Development of Supporting Software for Driving School

This thesis is focused on design and implementation web application it based on the requirements and analysis of the current situation using technologies PHP and MySQL. It is specification system for planning rides, final exams and related functions in driving school

MOESM1 of Image-based methods for phenotyping growth dynamics and fitness components in Arabidopsis thaliana

Additional file 1. ImageJ macro used to extract rosette shape descriptors from top-view tray or pot images

Application of an Acyl-CoA Ligase from <i>Streptomyces aizunensis</i> for Lactam Biosynthesis

ε-Caprolactam and δ-valerolactam are important commodity chemicals used in the manufacture of nylons, with millions of tons produced annually. Biological production of these highly valued chemicals has been limited due to a lack of enzymes that cyclize ω-amino fatty acid precursors to corresponding lactams under ambient conditions. In this study, we demonstrated production of these chemicals using ORF26, an acyl-CoA ligase involved in the biosynthesis of ECO-02301 in <i>Streptomyces aizunensis</i>. This enzyme has a broad substrate spectrum and can cyclize 4-aminobutyric acid into γ-butyrolactam, 5-aminovaleric acid into δ-valerolactam and 6-aminocaproic acid into ε-caprolactam. Recombinant <i>E. coli</i> expressing ORF26 produced valerolactam and caprolactam when 5-aminovaleric acid and 6-aminocaproic acid were added to the culture medium. Upon coexpressing ORF26 with a metabolic pathway that produced 5-aminovaleric acid from lysine, we were able to demonstrate production of δ-valerolactam from lysine

Alteration of Polyketide Stereochemistry from <i>anti</i> to <i>syn</i> by a Ketoreductase Domain Exchange in a Type I Modular Polyketide Synthase Subunit

Polyketide natural products have broad applications in medicine. Exploiting the modular nature of polyketide synthases to alter stereospecificity is an attractive strategy for obtaining natural product analogues with altered pharmaceutical properties. We demonstrate that by retaining a dimerization element present in LipPks1+TE, we are able to use a ketoreductase domain exchange to alter α-methyl group stereochemistry with unprecedented retention of activity and simultaneously achieve a novel alteration of polyketide product stereochemistry from <i>anti</i> to <i>syn</i>. The substrate promiscuity of LipPks1+TE further provided a unique opportunity to investigate the substrate dependence of ketoreductase activity in a polyketide synthase module context

Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in <i>Saccharomyces cerevisiae</i>

<div><p>Biological synthesis of pharmaceuticals and biochemicals offers an environmentally friendly alternative to conventional chemical synthesis. These alternative methods require the design of metabolic pathways and the identification of enzymes exhibiting adequate activities. Cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates are natural metabolites which possess beneficial activities for human health, and the search is expanding for novel derivatives that might have enhanced biological activity. For example, biosynthesis in <i>Dianthus caryophyllus</i> is catalyzed by hydroxycinnamoyl/benzoyl-CoA:anthranilate <i>N</i>-hydroxycinnamoyl/ benzoyltransferase (HCBT), which couples hydroxycinnamoyl-CoAs and benzoyl-CoAs to anthranilate. We recently demonstrated the potential of using yeast (<i>Saccharomyces cerevisiae</i>) for the biological production of a few cinnamoyl anthranilates by heterologous co-expression of 4-coumaroyl:CoA ligase from <i>Arabidopsis thaliana</i> (4CL5) and HCBT. Here we report that, by exploiting the substrate flexibility of both 4CL5 and HCBT, we achieved rapid biosynthesis of more than 160 cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates in yeast upon feeding with both natural and non-natural cinnamates, dihydrocinnamates, benzoates, and anthranilates. Our results demonstrate the use of enzyme promiscuity in biological synthesis to achieve high chemical diversity within a defined class of molecules. This work also points to the potential for the combinatorial biosynthesis of diverse and valuable cinnamoylated, dihydrocinnamoylated, and benzoylated products by using the versatile biological enzyme 4CL5 along with characterized cinnamoyl-CoA- and benzoyl-CoA-utilizing transferases.</p></div

Siliques_aut

This file contains the skeleton parameters extracted from image analysis of each plant inflorescence (see related papers). This file is used to automatically estimated fruit number from pictures of inflorescences

expanding_rosette_area

This file contains rosette area (cm2) and shape descriptors for all plants (451 accessions with 2 replicates) grown until the end of the life cycle. Data were extracted from image analysis (see related papers) during the 25 first days after vernalization to estimate rosette dry mass and growth dynamics with a modelling approach

Detection of <i>N</i>-(4’-hydroxydihydrocinnamoyl)-anthranilate (DHavnD) from the recombinant yeast culture medium.

<p>Representative ESI-MS spectra were obtained after LC-TOF MS analysis of <b>(A)</b> the culture medium of recombinant yeast incubated with anthranilate and 4-hydroxydihydrocinnamate, and <b>(B)</b> a DHavnD standard solution.</p

harvest_16d

This file file contains rosette area (cm2) and shape descriptors, as well as rosette dry mass (mg) and leaf dry mass per area (LMA, g m-2) measured on 817 individuals (n = 2 per accession) by image analysis, plant dissection and weighing. Plants have been harvested 16 days after the end of vernalization (see related papers for details)

harvest_16d

This file file contains rosette area (cm2) and shape descriptors, as well as rosette dry mass (mg) and leaf dry mass per area (LMA, g m-2) measured on 817 individuals (n = 2 per accession) by image analysis, plant dissection and weighing. Plants have been harvested 16 days after the end of vernalization (see related papers for details)