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Recovery of Recombinant Cutinase Using Detergent Foam
Author(s) -
Fernandes Sheryl,
Mattiasson Bo,
HattiKaul Rajni
Publication year - 2002
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp010161m
Subject(s) - cutinase , chemistry , chromatography , circular dichroism , recombinant dna , dilution , enzyme , saccharomyces cerevisiae , biochemistry , yeast , gene , thermodynamics , physics
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.