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Open AccessArticle

Developing a Novel Enzyme Immobilization Process by Activation of Epoxy Carriers with Glucosamine for Pharmaceutical and Food Applications

1
Department of Food, Environmental and Nutritional Sciences, University of Milan, Via Mangiagalli 25, I-20133 Milano, Italy
2
Department of Agriculture and Forestry Science (DAFNE), University of Tuscia, Via S. Camillo de Lellis snc, 01100 Viterbo, Italy
3
Department of Drug Sciences, University of Pavia, Viale Taramelli 12, I-27100 Pavia, Italy
4
Department of Biology and Biotechnology Lazzaro Spallanzani, University of Pavia, Via Ferrata 9, I-27100 Pavia, Italy
*
Author to whom correspondence should be addressed.
Catalysts 2019, 9(10), 843; https://doi.org/10.3390/catal9100843
Received: 3 September 2019 / Revised: 6 October 2019 / Accepted: 8 October 2019 / Published: 12 October 2019
(This article belongs to the Special Issue Immobilization of Enzymes)
In this paper, we describe the development of an efficient enzyme immobilization procedure based on the activation of epoxy carriers with glucosamine. This approach aims at both creating a hydrophilic microenvironment surrounding the biocatalyst and introducing a spacer bearing an aldehyde group for covalent attachment. First, the immobilization study was carried out using penicillin G acylase (PGA) from Escherichia coli as a model enzyme. PGA immobilized on glucosamine activated supports has been compared with enzyme derivatives obtained by direct immobilization on the same non-modified carriers, in the synthesis of different 3′-functionalized cephalosporins. The derivatives prepared by immobilization of PGA on the glucosamine-carriers performed better than those prepared using the unmodified carriers (i.e., 90% versus 79% cefazolin conversion). The same immobilization method has been then applied to the immobilization of two other hydrolases (neutral protease from Bacillus subtilis, PN, and bromelain from pineapple stem, BR) and one transferase (γ-glutamyl transpeptidase from Bacillus subtilis, GGT). Immobilized PN and BR have been exploited in the synthesis of modified nucleosides and in a bench-scale packed-bed reactor for the protein stabilization of a Sauvignon blanc wine, respectively. In addition, in these cases, the new enzyme derivatives provided improved results compared to those previously described. View Full-Text
Keywords: enzyme immobilization; glucosamine; epoxy carrier; penicillin G acylase; protease N; bromelain; γ-glutamyl transpeptidase enzyme immobilization; glucosamine; epoxy carrier; penicillin G acylase; protease N; bromelain; γ-glutamyl transpeptidase
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Serra, I.; Benucci, I.; Robescu, M.S.; Lombardelli, C.; Esti, M.; Calvio, C.; Pregnolato, M.; Terreni, M.; Bavaro, T. Developing a Novel Enzyme Immobilization Process by Activation of Epoxy Carriers with Glucosamine for Pharmaceutical and Food Applications. Catalysts 2019, 9, 843.

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