Evaluation of microbeads of calcium alginate as a fluidized bed medium for affinity chromatography of Aspergillus niger pectinase

• Published in: Biotechnology progress Vol. 20; no. 5; pp. 1490 - 1495
• Main Authors: Roy, I, Jain, S, Teotia, S, Gupta, M.N
• Format: Journal Article
• Language: English
• Published: USA American Chemical Society 2004; American Institute of Chemical Engineers
• Subjects: Affinity chromatography; Alginates - chemistry; Aspergillus niger; Aspergillus niger - enzymology; Beads; Biological and medical sciences; Biotechnology; calcium alginate; Chromatography, Affinity - methods; Coated Materials, Biocompatible - chemistry; Coated Materials, Biocompatible - isolation & purification; Enzymes, Immobilized - biosynthesis; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - isolation & purification; Fluidized beds; Fundamental and applied biological sciences. Psychology; Glucuronic Acid - chemistry; Hexuronic Acids - chemistry; Materials Testing; Microfluidics - instrumentation; Microfluidics - methods; Microspheres; Polygalacturonase; Polygalacturonase - biosynthesis; Polygalacturonase - isolation & purification; Q1; Q2; Surface Properties; Calcium; Enzyme; Alginates; Polygalacturonase; Fungi; Aspergillus niger; Affinity chromatography; Glycosidases; Hydrolases; Fungi Imperfecti; O-Glycosidases; Thallophyta; Fluidized bed
• ISSN: 8756-7938; 1520-6033
• DOI: 10.1021/bp0343864

Calcium alginate microbeads (212–425 μm) were prepared by spraying 2% (w/v) alginate solution into 1 M CaCl2 solution. The fluidization behavior of these beads was studied, and the bed expansion index and terminal velocity were found to be 4.3 and 1808 cm h−1, respectively. Residence time distribution curves showed that the dispersion of the protein was much less with these microbeads than with conventionally prepared calcium alginate macrobeads when both kinds of beads were used for chromatography in a fluidized bed format. The fluidized bed of these beads was used for the purification of pectinase from a commercial preparation. The media performed well even with diluted feedstock; 90% activity recovery with 211‐fold purification was observed.