engleski

CONSTRUCTION OF Saccharomyces cerevisiae STRAINS FOR MYCOSPORINE-LIKE AMINO ACIDS PRODUCTION

Mycosporine-like amino acids are compounds known for their UV-absorbing properties and are found in numerous marine microorganisms which produce these secondary metabolites as a way of dealing with UV-induced stress. Their absorption maxima typically range from 310 to 360 nm. The aim of this thesis was to construct Saccharomyces cerevisiae strains with heterologous biosynthetic mycosporine-like amino acids pathway from cyanobacterium Anabaena variabilis stably integrated into chromosomes 10 and 11. Three plasmids containing the pathway from A. variabilis with corresponding constitutive promoters and terminators were constructed. The pathway was integrated into haploid and diploid S. cerevisiae strains and into their derivatives which express a heterologous pathway for xylose consumption so that the strains would be able to grow on medium supplemented with xylose, in addition to glucose. Plasmid constructs were verified by colony PCR and sequencing. Colony PCR was used to confirm vector integration into S. cerevisiae chromosomes and high performance liquid chromatography (HPLC) combined with mass spectrometry (LC-MS) was used to detect the produced compounds from the mycosporine-like amino acids pathway in constructed yeast strains.

mycosporine-like amino acids, xylose isomerase pathway, pentose phosphate pathway

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