An Arabidopsis thaliana gene encoding a homologue of the potato alpha-glucan, water dikinase GWD, previously known as R1, was identified by screening genome and named AtGWD3. The AtGWD3 cDNA isolated, heterologously expressed protein purified to apparent homogeneity determine enzymatic function. In contrast GWD protein, primarily catalysed phosphorylation at C-3 position glucose unit preferably pre-phosphorylated amylopectin substrate with long side chains. mutant, termed Atgwd3,...

Starch granule types from a variety of botanical sources were selected to represent differences in crystalline polymorph, amylose and phosphate content, amylopectin chain length distribution. Equimolar labeling starch molecules with the fluorophore 8-amino-1,3,6-pyrenetrisulfonic acid (APTS) was used construct detailed map distribution within by confocal laser scanning microscopy (CLSM) analysis. Medium- high-resolution electron (SEM) provide images surface structures. By using combined...

The use of cold-active enzymes has many advantages, including reduced energy consumption and easy inactivation. ikaite columns SW Greenland are permanently cold (4-6°C) alkaline (above pH 10), the microorganisms living there their adapted to these conditions. Since only a small fraction total microbial diversity can be cultured in laboratory, combined approach involving functional screening strain collection metagenomic library was undertaken for discovery novel from columns. A with 322...

ABSTRACT Potatoes are cultivated in southwest Greenland without the use of pesticides and with limited crop rotation. Despite fact that plant-pathogenic fungi present, no severe-disease outbreaks have yet been observed. In this report, we document a potato soil at Inneruulalik southern is suppressive against Rhizoctonia solani Ag3 uncover antifungal mechanism highly potent biocontrol bacterium, Pseudomonas fluorescens In5, isolated from soil. A combination molecular genetics, genomics,...

Marine microbes are a rich source of enzymes for the degradation diverse polysaccharides. Paraglaciecola hydrolytica S66T is marine bacterium capable hydrolyzing polysaccharides found in cell wall red macroalgae. In this study, we applied an approach combining genomic mining with functional analysis to uncover potential produce hydrolysis complex A special feature P. presence large region harboring array carbohydrate-active (CAZymes) notably agarases and carrageenases. Based on first...

Starch is the major carbohydrate reserve in plants, and degraded for growth at night. breakdown requires reversible glucan phosphorylation granule surface by novel dikinases phosphatases. The dual-specificity phosphatase starch excess 4 (SEX4) required desphosphorylation; however, regulation of enzymatic activity SEX4 not well understood. We show that switches between reduced (active) oxidized (inactive) states, suggesting redox-regulated. Although only partial reactivation was achieved...

Starch biosynthesis in Arabidopsis thaliana is strictly regulated. In leaf extracts, starch synthase 1 (AtSS1) responds to the redox potential within a physiologically relevant range. This study presents data testing two main hypotheses: 1) that specific thiol-disulfide exchange AtSS1 influences its catalytic function 2) each conserved Cys residue has an impact on catalysis. Recombinant versions carrying combinations of cysteine-to-serine substitutions were generated and characterized vitro....

The submarine ikaite columns located in the Ikka Fjord Southern Greenland represent a unique, permanently cold (less than 6°C) and alkaline (above pH 10) environment are home to microbial community adapted these extreme conditions. bacterial archaeal inhabiting surrounding fjord was characterised by high-throughput pyrosequencing of 16S rRNA genes. Analysis structure revealed presence diverse community, both column interior at surface, very few archaea. A clear difference overall taxonomic...

AbstractA field survey of local traditional food habits in northern Italy revealed that children Carnia have traditionally eaten sweet ingluvies (the crop) from day flying moths the genus Zygaena and its mimic, Syntomis. These are brightly colored, all species contain cyanogenic glucosides, which release toxic hydrogen cyanide upon degradation. The presence glucosides larvae imagos (adults) as well Syntomis was investigated using liquid chromatography-mass spectrometry. sugar content...

The family 20 carbohydrate‐binding module (CBM20) of the Arabidopsis starch phosphorylator glucan, water dikinase 3 (GWD3) was heterologously produced and its properties were compared to CBM20 from a fungal glucoamylase (GA). GWD3 has 50‐fold lower affinity for cyclodextrins than that GA. Homology modelling identified possible structural elements responsible this weak binding intracellular CBM20. Differential fluorescein‐labelled GA modules granules in vitro demonstrated by confocal laser...

Starch-binding domains are noncatalytic carbohydrate-binding modules that mediate binding to granular starch. The starch-binding from the module family 45 (CBM45, http://www.cazy.org) found as N-terminal tandem repeats in a small number of enzymes, primarily photosynthesizing organisms. Isolated representatives each two classes enzyme carrying CBM45-type domains, Solanum tuberosumα-glucan, water dikinase and Arabidopsis thaliana plastidial α-amylase 3, were expressed recombinant proteins...

Sulfated fucans, often denoted as fucoidans, are highly variable cell wall polysaccharides of brown algae, which possess a wide range bioactive properties with potential pharmaceutical applications. Due to their complex architecture, the structures algal fucans have until now only been partly determined. Enzymes capable hydrolyzing sulfated may allow specific release defined oligosaccharides and serve tool for structural elucidation walls. Currently, such enzymes include few hydrolases...

The fungal pathogen Blumeria graminis f. sp. hordei develops on the barley leaf via distinct, morphologically well-defined stages. After landing a host plant, conidia rapidly germinate to form primary germ tube. Subsequently, an appressorial tube emerges from conidium and differentiates appressorium which penetration of cell wall is attempted. We have used serial analysis gene expression provide measurement messenger RNA contents in ungerminated conidia, during conidial germination,...

Few studies to date report the transcriptional response of biocontrol bacteria toward phytopathogens. In order gain insights into potential mechanism underlying antagonism antimicrobial producing strain P. fluorescens In5 against phytopathogens Rhizoctonia solani and Pythium aphanidermatum, global RNA sequencing was performed.Differential gene expression profiling in either R. or aphanidermatum investigated using transcriptome (RNA-seq). Total isolated from single bacterial cultures...

The Ikka Fjord (SW Greenland) harbors a unique microbial habitat in the form of several hundred submarine tufa columns composed ikaite, special hexahydrate calcium carbonate that precipitates when alkaline phosphate- and carbonate-enriched spring water seeping out sea floor meets cold seawater. While heterotrophic microbes have been isolated from columns, activity boundary conditions for growth ikaite remained unexplored. We present first detailed situ characterization physico-chemical...

Starch phosphorylation catalysed by the α-glucan, water dikinases (GWD) has profound effects on starch degradation in plants. The Arabidopsis thaliana genome encodes three isoforms of GWD, two which are localized chloroplast and involved transient starch. third isoform, termed AtGWD2 (At4g24450), was heterologously expressed purified shown to have a substrate preference similar potato GWD. Analyses null mutants did not reveal any differences growth or sugar levels, when compared wild type....