Novel cross-linked alcohol-insoluble solid (CL-AIS) affinity gel from pea pod for pectinesterase purification.
Alcohol-insoluble solids (AIS) from pea pod were cross-linked (CL-AIS) and used as an affinity gel matrix to isolate pectin esterases (PEs) from tendril shoots of chayote (TSC) and jelly fig achenes (JFA), and the results were compared with those isolated by ion-exchange chromatography with a commercial resin. CL-AIS gel matrix in a column displayed poor absorption and purification fold of PE; however, highly methoxylated CL-AIS (HM-CL-AIS), by exposing CL-AIS to methanolic sulfuric acid to increase the degree of esterification (DE) to 92%, facilitated the enzyme purification. The purified TSC PE and JFA PE by the HM-CL-AIS column were proofed as a single band on an SDS-PAGE gel, showing that the HM-CL-AIS column was a good matrix for purification of PE, either with alkaline isoelectric point (pI) (TSC PE) or with acidic pI (JFA PE).
Wu
Ming-Chang
MC
Department of Food Science, National Pingtung University of Science and Technology, Pingtung 912-07, Taiwan. Changm@ntu.edu.tw
Lin
Guan-Hui
GH
Wang
Yuh-Tai
YT
Jiang
Chii-Ming
CM
Chang
Hung-Min
HM
D000438:Alcohols; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D003432:Cross-Linking Reagents; D019662:Cucurbitaceae / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004951:Esterification; D030681:Ficus / Q000201:enzymology; D005782:Gels / Q000737:chemistry*; D018532:Pisum sativum / Q000737:chemistry*; D012995:Solubility
D000438:Alcohols; D003432:Cross-Linking Reagents; D005782:Gels; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase