TY - JOUR
T1 - Vortex fluidic device-intensified aqueous two phase extraction of C-phycocyanin from Spirulina maxima
AU - Luo, Xuan
AU - Smith, Paul
AU - Raston, Colin
AU - Zhang, Wei
PY - 2016/7/5
Y1 - 2016/7/5
N2 - A vortex fluidic device (VFD) is effective for downstream processing of C-phycocyanin (C-PC) from blue-green microalga Spirulina maxima using an aqueous two phase system (ATPS): polyethylene glycol (PEG) and aqueous potassium phosphate. The VFD-intensified ATPS continuous flow processing (VFD-ATPS) has spontaneous phase separation of the liquid exiting the 20 mm diameter tube inclined at 45° and rotating at 6550 rpm, for a 0.2 mL/min flow rate. This processing results in at least a 9.3-fold increase in phase demixing efficiency, 1.18-fold increase in C-PC purity, and 22% increase in yield compared to conventional ATPS methods. Multistage counter current distribution (CCD) using VFD-ATPS followed by removal of PEG and salt increased C-PC purity by 6-fold compared to the crude extract, with triple TOF mass spectrometry showing more than 95% sequence coverage for both subunits. This work establishes the utility of using VFD-intensified ATPS processing for protein purification and its potential as an effective new tool for rapid processing of biologically active and intact proteins.
AB - A vortex fluidic device (VFD) is effective for downstream processing of C-phycocyanin (C-PC) from blue-green microalga Spirulina maxima using an aqueous two phase system (ATPS): polyethylene glycol (PEG) and aqueous potassium phosphate. The VFD-intensified ATPS continuous flow processing (VFD-ATPS) has spontaneous phase separation of the liquid exiting the 20 mm diameter tube inclined at 45° and rotating at 6550 rpm, for a 0.2 mL/min flow rate. This processing results in at least a 9.3-fold increase in phase demixing efficiency, 1.18-fold increase in C-PC purity, and 22% increase in yield compared to conventional ATPS methods. Multistage counter current distribution (CCD) using VFD-ATPS followed by removal of PEG and salt increased C-PC purity by 6-fold compared to the crude extract, with triple TOF mass spectrometry showing more than 95% sequence coverage for both subunits. This work establishes the utility of using VFD-intensified ATPS processing for protein purification and its potential as an effective new tool for rapid processing of biologically active and intact proteins.
KW - Aqueous two phase system
KW - C-Phycocyanin
KW - Microalgae
KW - Phase demixing
KW - Vortex fluidics device
UR - http://www.scopus.com/inward/record.url?scp=84978974789&partnerID=8YFLogxK
U2 - 10.1021/acssuschemeng.6b00756
DO - 10.1021/acssuschemeng.6b00756
M3 - Article
SN - 2168-0485
VL - 4
SP - 3905
EP - 3911
JO - ACS Sustainable Chemistry & Engineering
JF - ACS Sustainable Chemistry & Engineering
IS - 7
ER -