Introducció a les Xarxes Telemàtiques: problemes amb solució

Col·lecció de problemes resolts d'IXT (Introducció a les Xarxes Telemàtiques) amb solució, 1er curs dels graus de l'ETSETB.2022/20232n quadrimestr

Introducció a les Xarxes Telemàtiques: problemes amb solució

Col·lecció de problemes resolts d'IXT (Introducció a les Xarxes Telemàtiques) amb solució, 1er curs dels graus de l'ETSETB2022/20232n quadrimestr

Recombinant production of serine hydroxymethyl transferase from Streptococcus thermophilus and its preliminary evaluation as a biocatalyst

9 pages, 3 figures, 3 tables.-- PMID: 15726349 [PubMed].-- Printed version published Sep 2005.The glyA gene encoding a serine hydroxymethyl transferase (SHMT) with threonine aldolase activity was isolated from Streptococcus thermophilus YKA-184 chromosomal DNA. This aldolase is a pyridoxal 5′-phosphate-dependent enzyme that stereospecifically catalyzes the interconversion of l-threonine to glycine and acetaldehyde. The enzyme was overexpressed in Escherichia coli M15 as a recombinant protein of 45 kDa with a His6-tag at its N-terminus. The recombinant enzyme was purified to homogeneity by a single chromatographic step using Ni-nitrilotriacetic acid affinity, obtaining a high activity-recovery yield (83%). Lyophilized and precipitated enzymes were stable at least for 10 weeks when stored at −20°C and 4°C. It was observed that the K m for l-allo-threonine was 38-fold higher than that for l-threonine, suggesting this enzyme can be classified as a specific l-allo-threonine aldolase. The optimum pH range of threonine aldolase activity for the recombinant SHMT was pH 6–7. When tested for aldol addition reactions with non-natural aldehydes, such as benzyloxyacetaldehyde and (R)-N-Cbz-alaninal, two possible β-hydroxy-α-amino acid diastereoisomers were produced, but with moderate stereospecificity. The enzyme showed potential as a biocatalyst for the stereoselective synthesis of β-hydroxy-α-amino acids.Financial support from CICYT (project PPQ 2002-04625-C02-01).Peer reviewe

Chemoenzymatic Synthesis and Inhibitory Activities of Hyacinthacines A1 and A2 Stereoisomers.

6 páginas,1 figura, 2 esquemas, 2 tablas.A novel straightforward chemoenzymatic procedure for the synthesis of hyacinthacine stereoisomers based on the aldol addition of dihydroxyacetone phosphate (DHAP) to N-Cbz-prolinal under catalysis by l-rhamnulose 1-phosphate aldolase from E. coli is presented. The synthesis is complemented by a simple and effective purification protocol consisting of ion-exchange chromatography on CM-sepharose. As examples, ( )-hyacinthacine A2 [the enantiomer of (+)-hyacinthacine A2], 7-deoxy- 2-epialexine (the enantiomer of 3-epihyacinthacine A2), ent-7-deoxyalexine (the enantiomer of 7-deoxyalexine) and 2-epihyacinthacine A2 were synthesized by these procedures and characterized for the first time. These new isomers were assayed as inhibitors of glycosidases. As a result, ( )-hyacinthacine A2 demonstrated to be a good inhibitor of a-d-glucosidase from rice whereas the natural enantiomer, hyacinthacine A2, was not. Moreover, a new family of inhibitors of a-l-rhamnosidase was uncovered.Financial support from the Spanish MEC (CTQ2005–25182- E, CTQ2006–01080 and CTQ2006–01345/BQU), La Marat5 de TV3 foundation (Ref: 050931) and Generalitat de Catalunya DURSI 2005-SGR-00698 is acknowledged. J. Calveras acknowledges the CSIC UAs pre-doctoral scholarship programs.Peer reviewe

Influence of N-amino protecting group on aldolase-catalyzed aldol additions of dihydroxyacetone phosphate to amino aldehydes

9 pages, 3 scheme, 1 figure, 1 table.-- Printed version published Mar 13, 2006.This work examines the influence of N-protecting groups on the conversion and stereoselectivity of dihydroxyacetone phosphate (DHAP) dependent aldolase-catalyzed aldol additions of DHAP to N-protected-3-aminopropanal. Phenylacetyl-(PhAc-), tert-butyloxycarbonyl- (tBoc-) and fluoren-9-ylmethoxycarbonyl- (Fmoc-)-3-aminopropanal were evaluated as substrates for d-fructose 1,6-bisphosphate aldolase from rabbit muscle (RAMA), and l-rhamnulose-1-phosphate aldolase (RhuA) and l-fuculose-1-phosphate aldolase (FucA), both from Escherichia coli. Using PhAc and tBoc ca. 70% conversions to the aldol adduct were achieved, whereas Fmoc gave maximum conversions of ca. 25%. The stereoselectivity of the DHAP-aldolases did not depend on the N-protected-3-aminopropanal derivative. Moreover, inversion of FucA stereoselectivity relative to that obtained with the natural l-lactaldehyde was observed. Both N-PhAc and tBoc adduct product derivatives were successfully deprotected by penicillin G acylase (PGA)-catalyzed hydrolysis at pH 7 and by treatment with aqueous TFA (6% v/v), respectively. However, the corresponding cyclic imine sugars could not be isolated, presumable due to the presence of a highly reactive primary amine and a keto group in the molecule, which lead to a number of unexpected reactions.Financial support from the Spanish C.I.C.Y.T. (PPQ2002-04625-CO2-01 and BQU2003-01677) is acknowledged. J.C. acknowledges the CSIC for the I3P predoctoral scholarship. We are thankful to Prof. Josep López-Santín and the team of the Chemical Engineering Department at the Universitat Autònoma de Barcelona for the supply of FucA aldolase.Peer reviewe

Introducció a les Xarxes Telemàtiques: problemes amb solució

Col·lecció de problemes resolts d'IXT (Introducció a les Xarxes Telemàtiques) amb solució, 1er curs dels graus de l'ETSETB.2022/20232n quadrimestr

Introducció a les Xarxes Telemàtiques: problemes amb solució

Col·lecció de problemes resolts d'IXT (Introducció a les Xarxes Telemàtiques) amb solució, 1er curs dels graus de l'ETSETB.2022/20232n quadrimestr