[自然科学]生物分离工程-双语版 3.pptVIP

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[自然科学]生物分离工程-双语版 3.ppt

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[自然科学]生物分离工程-双语版 3

Chapter 31st High resolution fractionation processes I Introduction (1) appropriate sequence (2) utilization (therapeutic, diagnostic) (3) operation scale (4) role of final fractionation A impurities B salts C pathogens D endotoxins E contaminating proteins (5) low volume and expensive procedures (6) special attention A amount of product denaturation B quality II Chromatography procedures A gel filtration chromatography B Hydroxylapatite chromatography C Hydrophobic interaction chromatography D Affinity chromatography E Ion-Exchange Chromatography A gel filtration chromatography (1) mechanism A molecular size B mobile solvent phase C stationary phase (2) Rigidity of beads in large scale operation (3) traditional gel filtration chromatography media A Sephadex B Superose C Sephacryl D Trisacryl E Biogel P F Cellulofine G Fractogel (4) low product volume suitable for gel filtration (5) activity loss A final fractionation B previous bioseparation steps Exmaple 3.1 carboxylesterase purification (1) Material ：Bacillus stearothermophilus噬热脂杆菌 (2) Step utilized A centrifugation B DEAE-Sephacel ion-exchange chromatography C gel filtration chromatography (3) Result analysis A single low-mobility band (PAGE) B Two bands of higher mobility (after precipitation and anion exchange） C just single band of high mobility (further gel filtration) D gel filtration and SDSindicate esterase is monomeric protein with MW 40000 Exmaple 3.1 carboxylesterase purification (4) Reason A active monomer associating multiple forms B multiple form have different chromatography and electrophoretic properties C reaction equilibrium affected by pH, salt concentration, enzyme dilution D monomers or multimeric enzyme forms affect activity and stability Difference between c