Details
| Original language | English |
|---|---|
| Pages (from-to) | 414-424 |
| Number of pages | 11 |
| Journal | Engineering in life sciences |
| Volume | 18 |
| Issue number | 7 |
| Early online date | 2 Apr 2018 |
| Publication status | Published - 8 Jul 2018 |
Abstract
Batch chromatography has several disadvantages, such as insufficient utilization of the capacity of the resin, high buffer consumption and discontinuity. Considering the high costs for downstream processing, a continuously working chromatographic system with three membrane adsorber units was designed, tested and put into operation. The basic principle of the setup is periodic counter-current chromatography (PCCC). The PCCC system was used for capturing and purifying Candida antarctica lipase B (CalB) directly from cell lysate in one single unit operation. The best purification result was achieved by means of anion-exchange chromatography. The dynamic binding capacity with Sartobind® Q 75 amounted to 4.2 mg (56 g/cm2). After transferring the method to the 3MA-PCCC, 0.22 g CalB (73 U/mg) were obtained from 0.9 L E. coli lysate within 6 h and a recovery of 80%. Compared to the batch process, the productivity could be increased by 36% and the buffer consumption could be reduced by about 20%. Although the purification of CalB from lysate by means of anion-exchange chromatography was not selective and quantitative using the 3MA-PCCC device, it could be shown that the concept of the system was successfully implemented and led to a significant improvement of CalB purification.
Keywords
- Candida antarctica lipase B, Continuous chromatography, Membrane adsorber, Multi column chromatography, Periodic counter-current chromatography
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Environmental Science(all)
- Environmental Engineering
- Chemical Engineering(all)
- Bioengineering
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In: Engineering in life sciences, Vol. 18, No. 7, 08.07.2018, p. 414-424.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Continuous purification of Candida antarctica lipase B using 3-membrane adsorber periodic counter-current chromatography
AU - Brämer, Chantal
AU - Schreiber, Sarah
AU - Scheper, Thomas
AU - Beutel, Sascha
N1 - Funding information: The studies were carried out within the frame of project P38 of the BMBF-Biokatalyse2021-Cluster, hosted by Prof. Garo Antranikian. We would like to thank the German Federal Ministry of Education and Research for their financial support.
PY - 2018/7/8
Y1 - 2018/7/8
N2 - Batch chromatography has several disadvantages, such as insufficient utilization of the capacity of the resin, high buffer consumption and discontinuity. Considering the high costs for downstream processing, a continuously working chromatographic system with three membrane adsorber units was designed, tested and put into operation. The basic principle of the setup is periodic counter-current chromatography (PCCC). The PCCC system was used for capturing and purifying Candida antarctica lipase B (CalB) directly from cell lysate in one single unit operation. The best purification result was achieved by means of anion-exchange chromatography. The dynamic binding capacity with Sartobind® Q 75 amounted to 4.2 mg (56 g/cm2). After transferring the method to the 3MA-PCCC, 0.22 g CalB (73 U/mg) were obtained from 0.9 L E. coli lysate within 6 h and a recovery of 80%. Compared to the batch process, the productivity could be increased by 36% and the buffer consumption could be reduced by about 20%. Although the purification of CalB from lysate by means of anion-exchange chromatography was not selective and quantitative using the 3MA-PCCC device, it could be shown that the concept of the system was successfully implemented and led to a significant improvement of CalB purification.
AB - Batch chromatography has several disadvantages, such as insufficient utilization of the capacity of the resin, high buffer consumption and discontinuity. Considering the high costs for downstream processing, a continuously working chromatographic system with three membrane adsorber units was designed, tested and put into operation. The basic principle of the setup is periodic counter-current chromatography (PCCC). The PCCC system was used for capturing and purifying Candida antarctica lipase B (CalB) directly from cell lysate in one single unit operation. The best purification result was achieved by means of anion-exchange chromatography. The dynamic binding capacity with Sartobind® Q 75 amounted to 4.2 mg (56 g/cm2). After transferring the method to the 3MA-PCCC, 0.22 g CalB (73 U/mg) were obtained from 0.9 L E. coli lysate within 6 h and a recovery of 80%. Compared to the batch process, the productivity could be increased by 36% and the buffer consumption could be reduced by about 20%. Although the purification of CalB from lysate by means of anion-exchange chromatography was not selective and quantitative using the 3MA-PCCC device, it could be shown that the concept of the system was successfully implemented and led to a significant improvement of CalB purification.
KW - Candida antarctica lipase B
KW - Continuous chromatography
KW - Membrane adsorber
KW - Multi column chromatography
KW - Periodic counter-current chromatography
UR - http://www.scopus.com/inward/record.url?scp=85046106571&partnerID=8YFLogxK
U2 - 10.1002/elsc.201700159
DO - 10.1002/elsc.201700159
M3 - Article
C2 - 32624922
AN - SCOPUS:85046106571
VL - 18
SP - 414
EP - 424
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 7
ER -