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 -