Reversed micellar extraction of charged fusion proteins.
We have investigated the use of charged fusion tails with the enzyme glucoamylase in reversed micellar extraction. The addition of the charged tails increased the fraction of enzymatically active protein recovered at a given pH, with the tails containing the largest number of charges being recovered at the highest level. The series of mutations also allows for investigation of the charge-dependent behavior of reversed micellar extraction. However, in this case, the change in protein charge via fusions had a lesser impact than did the change in charge via a pH change. The difference may be due to the difficulty of partitioning the hydrodynamically larger fusion protein.
Forney
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CE
Department of Chemical Engineering, Iowa State University, Ames 50011-2230.
Glatz
C E
CE
D000426:Alcohol Dehydrogenase / Q000302:isolation & purification; D000595:Amino Acid Sequence; D001230:Aspergillus / Q000201:enzymology / Q000235:genetics; D004563:Electrochemistry; D005087:Glucan 1,4-alpha-Glucosidase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008823:Micelles; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D012710:Serum Albumin, Bovine / Q000302:isolation & purification
D008823:Micelles; D011993:Recombinant Fusion Proteins; D012710:Serum Albumin, Bovine; D000426:Alcohol Dehydrogenase; D005087:Glucan 1,4-alpha-Glucosidase