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Combination of system and molecular biology approaches for discovery and characterization of plant enzymes for industrial processes



Funding: European Regional Development Fund (ERDF) “On Implementation of Activity “Post-doctoral Research Aid” of the Specific Aid Objective 1.1.1 “To increase the research and innovative capacity of scientific institutions of Latvia and the ability to attract external financing, investing in human resources and infrastructure” of the Operational Programme “Growth and Employment”

Project Title:Combination of system and molecular biology approaches for discovery and characterization of plant enzymes for industrial processes”

 Project Nr.

Period: 36 month (1st January 2019 – 31 st December 2021)

Project costs: 133 806,00 EUR

Project implementer: Dr. PhD. Paulius Lukas Tamošiūnas


The color was recognized to be the third most important aspect of food purchase choice, and there is a substantial lack of natural blue color pigments in the market. Several artificial colorants, including Blue No. 1 and Blue No. 2, were suggested to be causing hyperactivity in children and allergic reactions (Kobylewski, S., & Jacobson, M. F. (2010)). In recent years, the market of the food colors industry has rapidly increased and it is expected to continue growing 10% to 15% annually (Institute of Food Technologists. 2016) Therefore there is a considerable commercial interest in achieving strong blue food grade preparations and to improve the stability of anthocyanins such that they can be used as industrially-reliable, stable natural colorants. Anthocyanins have been approved for use in foods, in Europe with the label E163 (Cortez, R., Luna-Vital, D. A., Margulis, D., & Gonzalez de Mejia, E. (2017)).

Of particular importance for the use of anthocyanins as colorants in food, additives are the nature of the side-chain modification. Since glycosylation of C3 occurs on all naturally-accumulating anthocyanins and increases the stability of the pigment. Additionally, the sugar residues of anthocyanins are often acylated with aromatic acids (p-coumaric, caffeic, ferulic, sinapic, gallic or p-hydroxybenzoic acids) or aliphatic acids (malonic, acetic, malic, succinic, tartaric and oxalic acids). Aromatic acylation of anthocyanins increases their stability in solution by promoting self-association and/or the intramolecular stacking of the acyl groups onto the chromophore (intramolecular copigmentation). In some plants, the acyl groups are themselves glycosylated. Some complex anthocyanins have alternating glycosyl and\ acyl groups, which lead to further increased color stability in solution. Aromatic acylation additionally shifts the color of the pigments towards the blue (bathochromic shift in λmax), and some of the most intense blue colors of flowers, such as those found in morning glory and lobelia, are conferred by polyacylated (aromatic) anthocyanins. Anthocyanins having a dihydroxy substitution on their B-ring (cyanidin, delphinidin and petunidin derivatives) bind metal ions such as Fe3+, Mg2+, Ca2+, and Al3+. Finally, co-pigmentation and metal binding can combine to produce stable blue color. In the so-called metalloanthocyanins, three anthocyanin chromophores are bound to the metal ion and three flavone or flavonol molecules are inserted between them by p-stacking interactions.

Information published 02.01.2019.


Progress of the project:

1 January 2019 – 31 March 2019

During the initiation phase of the project current state of the biotechnological production of the anthocyanine and other blue-colored natural colorants was reviewed. A putative metabolic pathway to establish an efficient production of core anthocyanine species in yeast S. cerevisiae was designed with four phenolic acids as possible precursors. The stability, inhibition on the growth of various yeast strains, as well as the uptake to the cell of these four precursors were evaluated in MS, spectroscopical and growth assays. The vectors for recombinant production of plantal origin 4-coumaric acid-CoA ligase, chalcone synthase 1, chalcone isomerase, and dihydroflavonol 4-reductase in yeast were prepared. 

Information published 29.03.2019.


Progress of the project: 

1 April 2019 – 30 June 2019

The expression of recombinant 4-coumaric acid-CoA ligase, chalcone synthase 1, chalcone isomerase, and dihydroflavonol 4-reductase genes in S. cerevisiae was evaluated and the stability of the proteins was confirmed. The vectors for recombinant production of plantal origin dihydroflavonol 4-reductase and UDP-glucose flavonoid 3-O-glucosyltransferase in yeast were prepared. The platform for multiple gene expression in yeast was designed for metabolic pathway construction in yeast S. cerevisiae.

Information published 28.06.2019.

Mājas lapas izstrādi finansēja ERAF aktivitātes projekts Nr. 2010/0196/2DP/ "Latvijas biomedicīnas pētījumu integrācija Eiropas zinātnes telpā".