From Batch to Continuous Flow Bioprocessing : Use of an Immobilized J-Glutamyl-Transferase from B. subtilis for the Synthesis of Biologically Active Peptide Derivatives
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Data di Pubblicazione:
2021
Citazione:
From Batch to Continuous Flow Bioprocessing : Use of an Immobilized J-Glutamyl-Transferase from B. subtilis for the Synthesis of Biologically Active Peptide Derivatives / M.S. Robescu, F. Annunziata, V. Somma, C. Calvio, C.F. Morelli, A. Pinto, G. Speranza, L. Tamborini, D. Ubiali, T. Bavaro. ((Intervento presentato al 6. convegno BNCM: International Conference on Biocatalysis in Non-Conventional Media tenutosi a online-Milano nel 2021.
Abstract:
J-Glutamyl peptides (J-E-peptides) are raising increasing interest due to their
numerous biological properties (e.g., taste enhancement, antioxidant, anticancer,
anti-inflammatory and lipid-lowering activity).
J-Glutamyl transferases (GGTs, E.C.
2.3.2.2) are naturally involved in the transfer of J-E-moieties from different J-Edonors to both amino acids and peptides, thus producing J-E-derivatives (Scheme).
Scheme. GGT-Catalyzed synthesis of J-glutamyl-derivatives (1-4)
The GGT from Bacillus subtilis (BsGGT) was covalently immobilized on glyoxylagarose (GLX-AG) resulting in high protein immobilization yield and activity recovery
(>95%). BsGGT-GLX-AG was shown to be a robust biocatalyst in the batch
synthesis of biologically active J-E-peptides (1-4) in fully aqueous medium (isolated
yields: 19-40%, 5-10 g L-1). BsGGT-GLX-AG was recovered and re-used for five
consecutive reaction cycles; the biocatalyst retained 90% activity under storage
conditions (10 months, 4 °C).
Based on these results, we moved from batch to continuous flow biotransformation
with the aim to control side-reactions (i.e., 5-8) and to achieve process
intensification. Under the optimized conditions to date, the productivity of the model
compound J-E-S-allyl-cysteine (2) was increased by a two-fold factor (7.8 Pmol min-1
g-1) and the impurity profile was improved as well. Further optimization under
continuous flow conditions is currently in progress.
Tipologia IRIS:
14 - Intervento a convegno non pubblicato
Elenco autori:
M.S. Robescu, F. Annunziata, V. Somma, C. Calvio, C.F. Morelli, A. Pinto, G. Speranza, L. Tamborini, D. Ubiali, T. Bavaro
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