Extraction of peptide tagged cutinase in detergent-based aqueous two-phase systems.
Detergent-based aqueous two-phase systems have the advantage to require only one auxiliary chemical to induce phase separation above the cloud point. In a systematic study the efficiency of tryptophan-rich peptide tags was investigated to enhance the partitioning of an enzyme to the detergent-rich phase using cutinase as an example. Up to 90% enzyme activity could be extracted in a single step from whole broth of recombinant Saccharomyces cerevisiae expressing cutinase variants carrying a (WP)4 tag. In contrast, the extraction yield of wild type cutinase was 2-3% only. The detergent concentration and the temperature are the main parameters to optimize the extraction yield. Considering availability, extraction yields, and price the detergent Agrimul NRE 1205 served best for enzyme recovery.
Rodenbrock
A
A
Institute of Enzyme Technology, Heinrich-Heine University, Jülich, Germany.
Selber
K
K
Egmond
M R
MR
Kula
M R
MR
D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003902:Detergents; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification; D009842:Oligopeptides / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D014867:Water
D003902:Detergents; D005656:Fungal Proteins; D009842:Oligopeptides; D011994:Recombinant Proteins; D014867:Water; D002265:Carboxylic Ester Hydrolases; C019952:cutinase