oxy hoá ethanol thành
acid acetic


+
2


+

3
   


_
4
  
thành dihydroxyaceton

+
5
  
thành acid


3

+
6
    


_
7
    


+

1.2. Nhu cu ng ca chng Gluconacetobacter


            
       
4
, (NH
4
)
2
SO
4
, (NH
4
)
2
HPO
4
,
MgSO
4
. 7H
2
O, KH
2
PO
4

1.2.1. Nhu cầu nguồn cacbon

              
 

           
Nguyễn Thị Thùy 8

             
  
c 0,5 atm 110
o
C 
ng dùng 0,2 - 
.
1.2.2. Nhu cầu nguồn nitơ
(NH
4
)
2
SO
4

    Gluconacetobacter.    
Gluconacetobacter   
4
+
 
NH
4
+

4
2-
, Cl

-

Gluconacetobacter 

              
. 

Gluconacetobacter.


.
1.2.3. Nhu cầu nguồn phospho



2
PO
4
, K
2
HPO
4
, KNO
3



    Gluconacetobacter và quá trình hình thành màng

Nguyễn Thị Thùy 9

Gluconacetobacter 

1.3.  hình thành màng BC
1.3.1. Đặc điểm cấu trúc của cellulose và màng BC
1.3.1.1. Đặc điểm cấu trúc của cellulose

5
- 22,68.10
5


6
H
10
O
5
)
n

 

-               
glucose. Cellul
        a các nhóm (- 

c monosaccharide. Cellulose có tính

        
nhau.



o


             
     
3
       
2
,
PbCl
2

        -    
    -        
lucose.
(C
6
H
10
O
5
)
n
+ nH
2
O -> nC
6
H

12
O
6

Nguyễn Thị Thùy 10



[C
6
H
7
O
2
(OH)
3
]
n
+ 3n HNO
3
-> [C
6
H
7
O
2
(ONO
2
)
3

]
n
+ 3nH
2
O
1.3.1.2. Đặc điểm cấu trúc của màng BC
-
 -
subfibril
- 
 




         




1.3.2. Cơ chế tổng hợp màng BC




    Gluconacetobacter BHN2: Glucokinase (GK),
Phosphoglucomutase (PGM), Glucose-1-phosphate uridylyltransferase
(UDP

Nguyễn Thị Thùy 11

Hình 1.1. Con đường sinh tổng hợp cellulose ở chủng Gluconacetobacter
1.4. Tình hình nghiên cu màng BC
1.4.1. Tình hình nghiên cứu màng BC trên thế giới
Gluconacetobacter



Gluconacetobacter 
           m. Trong công



dùng l

 

1.4.2. Tình hình nghiên cứu màng BC ở Việt Nam
            
Gluconacetobacter 
Gluconacetobacter 


Nguyễn Thị Thùy 12



 N

   A. xylinum          .
 - 
 
 
 Ðinh 
Gluconacetobacter BHN
2

bao bì


Nguyễn Thị Thùy 13


U

ng nghiên cu và hóa cht
2.1.1. Đối tượng nghiên cứu
 Gluconacetobacter BHN
2

 phòng Vi Sinh, Khoa sinh - 
2.1.2. Hoá chất và thiết bị
2.1.2.1. Hoá chất

n cacbon: Ethanol, Glucose, Acid acetic, Agar.

4
)
2
SO
4
.
KH
4
PO
4
, CaCO
3
, MgSO
4
.7H
2
O.


ng: N
2.1.2.2. Thiết bị


Shakergallenkump (Anh).

 20µl - 10ml.

Cân (Precisa XT 320M - 


 


Nguyễn Thị Thùy 14

2.1.2.3. Môi trường nghiên cứu

MT1
MT2
MT3
MT4
MT5
1. Glucose
20g
20g
20g
20g
20g
2. Agar
20g
0
0
0
0
3. (NH
4
)
2
SO

4
3g
3g
3g
5g
3g
4. KH
2
PO
4
2g
2g
2g
2g
2g
5. MgSO
4
.7H
2
O
2g
2g
2g
0
2g
6. Acid acetic
2%
2%
2%
5ml

2.5ml
etylic
2%
2%
2%
0
2%
8. Pepton
0
5g
0
0
5g

0
0
0
0
1000ml

1000ml
1000ml
1000ml
1000ml
0


.
Tr


MT3 
u
2.2.1. Phương pháp vi sinh
2.2.1.1. Phương pháp phân biệt tế bào chủng Gluconacetobacter bằng
phương pháp nhuộm Gram
Gluconacetobacter   
           
   Gluconacetobacter      


Nguyễn Thị Thùy 15

2.2.1.2. Phương pháp bảo quản chủng Gluconacetobacter trên môi trường
thạch nghiêng
           
Gluconacetobacter  - 5 ngày trong

o
C. Sau 
o
C 
p 

2.2.1.3. Phương pháp hoạt hóa giống


             
ch a
o
C trong



2.2.1.4. Phương pháp lên men tạo màng BC

o
C
 

 - 8 ngày [1], [9].
2.2.1.5. Phương pháp xử lí màng BC sinh từ vi khuẩn Gluconacetobacter
2.2.2. Phương pháp vật lí
2.2.2.1. Phương pháp xác định dai (độ bền cơ học) của màng BC

             
 

828 - 93 (American society for
Nguyễn Thị Thùy 16

tessting and materials) TCVN 1862 - y và
xenlulo.
2.2.3 Phương pháp hóa học
2.2.3.1. Phương pháp làm trắng, làm sạch chất dư thừa trong quá trình
nuôi cấy
i John Mercer (1850) - 

m. H
 

2.2.3.2. Phương pháp chuẩn độ pH của màng

             
m


2.2.4. Phương pháp bảo quản ứng dụng
2.2.4.1. Phương pháp bảo quản màng BC sinh từ vi khuẩn
Gluconacetobacter




2.2.4.2. Phương pháp ứng dụng làm bao bì bảo quản thực phẩm
m: D
: T 
ng 
