Studies of the impact of terahertz radiation on living objects present a significant interest since its use for security systems is currently considered promising. We studied the non-thermal impact of terahertz radiation on E. coli/pKatG-gfp biosensor cells. The Novosibirsk free electron laser (NovoFEL), which currently has the world's highest average and peak power, was used as the source of terahertz radiation. We demonstrated that exposure to terahertz radiation at the wavelengths of 130, 150, and 200 µm and a power of 1.4 W/cm(2) induces changes in green fluorescent protein (GFP) fluorescence values and thus induces the expression of GFP in E. coli/pKatG-gfp biosensor cells. Possible mechanisms of the E. coli response to non-thermal exposure to terahertz radiation are discussed.
The effective utilisation of available genetic resources of related species is essential for successful crops breeding and maintaining genetic variability within crops. Bread wheat, the basic cultivated wheat species, is an amphiploid (2n = 6x = 42) and, therefore, the production of new synthetic amphiploids using genomes of related species should reduce the difficulties caused by direct crossings, for example, between hexaploid wheat and diploid relatives. Hence, exploiting synthetic amphiploids is an effective and rapid way of introgressing desirable traits from related species into cultivated wheats. Some of the artificial amphiploids that already exist were produced 80 years ago. Yet little work has been done to highlight potential contamination and/or genetic changes during their conservation by genebanks. Thus, we utilised the electrophoresis of wheat endosperm storage proteins (gliadins) to check such amphiploid authenticity, and also where differences had been previously observed between synthetic wheat amphiploids.In addition, we checked putative amphiploid accessions where Triticum timopheevii (GGA t A t ) was recorded as one of the parents. A synthetic species, T. timococcum produced by Kostov, together with a natural T. zhukovskyi found in Georgia (the former Soviet Union) were revealed to be identical according to our assays. The existence of several T. kiharae accessions independently produced by different authors was confirmed, and they exhibited polymorphism for a number of traits, including spike characters (awning, hairy glumes) and growth habit (spring vs. winter). The effective conservation of artificial amphiploids in genebanks is discussed.
Xylanases (EC 3.2.1.8) hydrolyze the hemicellulose of plant cell walls. Xylanases are used in the food and paper industries and for bioconversion of lignocellulose to biofuel. In this work, the producer-strain with four copies of the xAor xylanase gene was organized in two tandem copies for optimal expression in Komagataella phaffii T07 yeast. The secreted 35 kDa xylanase was purified from culture medium by gel filtration on Sephadex G-25 and anion exchange chromatography on DEAE-Sepharose 6HF. Tryptic peptides of the recombinant enzyme were analyzed by liquid chromatography-tandem mass spectrometry where the amino acid sequence corresponded to Protein Accession # O94163 for Endo-1,4-beta-xylanase from Aspergillus oryzae RIB40. The recombinant xylanase was produced in a bioreactor where the secreted enzyme hydrolyzed oat xylane with an activity of 258240 IU/mL. High activity in the culture medium suggested xylanase could be used for industrial applications without being purified or concentrated. The pH optimum for xylanase xAor was 7.5, though the enzyme was active from pH 2.5 to pH 10. Xylanase was active at temperatures from 35 °C to 85 °C with a maximum at 60 °C. In conclusion, this protocol yields soluble, secreted xylanase suitable for industrial scale production.
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