title: Recovery of recombinant cutinase using detergent foam.
abstract: Foam generated by vigorous stirring of a nonionic detergent, Triton X-114, was used for the recovery of recombinant cutinase expressed by Saccharomyces cerevisiae. The enzyme with a hydrophobic fusion tag, (Trp-Pro)(4), was recovered with a higher yield as compared to the wild-type cutinase, indicating the involvement of hydrophobic interactions in protein isolation with the foam. The influence of various factors including volume, dilution, pH, different additives, and cell concentration in the medium on enzyme recovery was investigated. Interaction of the enzyme with detergent was monitored using fluorescence spectroscopy. No significant changes in protein conformation after the isolation procedure were observed using circular dichroism.
authors.affiliation: Department of Biotechnology, Center for Chemistry & Chemical Engineering, Lund University, P.O. Box 124, S-221 00 Lund, Sweden.
authors.forename: Sheryl
authors.initials: S
authors.lastname: Fernandes
authors.forename: Bo
authors.initials: B
authors.lastname: Mattiasson
authors.forename: Rajni
authors.initials: R
authors.lastname: Hatti-Kaul
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D003902:Detergents / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000737:chemistry; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D003902:Detergents; D011993:Recombinant Fusion Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D002265:Carboxylic Ester Hydrolases; C019952:cutinase