Immobilization and stabilization of xylanase by multipoint covalent attachment on agarose and on chitosan supports.
Xylanases have important applications in industry. Immobilization and stabilization of enzymes may allow their reuse in many cycles of the reaction, decreasing the process costs. This work proposes the use of a rational approach to obtain immobilized commercial xylanase biocatalysts with optimized features. Xylanase NS50014 from Novozymes was characterized and immobilized on glyoxyl-agarose, agarose-glutaraldehyde, and agarose-amino-epoxy support and on differently activated chitosan supports: glutaraldehyde-chitosan, glyoxyl-chitosan, and epoxy-chitosan. Two different chitosan matrices were tested. The best chitosan derivative was epoxy-chitosan-xylanase, which presented 100% of immobilization yield and 64% of recovered activity. No significant increase on the thermal stability was observed for all the chitosan-enzyme derivatives. Immobilization on glyoxyl-agarose showed low yield immobilization and stabilization degrees of the obtained derivative. The low concentration of lysine groups in the enzyme molecule could explain these poor results. The protein was then chemically modified with ethylenediamine and immobilized on glyoxyl-agarose. The new enzyme derivatives were 40-fold more stable than the soluble, aminated, and dialyzed enzyme (70 degrees C, pH 7), with 100% of immobilization yield. Therefore, the increase of the number of amine groups in the enzyme surface was confirmed to be a good strategy to improve the properties of immobilized xylanase.
Manrich
Anny
A
Chemical Engineering Department, Universidade Federal de São Carlos, Rodovia Washington Luís, Km 235, São Carlos, SP, Brazil.
Komesu
Andrea
A
Adriano
Wellington Sabino
WS
Tardioli
Paulo Waldir
PW
Giordano
Raquel Lima Camargo
RL
D000818:Animals; D048271:Chitosan / Q000737:chemistry*; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry / Q000378:metabolism; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005782:Gels / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D012685:Sepharose / Q000737:chemistry*; D013696:Temperature
D004800:Enzymes, Immobilized; D005782:Gels; D012685:Sepharose; D048271:Chitosan; D043364:Endo-1,4-beta Xylanases