Continuous purification of Candida antarctica lipase B using 3‐membrane adsorber periodic counter‐current chromatography
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 wa...
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crwiley:10.1002/elsc.201700159 2024-06-02T07:58:26+00:00 Continuous purification of Candida antarctica lipase B using 3‐membrane adsorber periodic counter‐current chromatography Brämer, Chantal Schreiber, Sarah Scheper, Thomas Beutel, Sascha Bundesministerium für Bildung und Forschung 2018 http://dx.doi.org/10.1002/elsc.201700159 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Felsc.201700159 https://onlinelibrary.wiley.com/doi/pdf/10.1002/elsc.201700159 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Engineering in Life Sciences volume 18, issue 7, page 414-424 ISSN 1618-0240 1618-2863 journal-article 2018 crwiley https://doi.org/10.1002/elsc.201700159 2024-05-03T11:00:02Z 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/cm 2 ). 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. Article in Journal/Newspaper Antarc* Antarctica Wiley Online Library Engineering in Life Sciences 18 7 414 424 |
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Wiley Online Library |
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English |
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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/cm 2 ). 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. |
author2 |
Bundesministerium für Bildung und Forschung |
format |
Article in Journal/Newspaper |
author |
Brämer, Chantal Schreiber, Sarah Scheper, Thomas Beutel, Sascha |
spellingShingle |
Brämer, Chantal Schreiber, Sarah Scheper, Thomas Beutel, Sascha Continuous purification of Candida antarctica lipase B using 3‐membrane adsorber periodic counter‐current chromatography |
author_facet |
Brämer, Chantal Schreiber, Sarah Scheper, Thomas Beutel, Sascha |
author_sort |
Brämer, Chantal |
title |
Continuous purification of Candida antarctica lipase B using 3‐membrane adsorber periodic counter‐current chromatography |
title_short |
Continuous purification of Candida antarctica lipase B using 3‐membrane adsorber periodic counter‐current chromatography |
title_full |
Continuous purification of Candida antarctica lipase B using 3‐membrane adsorber periodic counter‐current chromatography |
title_fullStr |
Continuous purification of Candida antarctica lipase B using 3‐membrane adsorber periodic counter‐current chromatography |
title_full_unstemmed |
Continuous purification of Candida antarctica lipase B using 3‐membrane adsorber periodic counter‐current chromatography |
title_sort |
continuous purification of candida antarctica lipase b using 3‐membrane adsorber periodic counter‐current chromatography |
publisher |
Wiley |
publishDate |
2018 |
url |
http://dx.doi.org/10.1002/elsc.201700159 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Felsc.201700159 https://onlinelibrary.wiley.com/doi/pdf/10.1002/elsc.201700159 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Engineering in Life Sciences volume 18, issue 7, page 414-424 ISSN 1618-0240 1618-2863 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1002/elsc.201700159 |
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Engineering in Life Sciences |
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18 |
container_issue |
7 |
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414 |
op_container_end_page |
424 |
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1800741770299965440 |