Immobilization of the glucose isomerase from Caldicoprobacter algeriensis on Sepabeads EC-HA and its efficient application in continuous High Fructose Syrup production using packed bed reactor

Sawssan Neifar; Fadia V Cervantes; Amel Bouanane-Darenfed; Hajer BenHlima; Antonio O Ballesteros; Francisco J Plou; Samir Bejar

doi:10.1016/j.foodchem.2019.125710

Immobilization of the glucose isomerase from Caldicoprobacter algeriensis on Sepabeads EC-HA and its efficient application in continuous High Fructose Syrup production using packed bed reactor

Food Chem. 2020 Mar 30:309:125710. doi: 10.1016/j.foodchem.2019.125710. Epub 2019 Oct 19.

Authors

Sawssan Neifar¹, Fadia V Cervantes², Amel Bouanane-Darenfed³, Hajer BenHlima⁴, Antonio O Ballesteros⁵, Francisco J Plou⁶, Samir Bejar⁷

Affiliations

¹ Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.
² Institute of Catalysis and Petrochemistry, CSIC, Marie Curie 2, 28049 Madrid, Spain.
³ Laboratory of Cellular and Molecular Biology (LCMB), Microbiology Team, Faculty of Biological Sciences, University of Sciences and Technology of Houari Boumediene (USTHB), PO Box 32, El Alia, Bab Ezzouar 16111, Algeria.
⁴ Unit of Algae Biotechnology, Biological Engineering Department, National School of Engineers of Sfax, University of Sfax, Sfax 3038, Tunisia.
⁵ Institute of Catalysis and Petrochemistry, CSIC, Marie Curie 2, 28049 Madrid, Spain. Electronic address: a.ballesteros@icp.csic.es.
⁶ Institute of Catalysis and Petrochemistry, CSIC, Marie Curie 2, 28049 Madrid, Spain. Electronic address: fplou@icp.csic.es.
⁷ Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia. Electronic address: samir.bejar@cbs.rnrt.tn.

PMID: 31704076
DOI: 10.1016/j.foodchem.2019.125710

Abstract

The glucose isomerase GICA from Caldicoprobacter algeriensis was immobilized by ionic adsorption on polymethacrylate carriers (Sepabeads EC-EA and EC-HA) or covalent attachment to glyoxal agarose. The Sepabeads EC-HA yielded the highest recovery of activity (89%). The optimum temperature and pH of immobilized GICA were 90 °C and 7.0, respectively, similar to the corresponding values of free enzyme. Nevertheless, the adsorbed enzyme displayed higher relative activity at acidic pH, greater thermostability, and better storage stability, compared to the free form. Moreover, the immobilized enzyme showed an excellent operational stability, in 15 successive 3 h reaction cycles at 85 °C under a batch reactor, preserving 83% of its initial activity. Interestingly, a continuous process for High Fructose Syrup (HFS) production was established with the adsorbed GICA using a packed bed reactor during eleven days at 70 °C. HPAEC-PAD analysis showed a maximum bioconversion rate of 49% after 48 h of operation.

Keywords: Batch reactor; Enzyme immobilization; Glucose isomerase; HFS; Packed bed reactor; Sepabeads; Sodium borohydride (PubChem CID: 4311764).

MeSH terms

Aldose-Ketose Isomerases / chemistry
Aldose-Ketose Isomerases / metabolism*
Batch Cell Culture Techniques / methods*
Clostridiales / enzymology*
Enzyme Stability
Enzymes, Immobilized / chemistry
Enzymes, Immobilized / metabolism
Fructose / metabolism*
Hydrogen-Ion Concentration
Sepharose / chemistry
Temperature

Substances

Enzymes, Immobilized
Fructose
Sepharose
Aldose-Ketose Isomerases
xylose isomerase

Supplementary concepts

Caldicoprobacter algeriensis