title: Micro-scale procedure for enzyme immobilization screening and operational stability assays.
abstract: OBJECTIVE
  A simple and inexpensive methodology, based on the use of micro-centrifuge filter tubes, is proposed for establishing the best enzyme immobilization conditions.
  
  
  RESULTS
  The immobilized biocatalyst is located inside the filter holder during the whole protocol, thus facilitating the incubations, filtrations and washings. This procedure minimizes the amount of enzyme and solid carrier needed, and allows exploring different immobilization parameters (pH, buffer concentration, enzyme/carrier ratio, incubation time, etc.) in a fast manner. The handling of immobilized enzymes using micro-centrifuge filter tubes can also be applied to assess the apparent activity of the biocatalysts, as well as their reuse in successive batch reaction cycles. The usefulness of the proposed methodology is shown by the determination of the optimum pH for the immobilization of an inulinase (Fructozyme L) on two anion-exchange polymethacrylate resins (Sepabeads EC-EA and Sepabeads EC-HA).
  
  
  CONCLUSIONS
  The micro-scale procedure described here will help to overcome the lack of guidelines that usually govern the selection of an immobilization method, thus favouring the development of stable and robust immobilized enzymes that can withstand harsh operating conditions in industry.
authors: Instituto de Catálisis y Petroleoquímica, CSIC, Cantoblanco, Marie Curie 2, 28049, Madrid, Spain.
authors: Lucia
authors: L
authors: Fernandez-Arrojo
authors: Paloma
authors: P
authors: Santos-Moriano
authors: Barbara
authors: B
authors: Rodriguez-Colinas
authors: Antonio O
authors: AO
authors: Ballesteros
authors: Francisco J
authors: FJ
authors: Plou
mesh_terms: D004800:Enzymes, Immobilized / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008403:Mass Screening / Q000379:methods*; D011109:Polymethacrylic Acids; D011485:Protein Binding
chemical_list: D004800:Enzymes, Immobilized; D011109:Polymethacrylic Acids; C030613:polymethacrylic acid; D006026:Glycoside Hydrolases; C023086:inulinase