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Biocellulose – A new type of separation membrane
Biocellulose Membrane Biocellulose Membrane
A. Rudloff*, M. Winter, S. Kreusch, R. Rubick, K. Frankenfeld* S c i e n o va G m b H , S p i t z w e i d e nw e g 3 0 , 0 7 7 4 3 J e n a , G e r m a ny, * fzmb GmbH, Geranienweg 7, 99947 Bad Langensalza, Germany
scienova GmbH Spitzweidenweg 30,07743 Jena, Germany
T +49 (0)3641 504 586 F +49 (0)3641 504 587
www.scienova.com [email protected]
Biocellulose Biocellulose
Biocellulose for Dialysis Biocellulose for Dialysis
Methods Methods Fig 9 and 10 Growth All bacterias were cultivated at 37 °C and 225 rpm. Colonies were picked from the agar plates and cultivated overnight. The overnight culture was diluted in fresh medium. The growth was monitored by determining the optical densitiy at 600 nm with a nanophotometer. To compare the efficiency of the bioreactor all experiments were done in the BR1000 (1 ml) as well as in the Erlenmeyer flask (50 ml) .
I
II
III
This work is supported by the BMWI (Federal Department of Commerce of Germany), KF2937502 CR4
The Biocellulose membrane has a 3 D
fiber structure and consists of cellulose
nanofibres. Biocellulose is synthesized
by Acetobacter. It is mechanical and
chemical stable.
The first available Biocellulose
membrane is autoclavable, it is
biocompatible, has protein binding
<10 µg/cm2 of BSA and is storable as a
dried membrane.
Fig.1 Cellulose nanofibres of the Bio-
cellulose membrane (Source: fzmb GmbH) Fig. 2 Acetobacter (Source: fzmb GmbH)
Fig. 3 Wet Biocellulose from batch
(Source: fzmb GmbH)
Fig. 4 Heat dried Biocellulose sheets
(Source: fzmb GmbH)
b b
Biocellulose for Cell Culture Biocellulose for Cell Culture
0
20
40
60
80
100
10 100 1000
Re
nte
nti
on
[%
]
log Molekulargewicht [kDa]
Biocellulose cut off 140 kDa can be
combined with dialysis devices from
scienova compatible to micro plate
format. The dialysis efficiency is in the
same range as for regenerated cellulose.
Fig. 5 ED300 (Xpress Dialyzer
Family) with Biocellulose
membrane into a 96 well
Riplate (Source: scienova GmbH)
Fig. 6 Sieve curve
determined the cut off
140 kDa
0
10
20
30
40
50
60
70
80
90
100
-40 10 60 110 160
Par
anit
rop
he
no
le c
on
c. in
%
t in min
RC ED300
RC DWP
BC ED300
BC DWP
Fig. 7 Equilibrium Dialysis of
Paranitrophenole. ED300 with
BC Biocellulose and RC
regenerated Cellulose
Fig. 9 gram-negative bacteria Fig. 10 gram-positive bacteria
0,01
0,10
1,00
10,00
0 2 4 6 8
log
OD
[6
00
nm
]
t in h
Bacillus licheniformis
BR1000
Erlenmeyerkolben
0,01
0,10
1,00
10,00
0 2 4 6 8
log
OD
[6
00
nm
]
t in h
Lactobacillus jensenii
BR1000
Erlenmeyerkolben
0,01
0,10
1,00
10,00
0 2 4 6 8
log
OD
[6
00
nm
]
t in h
Listeria monocytogenes
BR1000
Erlenmeyerkolben
0,01
0,10
1,00
10,00
0 2 4 6 8
log
OD
[6
00
nm
]
t in h
Streptococcus agalactiae
BR1000
Erlenmeyerkolben
0,01
0,10
1,00
10,00
0 2 4 6 8 10
log
OD
[6
00
nm
]
t in h
Klebsiella pneumoniae
BR1000
Erlenmeyerkolben
0,01
0,10
1,00
10,00
0 2 4 6 8 10
log
OD
(6
00
nm
)
t in h
Pseudomonas aeruginosa
BR1000
Erlenmeyerkolben
0,01
0,10
1,00
10,00
0 2 4 6 8
log
OD
[6
00
nm
]
t in h
Legionella pneumophilia
BR1000
Erlenmeyerkolben
0,01
0,10
1,00
10,00
0 2 4 6 8
log
OD
[6
00
nm
t in h
E.coli
BR1000
Erlenmeyerkolben
Biocellulose is known as biocompatible. A BR1000 with
Biocellulose membrane was tested as membrane reactor for
the culture of gram-negative and -positive bacteria compared
with usual Erlmeyer flask culture. The growth curve
progression was mostly similar. Cell density was lower. The
reason could be a lower oxygen maintenance.
Fig. 8 a) single BR1000, b) BR1000 into 48 well plate, c) BR1000 with Biocellulose
membrane in 48 well Riplate
a b c
Fig 6 Sieve curve: Microdialyzer were filled with 1 ml of the sample and placed in 2.7 ml of the dialyses buffer at room temperature. The solutions were not changed during the whole process. Dialyses buffer was not adjusted. Determination of retention after 48 h dialysis. Optical density were measured by nanoDrop. (test
substances: fluerescence particle (700 kDa), IgG (150 kDa), bovine serum albumin (67 kDa), Streptavidin (60 kDa), horseradish peroxidase (44 kDa), Casein (20 kDa)
Fig 7 Equilibrium: scienova Xpress Equilibrium Dialyzer ED300 either with regenerated cellulose 6-8 kDa cut off or with Biocellulose 140 kDa, each were filled with 300 µl 1mM Paranitrophenole in PBS and dialysed against 1.2 ml PBS pH7.4 in a 96 well Riplate at ambient temperature. Measurement of absorbance were performed at 420 nm (ref. 620 nm).
Video demonstration:
scienova Xpress Equilibrium Dialyzer ED300 and
Analytik Jena Cybio® FeliX
