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

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: Process Biochemistry; v. 95, p. 251-259, AUG 2020.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 04/14593-4 - Síntese de biodiesel por interesterificação enzimática de óleos vegetais empregando lipase estabilizada pela técnica multipontual
• Beneficiário: ADRIANO AGUIAR MENDES
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 06/53596-4 - Imobilizacao/estabilizacao de d hidantoinase para producao de n carbamoil d fenilglicina.
• Beneficiário: Aline Aparecida Becaro
• Modalidade de apoio: Bolsas no Brasil - Mestrado