title: M(III)-facilitated recovery and concentration of enzymes from mesophilic and thermophilic organisms.
abstract: Six enzymes isolated from organisms of widely differing thermal growth optima were flocculated from solution at constant pH by addition of Fe(III) solution. In all cases the enzyme concentration was 1 g.l-1 or less. Flocculation profiles were generated for each enzyme over a range of Fe(III) levels. The concentrated enzymes were recovered from the Fe(III)/protein complex by solubilisation with citrate and dithionite followed by precipitation with ammonium sulphate. In all cases approximately 70-80% enzyme recovery was achieved. Enzyme thermal stability did not appear to be important and protein concentration had no effect on the efficiency of enzyme recovery over the range of 0.01-1 g.l-1. Approximately 30 mmol Fe(III)/l of enzyme solution facilitated optimal enzyme recovery for all solutions studied. For protein concentrations up to 1 g.l-1 a 100-fold enzyme concentration factor can be expected.
authors.affiliation: Chemistry Department, University of Waikato, Hamilton, New Zealand.
authors.forename: T N
authors.initials: TN
authors.lastname: Collingwood
authors.forename: M
authors.initials: M
authors.lastname: Shanmugam
authors.forename: R M
authors.initials: RM
authors.lastname: Daniel
authors.forename: A G
authors.initials: AG
authors.lastname: Langdon
mesh_terms: D000535:Aluminum; D001407:Bacillus / Q000201:enzymology*; D001412:Bacillus subtilis / Q000201:enzymology; D002480:Cellulase; D004795:Enzyme Stability; D005427:Flocculation; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D007501:Iron; D003904:Mitosporic Fungi / Q000201:enzymology*; D010447:Peptide Hydrolases / Q000302:isolation & purification*; D013816:Thermodynamics; D013824:Thermus / Q000201:enzymology*
chemical_list: D000535:Aluminum; D007501:Iron; D006026:Glycoside Hydrolases; D002480:Cellulase; C019826:carboxymethylcellulase; D010447:Peptide Hydrolases