title: Novel cross-linked alcohol-insoluble solid (CL-AIS) affinity gel from pea pod for pectinesterase purification.
abstract: 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).
authors.affiliation: Department of Food Science, National Pingtung University of Science and Technology, Pingtung 912-07, Taiwan. Changm@ntu.edu.tw
authors.forename: Ming-Chang
authors.initials: MC
authors.lastname: Wu
authors.forename: Guan-Hui
authors.initials: GH
authors.lastname: Lin
authors.forename: Yuh-Tai
authors.initials: YT
authors.lastname: Wang
authors.forename: Chii-Ming
authors.initials: CM
authors.lastname: Jiang
authors.forename: Hung-Min
authors.initials: HM
authors.lastname: Chang
mesh_terms: 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
chemical_list: D000438:Alcohols; D003432:Cross-Linking Reagents; D005782:Gels; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase