Bimodal substrate binding in the active site of the glycosidase BcX.

Bacillus circulans xylanase (BcX) from the glycoside hydrolase family 11 degrades xylan through a retaining, double-displacement mechanism. The enzyme is thought to hydrolyze glycosidic bonds in a processive manner and has a large, active site cleft, with six subsites allowing the binding of six xylose units. Such an active site architecture suggests that oligomeric xylose substrates can bind in multiple ways. In the crystal structure of the catalytically inactive variant BcX E78Q, the substrate xylotriose is observed in the active site, as well as bound to the known secondary binding site and a third site on the protein surface. Nuclear magnetic resonance (NMR) titrations with xylose oligomers of different lengths yield nonlinear chemical shift trajectories for active site nuclei resonances, indicative of multiple binding orientations for these substrates for which binding and dissociation are in fast exchange on the NMR timescale, exchanging on the micro- to millisecond timescale. Active site binding can be modeled with a 2 : 1 model with dissociation constants in the low and high millimolar range. Extensive mutagenesis of active site residues indicates that tight binding occurs in the glycon binding site and is stabilized by Trp9 and the thumb region. Mutations F125A and W71A lead to large structural rearrangements. Binding at the glycon site is sensed throughout the active site, whereas the weak binding mostly affects the aglycon site. The interactions with the two active site locations are largely independent of each other and of binding at the secondary binding site.

Eco-friendly approach to improve traits of winter wheat by combining cold plasma treatments and carbonization of subtropical biomass waste.

This study aims to improve the quality and quantity of winter wheat by using the potential of combining the use of cold plasma and waste biorefinery products for improving wheat yield. Plasma was applied by a radio frequency (RF) plasma reactor operated with air for 180 s and 50 W. The waste biorefinery products, including pyroligneous acid, biochar, and azolla compost, were used as plant nutrition. The effects of cold plasma treatment and waste biorefinery products were determined by measuring plant photosynthesis, grain yield, and content of chlorophyll, carotenoids, anthocyanin, protein, and starch. The experiment was conducted during the cropping seasons 2016-18 in a randomized complete block design with four replications. The combination of cold plasma and pyroligneous acid increased the grain yield up to 40.0%. The photosynthesis rate was improved up to 39.3%, and total chlorophyll content up to 48.3% in both years. Seed plasma treatment combined with biochar application increased the starch content by 36.8%. Adding azolla compost increased the protein content by 35.4%. Using seed plasma treatment with biochar increased the microbial biomass carbon by 16.0%. The application of plasma and azolla compost increased the microbial biomass nitrogen by 29.0%.

Amelioration of Photosynthesis and Quality of Wheat under Non-thermal Radio Frequency Plasma Treatment.

Plasma treatment is recognized as a suitable technology to improve germination efficiency of numerous seeds. The objective of this paper is to demonstrate whether cold air plasma can change the quality and quantity of wheat yield. Effects of cold plasma treatment on wheat (Pishgam variety) yield were studied by a randomized complete block design experiment at the Faculty of Agriculture of Tarbiat Modarres University, Iran, during 2015-17. Seeds were pre-treated with 80 W of cold plasma at four levels of time, 60, 120, 180 and 240 seconds. Plasma effects on yield and quality of wheat were determined by measuring plant photosynthesis, grain yield, biological yield, 1000-grain weight, total chlorophyll, carotenoid, anthocyanin, protein and starch content. Results showed that plasma treatments had positive effects on wheat characteristics, and treatment of 180 s had the highest stimulatory effect. In both years, cold plasma increased grain yield at 180 s, but decreased it at 240 s compared with control. The rate of plant photosynthesis, grain yield, 1000-grain weight, carotenoid and anthocyanin were enhanced at 180 s. The starch content and grain protein were enhanced at 120 s cold plasma application compared with control.