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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Immobilization and stabilization of D-hydantoinase from Vigna angularis and its use in the production of N-carbamoyl-D-phenylglycine. Improvement of the reaction yield by allowing chemical racemization of the substrate

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Author(s):
Aparecida Becaro, Aline [1] ; Aguiar Mendes, Adriano [2] ; Sabino Adriano, Wellington [3] ; Antunes Lopes, Laiane [4] ; Lourenco Vanzolini, Kenia [5] ; Fernandez-Lafuente, Roberto [6, 7] ; Waldir Tardioli, Paulo [4] ; Bezerra Cass, Quezia [5] ; Camargo Giordano, Raquel de Lima [4]
Total Authors: 9
Affiliation:
[1] Univ Fed Sao Carlos, Postgrad Program Biotechnol PPGBiotec, Rod Washington Luis, Km 235, BR-1365905 Sao Paulo, SP - Brazil
[2] UNIFAL, Inst Chem, BR-37130001 Alfenas, MG - Brazil
[3] UFCG, Educ & Hlth Ctr, Cuite, PB - Brazil
[4] Univ Fed Sao Carlos, DEQ, Postgrad Program Chem Engn PPGEQ, Rod Washington Luis, Km 235, BR-13565905 Sao Carlos, SP - Brazil
[5] Univ Fed Sao Carlos, Dept Chem, Rod Washington Luis, Km 235, BR-13565905 Sao Carlos, SP - Brazil
[6] CSIC, ICP, Dept Biocatalisis, Campus UAM CSIC, Madrid 28049, Spain.Aparecida Becaro, Aline, Univ Fed Sao Carlos, Postgrad Program Biotechnol PPGBiotec, Rod Washington Luis, Km 235, BR-1365905 Sao Paulo, SP - Brazil
[7] CSIC, ICP, Dept Biocatalisis, Campus UAM CSIC, Madrid 28049 - Spain
Total Affiliations: 7
Document type: Journal article
Source: Process Biochemistry; v. 95, p. 251-259, AUG 2020.
Web of Science Citations: 0
Abstract

This work reports the immobilization of a multimeric D-hydantoinase (DHTase) from Vigna angularis (E.C. 3.5.2.2.) on agarose beads activated with glyoxyl groups aiming to improve its stability via multipoint covalent attachment. The final reduction with sodium borohydride resulted in a drop in enzyme activity that could be decreased by adding Zn2+ or Mg2+. The optimal preparation with high activity (58 % recovered activity) and stability (around 86-fold more stable than the free enzyme) was obtained by DHTase immobilization on glyoxyl agarose for 24 h at 25 degrees C and pH 10.05, and a borohydride reduction step in the presence of 10 mM Zn2+ (DHTase-Glx). The enzyme was almost fully immobilized on glyoxyl agarose (19.8 mg/g of support) when offering 20 mg/g. This immobilized biocatalyst was used to catalyze the hydrolysis of D,L-phenylhydantoin under substrate racemization conditions, which produced 99 % of N-carbamoyl-D-phenylglycine after 9 h reaction. (AU)