A comparative analysis of catalytic activity and stability of microbial transglutaminase in controlled denaturing conditions(42 visite) Caporale A, Monti A, Selis F, Sandomenico A, Tonon G, Ruvo M, Doti N
J Biotechnol (ISSN: 0168-1656linking), 2019 Aug 20; 302: 48-57.
Tipo di articolo: Journal Article
Impact factor: 3.163, Impact factor a 5 anni: 0
Url: Non disponibile.
Parole chiave: Biocatalysis, Enzyme Stability, Protein Denaturation, Spectrometry, Fluorescence, Substrate Specificity, Transglutaminases, Chemistry, Metabolism, Bioconjugation, Chemical Denaturation, Circular Dichroism (cd), Fluorescence Spectroscopy, Microbial Transglutaminase (mtg)
*** IBB - CNR *** IBB-CNR, Via Mezzocannone 16, 80134, Napoli, Italy., IBB-CNR, Via Mezzocannone 16, 80134, Napoli, Italy; Universita degli studi della Campania "Luigi Vanvitelli", Via Vivaldi n. 43 - 81100 Caserta, Caserta, Italy., BIOVIIIx, via Brin, 59, 80142, Napoli, Italy., IBB-CNR, Via Mezzocannone 16, 80134, Napoli, Italy. Electronic address: firstname.lastname@example.org., IBB-CNR, Via Mezzocannone 16, 80134, Napoli, Italy. Electronic address: email@example.com.,
Microbial transglutaminases (MTGs) catalyzes the formation of Gln-Lys isopeptide bonds and are widely used for the cross-linking of proteins and peptides in food and in biotechnological applications for bioconjugation reactions. In view of its practical utility, a comparative study of the catalytic activity and stability of the enzyme in a wide range of denaturing conditions has been performed through Circular Dichroism (CD), fluorescence and activity assays performed with model substrates. In agreement with previous results, we show that MTG has a significant structural and functional tolerance to pH changes, whereas the enzyme stability and activity decrease in presence of increasing amounts of denaturing agents, such as urea and guanidinium chloride (GdnHCl). Noteworthy, the activity of MTG in denaturing conditions differs markedly from that in pseudo-physiological settings, shifting unexpectedly toward higher substrate specificity. Also, the use of controlled amounts of denaturing agents (1.0-1.5M urea) largely improves yields and purity of the final products of 10-15% and 25-30%, respectively. These findings widen the range of applicability of the MTG-mediated biocatalysis for industrial and biotechnological purposes.
378 Records (370 escludendo Abstract e Conferenze). Impact factor totale: 1486.231 (1463.34 escludendo Abstract e Conferenze). Impact factor a 5 anni totale: 1527.634 (1503.207 escludendo Abstract e Conferenze).