title: Biomimetic affinity purification of Candida antarctica lipase B.
abstract: Candida antarctica lipase B (CalB) is one of the most widely used biocatalysts in organic synthesis. The traditional method for purification of CalB is a multi-step, high cost and low recovery procedure. Biomimetic affinity purification had high efficiency purification. We selected 298 ligand columns from a 700-member library of synthetic ligands to screen Pichia pastoris protein extract. Of the 298, three columns (named as A9-14, A9-10, and A11-33) had one-step purification effect, and A9-14 of these affinity ligands, had both high purification and recovery. The one-step recovery of CalB reached 73% and the purification reached 91% upon purification. The active groups of A9-14 were cyclohexylamine and propenylamine. Furthermore, both A9-14 and A9-10 had the same R1 active group of cyclohexylamine which might act the main binding role for CalB. The synthetic ligand A9-14 had a binding capacity of 0.4 mg/mL and had no negative effects on its hydrolytic activity. Unlike a natural affinity ligand, this synthetic ligand is highly stable to resist 1M NaOH, and thus has great potential for industrial scale production of CalB.
authors.affiliation: Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Yao
authors.forename: Tian
authors.initials: T
authors.lastname: Zhang
authors.forename: Hongwei
authors.initials: H
authors.lastname: Xue
authors.forename: Kexuan
authors.initials: K
authors.lastname: Tang
authors.forename: Rongxiu
authors.initials: R
authors.lastname: Li
mesh_terms: D040761:Biomimetic Materials / Q000737:chemistry* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D004795:Enzyme Stability; D005656:Fungal Proteins; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010843:Pichia / Q000737:chemistry; D011485:Protein Binding; D054852:Small Molecule Libraries; D012972:Sodium Hydroxide
chemical_list: D005656:Fungal Proteins; D054852:Small Molecule Libraries; D012972:Sodium Hydroxide; D008049:Lipase; C431349:lipase B, Candida antarctica