Immobilization of a recombinant Escherichia coli producing a thermostable alpha-L-rhamnosidase: Creation of a bioreactor for hydrolyses of naringin

Research output: Contribution to journalArticle

Abstract

An U-L-rhamnosidase (E.C. 3.2.1.40) from a newly discovered thermophilic bacterium was expressed in Escherichia coli BL21 DE3 pRIL cells. The cells were immobilized in Ca2+-alginate beads. The temperature of 50 degrees C used in reactions, appeared to be sufficient for making the mesophilic strain porous enough for the substrate to access the cloned thermostable enzyme. Pretreatment of cells with heat or lysozyme prior to bead formation did not improve the results. The best cell concentration (w/w) for bead preparation was found to be 0.0 192 g ml(-1) and stability of beads increased if CaCl2 concentration in buffers and substrate was kept at 50 mM. In a 60 min assay, the optimal pH of the entrapped cells was found to be 7.8 and the optimal temperature 60 degrees C. By packing the beads in a column, a bioreactor for production Of L-rhamnose from naringin was created. Full degradation of 7.9 mM naringin could be reached by running the reactor at 1 ml min(-1) at 50 degrees C. The optimal running temperature of the reactor was found to be 50 degrees C and the reactor was fully stable over 3 days at that temperature. On the fourth day, substrate degradation capacity had decreased by 10-15%. (c) 2006 Elsevier Inc. All rights reserved.

Details

Authors
  • Hakon Birgisson
  • Jon Oskar Wheat
  • Gudmundur O. Hreggvidsson
  • Jakob K. Kristjansson
  • Bo Mattiasson
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Chemical Sciences

Keywords

  • recombinant alpha-L-rhamnosidase, immobilization, alginate, thermophilic, bioreactor
Original languageEnglish
Pages (from-to)1181-1187
JournalEnzyme and Microbial Technology
Volume40
Issue number5
Publication statusPublished - 2007
Publication categoryResearch
Peer-reviewedYes

Bibliographic note

The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Center for Chemistry and Chemical Engineering (011001000), Biotechnology (LTH) (011001037)