Evaluation of microbeads of calcium alginate as a fluidized bed medium for affinity chromatography of Aspergillus niger pectinase
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 distributi...
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| Published in: | Biotechnology progress Vol. 20; no. 5; pp. 1490 - 1495 |
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| Language: | English |
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| Abstract | 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. |
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| AbstractList | Calcium alginate microbeads (212-425 microm) 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. Calcium alginate microbeads (212-425 mu m) were prepared by spraying 2% (w/v) alginate solution into 1 M CaCl sub(2) 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 super(-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. 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. |
| Author | Gupta, Munishwar Nath Teotia, Sunita Roy, Ipsita Jain, Sulakshana |
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| Keywords | Calcium Enzyme Alginates Polygalacturonase Fungi Aspergillus niger Affinity chromatography Glycosidases Hydrolases Fungi Imperfecti O-Glycosidases Thallophyta Fluidized bed |
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| References_xml | – volume: 8 start-page: 235 year: 2001 end-page: 239 article-title: Separation of phospholipase D from peanut on a fluidized bed of cross‐linked alginate beads publication-title: Biochem. Eng. J – volume: 68 start-page: 175 year: 2002 end-page: 204 article-title: Downstream processing of enzymes/proteins publication-title: Proc. Indian Natl. Sci. Acad., Part B – volume: 15 start-page: 230 year: 1997 end-page: 235 article-title: Expanded bed adsorption in industrial bioprocessing: Recent developments publication-title: Trends Biotechnol. – volume: 865 start-page: 129 year: 1999 end-page: 144 article-title: Expandedbed chromatography in primary protein purification publication-title: J. Chromatogr. A – start-page: 194 year: 1979 – volume: 18 start-page: 111 year: 2000 end-page: 114 article-title: Purification of wheat germ amylase by precipitation publication-title: Protein Expression Purif. – volume: 28 start-page: 190 year: 2003 end-page: 195 article-title: Macroaffinity ligand facilitated three phase partitioning (MLFTPP) for purification of glucoamylase and pullulanase using alginate publication-title: Protein Expression Purif. – volume: 950 start-page: 131 year: 2002 end-page: 137 article-title: Purification of a bacterial pullulanase on a fluidized bed of calcium alginate beads publication-title: J. Chromatogr. A – volume: 1 start-page: 199 year: 1998 end-page: 226 – volume: 27 start-page: 53 year: 2000 end-page: 65 article-title: Exploiting unusual affinity of usual polysaccharides for bioseparation of enzymes on fluidized beds publication-title: Enzyme Microb. Technol. – volume: 28 start-page: 792 year: 2001 end-page: 795 article-title: An efficient purification process for sweet potato β‐amylase by affinity precipitation with alginate publication-title: Enzyme Microb. Technol. – volume: 32 start-page: 35 year: 1954 end-page: 53 article-title: Sedimentation and fluidization. Part I publication-title: Trans. 1nst. Chem. Eng. – start-page: 363 year: 1993 end-page: 399 – year: 1986 – volume: 12 start-page: 266 year: 1990 end-page: 271 article-title: Strain and process for production of polygalacturonase publication-title: Enzyme Microb. Technol. – volume: 18 start-page: 321 year: 1993 end-page: 324 article-title: Purification of endopolygalacturonase by affinity precipitation using alginate publication-title: Biotechnol. Appl. Biochem. – volume: 40 start-page: B7 year: 1989 end-page: B19 article-title: Isolation and purification of endo‐polygalacturonase by affinity chromatography in a fluidized bed reactor publication-title: Chem. Eng. J. – volume: 72 start-page: 248 year: 1976 end-page: 254 article-title: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein‐dye binding publication-title: Anal. Biochem. – volume: 11 start-page: 211 year: 1998 end-page: 216 article-title: Affinity precipitation of proteins: design criteria for an efficient polymer publication-title: J. Mol. Recognit. – year: 1969 – volume: 28 start-page: 499 year: 2001 end-page: 509 article-title: Combined effect of operational variables and enzyme activity on aqueous enzymatic extraction of oil and protein from soybean publication-title: Enzyme Microb. Technol. – year: 1972 – volume: 31 start-page: 426 year: 1959 end-page: 428 article-title: Useof dinitrosalicylic acid reagent for determination of reducing sugar publication-title: Anal. Chem. – volume: 12 start-page: 296 year: 1994 end-page: 303 article-title: Purificationof proteins by adsorption chromatography in expanded beds publication-title: Trends Biotechnol. – volume: 35 start-page: 161 year: 2001 end-page: 165 article-title: Alginate as a macroaffinity ligand and an additive for enhanced activity and thermostability of lipases publication-title: Biotechnol. Appl. Biochem. – volume: 995 start-page: 127 year: 2003 end-page: 134 article-title: Separation of enzyme by sequential macroaffinity ligand facilitated three phase partitioning publication-title: J. Chromatogr. A |
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| Snippet | 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... Calcium alginate microbeads (212-425 microm) were prepared by spraying 2% (w/v) alginate solution into 1 M CaCl2 solution. The fluidization behavior of these... Calcium alginate microbeads (212-425 mu m) were prepared by spraying 2% (w/v) alginate solution into 1 M CaCl sub(2) solution. The fluidization behavior of... |
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| SubjectTerms | 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 |
| Title | Evaluation of microbeads of calcium alginate as a fluidized bed medium for affinity chromatography of Aspergillus niger pectinase |
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