The expression of heterologous proteins in Escherichia coli is strongly affected by codon bias. This phenomenon occurs when the usage mRNA coding for foreign protein differs from that bacterium. ribosome pauses upon encountering a rare and may detach mRNA, thereby yield reduced. Several bacterial strains have been engineered to overcome this effect. However, increased rate translation lead misfolding insolubilization. In order prove assumption, solubility several recombinant plants was...

EDITORIAL article Front. Microbiol., 08 July 2014Sec. Microbiotechnology Volume 5 - 2014 | https://doi.org/10.3389/fmicb.2014.00341

The zinc metalloenzyme β-lactamase II (βLII) from Bacillus cereus has been overexpressed in Escherichia coli as a fusion protein with glutathione-S-transferase, and the metal binding properties of recombinant βLII toward Zn(II) Co(II) have studied by fluorescence activity measurements. apoenzyme is able to bind two ion equivalents, which confer on its maximum enzymatic efficiency. enzyme partially active one equivalent. diCo(II) mixed Zn(II)Co(II) derivative were obtained probed electronic...

Plants constitute a source of novel phytotoxic compounds to be explored in searching for effective and environmentally safe herbicides. From previous screening plant extracts their phytotoxicity, dichloromethane extract Ammi visnaga (L.) Lam. was selected further study. Phytotoxicity-guided fractionation this yielded two furanochromones, khellin visnagin, which herbicidal activity had not been described before. Khellin visnagin were model species lettuce (Lactuca sativa) duckweed (Lemna...

Chloroplast ferredoxin-NADP⁺reductase has a 32,000-fold preference for NADPH over NADH, consistent with its main physiological role of NADP⁺ photoreduction <i>de novo</i> carbohydrate biosynthesis. Although it is distant from the 2′-phosphoryl group NADP⁺, replacement C-terminal tyrosine (Tyr³⁰⁸ in pea enzyme) by Trp, Phe, Gly, and Ser produced enzyme forms which NADH was decreased about 2-, 10-, 300-, 400-fold, respectively. Remarkably, case Y308S...

We have analyzed the interaction of DnaK and plant Hsp70 proteins with wild‐type ferredoxin‐NADP + reductase precursor (preFNR) mutants containing amino‐acid replacements in targeting sequence. Using an algorithm already developed [Rüdiger, S., Germeroth, L., Schneider‐Mergener, J. &amp; Bukau, B. (1997) EMBO 16 , 1501–1507] we observed that 75% 727 plastid contained at least one site high likelihood binding their transit peptides. Statistical analysis showed a decrease frequency within...

The ferredoxin-NADP+ oxidoreductase of spinach chloroplasts was purified from a Triton X-100 thylakoid extract closely associated with an intrinsic polypeptide 17.5 kDa. 17.5-kDa polypeptide-reductase complex differs soluble reductase in (a) its elution profile Affi-Gel blue column; (b) behavior isoelectric focusing electrophoresis; and (c) giving different immunoelectrophoretic arcs. diaphorase activity the showed same pH thylakoid-bound reductase. curve changed to form similar that after...

HSP100 proteins are molecular chaperones involved in protein quality control. They assist (un)folding, prevent aggregation, and thought to participate precursor translocation across membranes. Caseinolytic ClpC ClpD from plant chloroplasts belong the family. Their role has hitherto been investigated by means of physiological studies reverse genetics. In present work, we employed an vitro approach delve into structural functional characteristics ClpC2 Arabidopsis thaliana (AtClpC2 AtClpD)....

The catalytic mechanism proposed for ferredoxin-NADP+ reductase (FNR) is initiated by reduction of its flavin adenine dinucleotide (FAD) cofactor the obligatory one-electron carriers ferredoxin (Fd) or flavodoxin (Fld) in presence oxidized nicotinamide phosphate (NADP+). C-terminal tyrosine FNR, which stacks onto ring, modulates enzyme affinity NADP+/H, being removed from this stacking position during turnover to allow productive docking and hydride transfer. Due location at...

Endolysins are peptidoglycan hydrolases with promising use as environment-friendly antibacterials mainly when used topically. However, in general, endolysin expression is hampered by its low solubility. Thus, a critical point industrial production optimizing their expression, including improvement of solubility and recovery from cell extracts.

Ferredoxin-NADP(H) reductase (FNR) is a FAD-containing protein that catalyzes the reversible transfer of electrons between NADP(H) and ferredoxin or flavodoxin. This enzyme participates in redox-based metabolism plastids, mitochondria, bacteria. Plastidic plant-type FNRs are very efficient reductases supporting photosynthesis. They have strong preference for over NAD(H), consistent with main physiological role NADP+ photoreduction. In contrast, from organisms heterotrophic metabolisms...

Abstract Background Clp/Hsp100 chaperones are involved in protein quality control. They act as independent units or conjunction with a proteolytic core to degrade irreversibly damaged proteins. Clp from plant chloroplasts have been also implicated the process of precursor import, along Hsp70 chaperones. thought pull precursors transit peptides enter organelle. How identify their substrates and engage processing is not known. This information may lie position, sequence structure recognition...

Abstract An important characteristic of promoters used in recombinant protein production Escherichi coli is their inducibility a simple and cost‐effective manner. The IPTG inducible lac, tac , trc are powerful widely for basic research. However, the use large‐scale undesirable due to its high cost toxicity. mentioned above can also be induced by addition lactose, which has double role inducer carbon energy source. Nevertheless, this sugar industrial processes several drawbacks, result low...

The carboxyl-terminal region of plant ferredoxin-NADP+ reductases is formed by an invariant alpha-helix/loop/beta-strand, culminating in a conserved tyrosine that displays extensive interaction with the prosthetic group FAD. We have investigated potential role terminal reductase function, introducing mutations and deletions on pea overexpressed Escherichia coli. Replacement tryptophan, phenylalanine, serine, glycine resulted 2.2-, 2.0-, 22-, 302-fold reduction, respectively, kcat for...

Abstract The aziridinium of purified quinacrine mustard at 50 microM inactivates the bovine heart mitochondrial F1-ATPase with a pseudo-first order rate constant 0.07 min-1 pH 7.0 and 23 degrees C. An apparent Kd 27 for enzyme-reagent complex was estimated from dependence inactivation on concentration mustard. profile revealed that deprotonation group pKa about 6.7 is necessary inactivation. amount reagent incorporated into protein increased linearly extent Complete to occur when 3 mol were...