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The article explains the preparation of photocatalytic layers based on geopolymer. It notes that metakaolinite-based geopolymer composites with 10 wt% of TiO2 powders are formed when metakaolinite and sodium silicate solution is mixed with TiO2 powders. Using selected methods such as X-ray diffraction and Barrett-Joiner-Halenda (BJH) method, prepared materials in form of layers on steel plates are investigated.

Popularity of hyaluronan (HA) in the cosmetics and pharmaceutical industries, led to the investigation and development of new HA-based materials, with enzymes playing a key role. Beta-D-glucuronidases catalyze the hydrolysis of a beta-D-glucuronic acid residue from the non-reducing end of various substrates. However, lack of specificity towards HA for most beta-D-glucuronidases, in addition to the high cost and low purity of those active on HA, have prevented their widespread application. In this study, we investigated a recombinant beta-glucuronidase from Bacteroides fragilis (rBfGUS). We demonstrated the rBfGUS's activity on native, modified, and derivatized HA oligosaccharides (oHAs). Using chromogenic beta-glucuronidase substrate and oHAs, we characterized the enzyme's optimal conditions and kinetic parameters. Additionally, we evaluated rBfGUS's activity towards oHAs of various sizes and types. To increase reusability and ensure the preparation of enzyme-free oHA products, rBfGUS was immobilized on two types of magnetic macroporous bead cellulose particles. Both immobilized forms of rBfGUS demonstrated suitable operational and storage stabilities, and their activity parameters were comparable to the free form. Our findings suggest that native and derivatized oHAs can be prepared using this bacterial beta-glucuronidase, and a novel biocatalyst with enhanced operational parameters has been developed with a potential for industrial use., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Acknowledgements The research was supported by projects of Charles University (SVV 260636 and Cooperatio 207032-3 Immunity and Infection), (Copyright © 2023 Elsevier Ltd. All rights reserved.)

This paper describes the development of a straightforward method for site-directed gene mutagenesis in Streptococcus zooepidemicus, inspired by the mechanism of natural competence regulated by ComX in other streptococci. An alternative sigma factor comX gene was overexpressed from a plasmid in S. zooepidemicus and electrocompetent cells were prepared. As proof of concept, a DNA cassette with two targeting regions flanking a kanamycin resistance gene was spliced in an overlap extension PCR and electroporated. The cassette was then integrated in the genomic DNA by homologous recombination. Next, the gene SeseC_00180 (fibrinogen- and Ig-binding protein precursor) was selected as target for markerless gene deletion and the impact of its loss on the resulting hyaluronan production was determined. The new method of site-directed mutagenesis is significant because it is not necessary to clone the DNA cassette in an auxiliary vector, electroporating it in S. zooepidemicus cells is enough, which allows to bypass the problems with hard to clone DNA sequences and speeds up the whole process of mutation generation in S. zooepidemicus., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Human enterokinase (enteropeptidase, rhEP), a serine protease expressed in the proximal part of the small intestine, converts the inactive form of trypsinogen to active trypsin by endoproteolytic cleavage. The high specificity of the target site makes enterokinase an ideal tool for cleaving fusion proteins at defined cleavage sites. The mature active enzyme is comprised of two disulfide-linked polypeptide chains. The heavy chain anchors the enzyme in the intestinal brush border membrane, whereas the light chain represents the catalytic enzyme subunit. The synthetic gene encoding human enteropeptidase light chain with His-tag added at the C-terminus to facilitate protein purification was cloned into Pichia pastoris expression plasmids under the control of an inducible AOX1 or constitutive promoters GAP and AAC. Cultivation media and conditions were optimized as well as isolation and purification of the target protein. Up to 4 mg/L of rhEP was obtained in shake-flask experiments and the expression level of about 60-70 mg/L was achieved when cultivating in lab-scale fermentors. The constitutively expressing strains proved more efficient and less labor-demanding than the inducible ones. The rhEP was immobilized on AV 100 sorbent (Iontosorb) to allow repeated use of enterokinase, showing specific activity of 4U/mL of wet matrix., (Copyright © 2011 Elsevier B.V. All rights reserved.)