TY - JOUR
T1 - Separation of active laccases from Pleurotus sapidus culture supernatant using aqueous two-phase systems in centrifugal partition chromatography
AU - Schwienheer, C.
AU - Prinz, A.
AU - Zeiner, T.
AU - Merz, J.
PY - 2015/10/1
Y1 - 2015/10/1
N2 - For the production of bio active compounds, e.g., active enzymes or antibodies, a conserved purification process with a minimum loss of active compounds is necessary. In centrifugal partition chromatography (CPC), the separation effect is based on the different distribution of the components to be separated between two immiscible liquid phases. Thereby, one liquid phase is kept stationary in chambers by a centrifugal field and the mobile phase is pumped through via connecting ducts. Aqueous two phase systems (ATPS) are known to provide benign conditions for biochemical products and seem to be promising when used in CPC for purification tasks. However, it is not known if active biochemical compounds can "survive" the conditions in a CPC where strong shear forces can occur due to the two-phasic flow under centrifugal forces. Therefore, this aspect has been faced within this study by the separation of active laccases from a fermentation broth of Pleurotus sapidus. After selecting a suitable ATPS and operating conditions, the activity yield was calculated and the preservation of the active enzymes could be observed. Therefore, CPC could be shown as potentially suitable for the purification of bio-active compounds.
AB - For the production of bio active compounds, e.g., active enzymes or antibodies, a conserved purification process with a minimum loss of active compounds is necessary. In centrifugal partition chromatography (CPC), the separation effect is based on the different distribution of the components to be separated between two immiscible liquid phases. Thereby, one liquid phase is kept stationary in chambers by a centrifugal field and the mobile phase is pumped through via connecting ducts. Aqueous two phase systems (ATPS) are known to provide benign conditions for biochemical products and seem to be promising when used in CPC for purification tasks. However, it is not known if active biochemical compounds can "survive" the conditions in a CPC where strong shear forces can occur due to the two-phasic flow under centrifugal forces. Therefore, this aspect has been faced within this study by the separation of active laccases from a fermentation broth of Pleurotus sapidus. After selecting a suitable ATPS and operating conditions, the activity yield was calculated and the preservation of the active enzymes could be observed. Therefore, CPC could be shown as potentially suitable for the purification of bio-active compounds.
KW - Activity preservation
KW - Aqueous two-phase systems (ATPS)
KW - Centrifugal partition chromatography (CPC)
KW - Downstream processing
KW - Laccase from Pleurotus sapidus
UR - http://www.scopus.com/inward/record.url?scp=84939614388&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2015.07.050
DO - 10.1016/j.jchromb.2015.07.050
M3 - Article
C2 - 26295695
AN - SCOPUS:84939614388
SN - 1570-0232
VL - 1002
SP - 1
EP - 7
JO - Journal of Chromatography B
JF - Journal of Chromatography B
ER -