About
13
Publications
766
Reads
How we measure 'reads'
A 'read' is counted each time someone views a publication summary (such as the title, abstract, and list of authors), clicks on a figure, or views or downloads the full-text. Learn more
72
Citations
Introduction
Current institution
Publications
Publications (13)
The 3'X tail is a functionally essential 98-nt sequence located at the 3'-end of the hepatitis C virus (HCV) RNA genome. The domain contains two absolutely conserved dimer linkage sequence (DLS) and k nucleotide segments involved in viral RNA dimerization and in a distal base-pairing interaction with stem-loop 5BSL3.2, respectively. We have previou...
The 3'X domain is a 98-nucleotide region located at the 3' end of hepatitis C virus genomic RNA that plays essential functions in the viral life cycle. It contains an absolutely conserved, 16-base palindromic sequence that promotes viral RNA dimerization, overlapped with a 7-nucleotide tract implicated in a distal contact with a nearby functional s...
The 3'X domain of hepatitis C virus is a strongly conserved structure located at the 3' terminus of the viral genomic RNA. This domain modulates the replication and translation processes of the virus in conjunction with an upstream 5BSL3.2 stem-loop, and contains a palindromic sequence that facilitates RNA dimerization. Based on nuclear magnetic re...
A homologue of the Escherichia coli penicillin acylase is encoded in the genomes of several thermophiles, including in different Thermus thermophilus strains. Although the natural substrate of this enzyme is not known, this acylase shows a marked preference for penicillin
K over penicillin G. Three-dimensional models were created in which the catal...
Penicillin acylases (PACs) are enzymes of industrial relevance in the manufacture of β-lactam antibiotics. Development of a PAC with a longer half-life under the reaction conditions used is essential for the improvement of the operational stability of the process. A gene encoding a homologue to Escherichia coli PAC was found in the genome of the th...
Optimum pH of HIS6::TthPAC. The TthPAC enzymatic activity was assayed at 65°C in the presence of 2.5 mM PenK and in 20 mM Britton-Robinson buffer at pH 3, 4, 5, 6, 7, 8 or 9.
Comparison of the kinetic data of TthPAC with other penicillin acylases. Data were measured at 40°C unless otherwise stated.
Chaperone elimination after co-expression with HIS6:: Tth PAC. Tth PAC was co-expressed with GroEL/ES in E. coli BL21 cells. Total soluble protein fraction was incubated in the absence (lanes 1–4) or in the presence of 5 mM ATP/10 mM Cl2Mg (lanes 5–8) [41] for 2 h at 4°C. In order to separate the Tth PAC from the detached GroEL/ES a 0-, 30-, 45- or...
Heterologous expression of the chimeric protein SpEco-PACTthin E. coli cells.TthPAC α- and β-subunit were immunodetected in the cytoplasmic fraction (upper and middle panels) and in the periplasmic fraction (lower pannel) of E. coli cells from BL21 strain(1, A and G); Rosetta-gami2 strain (2, B and H); BL21 strain co-expressing GroEL/ES and trigger...
Sequence alignment of PKAs and characterized PGAs. TthPKA, Thermus thermophilus HB27 PKA [TTC1972]; AutaPKA, Actinoplanes utahensis PKA [P29958]; SlavPKA, Streptomyces lavendulae PKA [AY611030]; EcoPGA, Escherichia coli PGA [P06875]; AfaePGA, Alcaligenes faecalis [ADD11517]; PretPGA, Providencia rettgeri PGA [AAP86197]; KcitPGA, Kluyvera citrophila...
Thermal inactivation course of TthPAC. Solutions containing 0.08 mg/ml of purified TthPAC were incubated in 50 mM phosphate buffer pH 7.5 at 65 and 75°C. The residual hydrolytic activity was determined using 5 mM penicillin K as substrate.
Table S2. Primers used in this work.
