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We improved an already existing cytochrome c expression system to a reliable, tightly controllable one to achieve a higher expression yield for single cysteine mutants of horse cytochrome c. The protein is heterologously overexpressed in E. coli together with the maturation coordinating enzyme heme lyase from yeast. Various plasmid constructs and host strains were tested for protein expression yield and routinely around 35 mg/L yield was achieved, which is a good result for a post-translationally modified enzyme. The purpose of producing cysteine mutants is to position accessible cysteine residues on the surface of cytochrome c which can be labeled with a photoactive redox dye, 8-thiouredopyrene-1,3,6-trisulfonate, TUPS. TUPS labeled proteins have been used for intramolecular and intermolecular electron transfer measurements. Here, we initiate the photoreduction of cytochrome c oxidase, the natural electron acceptor partner of cytochrome c by an appropriate cytochrome c mutant labeled with TUPS. The electron transfer from cytochrome c to the first cytochrome oxidase redox cofactor, copper A, is shown to be very fast.

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Acta Biologica Hungarica
Authors: Mónika Magony, Ildikó Kákonyi, Anna Gara, P. Rapali, Katalin Perei, K. Kovács, and G. Rákhely

A bacterium capable to grow on sulfanilic acid as sole carbon, nitrogen and sulfur source has been isolated. A unique feature of this strain is that it contains the full set of enzymes necessary for the biodegradation of sulfanilic acid. Taxonomical analysis identified our isolate as Sphingomonas subarctica SA1 sp. The biodegradation pathway of sulfanilic acid was investigated at the molecular level. Screening the substrate specificity of the strain disclosed its capacity to degrade six analogous aromatic compounds including p -aminobenzoic acid. Moreover, the strain was successfully used for removal of oil contaminations. S. subarctica SA1 seemed to use distinct enzyme cascades for decomposition of these molecules, since alternative enzymes were induced in cells grown on various substrates. However, the protein patterns appearing upon induction by sulfanilic acid and sulfocatechol were very similar to each other indicating common pathways for the degradation of these substrates. Cells grown on sulfanilic acid could convert p -aminobenzoic acid to some extent and vice versa. Two types of ring cleaving dioxygenases were detected in the cells grown on various substrates: one preferred protocatechol, while the other had higher activity with sulfocatechol. This latter enzyme, named as sulfocatechol dioxygenase was partially purified and characterized.

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Acta Alimentaria
Authors: Sz. Gy. Szerencsés, S. Beszédes, Zs. László, G. Veréb, D. Szalay, Zs. Hovorkáné Horváth, C. Hodúr, G. Rákhely, and Sz. Kertész

Abstract

In this study, statistical analysis was performed to investigate the influence of operational parameters based on experimental results. The research aimed to know whether a long-term discontinuous module vibration operation is possible without adversely affecting filtration efficiency. Polymer membranes were compared by surface characteristics with contact angle measurements and selected for further membrane filtration experiments for dairy wastewater treatment. The effect of the main operational parameters, membrane module vibration amplitude (A vibr.) and transmembrane pressure (TMP), was investigated using a vibratory shear enhanced processing (VSEP) module with ultrafiltration (UF) and nanofiltration (NF) membranes. Components of the permeates, including chemical oxygen demand (COD), were measured, and membrane rejections were calculated to compare with threshold limit values. The results suggest that proper combination of A vibr. and TMP could mitigate membrane fouling. However, discontinuous module vibration resulted in more clogged membrane with lower fluxes, but slightly higher rejections. We conclude that investigating the significance of operational parameters is necessary for a wider, more energy and environment-friendly application in wastewater treatment.

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