Аннотация и ключевые слова
Аннотация (русский):
The results of the analysis of DNA sequences encoding L-phenylalanine ammonia-lyase (PAL) synthesis, performed to obtain universal primers complementary to conserved regions of the pal gene, are presented in the article. The fragment of pal gene was amplified in organisms under study. Nucleotide sequence of the pal gene in microorganisms exhibiting L-phenylalanine ammonia-lyase activity was determined by DNA sequencing. The results of its comparison with the corresponding sequences of known species are presented. Phenotypic characteristics and biochemical properties of selected cultures were studied. An investigation aimed to choose a superproducer strain of L-phenylalanine ammonia-lyase was conducted. It was found that L-phenylalanine ammonia-lyase synthesis was the most active in the following strains: Aureobasidium pullulans Y863, Rhodosporidium infirmominiatum Y1569, Candida glabrata Y2813, Candida maltose Y242, Debaryomyces robertsiae Y3392, Rhodosporidium diobovatum Y1565, Rhodotorula lactose Y2770, Saccharomyces cerevisiae Y1127, Tilletiopsis washingtonensis Y1650, Torulopsis apicola Y566, Tremella foliacea Y1624, Rhodotorula rubra Y1193, and Debaryomyces castellii Y968. This allows recommending them for further research aimed to obtain the enzyme preparation of L-phenylalanine ammonia-lyase.

Ключевые слова:
L-phenylalanine ammonia-lyase, enzyme, pal gene, pigmental yeast, nucleotide sequence, amino acid sequence, phylogenetic tree


L-phenylalanine ammonia-lyase (PAL; EC catalyzes the reaction of reverse deamination of L-phenylalanine to trans-cinnamic acid and ammonia [1]. It is the key enzyme of phenylpropanoid metabo-lism in plant and fungi, where it is involved in biosynthesis of secondary metabolites (flavonoids, furanocoumarines, and cell wall components), existing in multiple isoforms [2, 3].

PAL plays an important role in catabolic processes of microorganisms, providing for utilization of L-phenylalanine as a sole source of carbon and nitrogen [4]. Among the microorganisms, the highest PAL activity is exhibited by yeasts, especially the red basidiomycetes of the Rhodotorula family [5]. Sporobolomyces roseus and Sporidiobolus pararoseus are also PAL-producing yeasts [6].

Therapeutic potential of PAL with respect to neoplasms was evaluated due to its selective activity to phenylalanine and amino acids that are consumed by mammalian cells from external sources. PAL was shown to inhibit neoplasm growth in vitro [7].

The enzyme is also of interest as a therapeutic agent for phenylketonuria treatment and may be used for both direct therapy of phenylketonuria and production of food products free of phenylalanine [8]. Besides the medical applications, the enzyme may be used in biotechnology for L-phenylalanine production from trans-cinnamic acid [9].

Considerable contribution to the development and assimilation of the technology of specialized food products was made by G.B. Gavrilov, N.B. Gavrilova, V.I. Ganina, N.I. Dunchenko, I.A. Evdokimov, V.I. Kruglik, K.S. Ladodo, L.A. Ostroumov, A.N. Petrov, V.O. Popov, G.Yu. Sazhinov, V.A. Tutel'yan, V.D. Kharitonov, I.S. Khamagaeva, and A.G. Khramtsov, and to the technology of the enzyme preparation of PAL, by V.I. Mushtaev, M. Jason Мас Donald, H. Orum, and O.F. Rasmussen.

The development of new and improvement of existing technologies of the PAL preparation production requires new, more intensive sources of its superexpression, which is impossible without studies on the specific features of its genetics in known producers. Only 26 sequences of genomes of microorganisms exhibiting PAL activity were found in the databases of genetic sequences (EMBL and GenBank). Therefore, the search for microorganisms exhibiting L-phenylalanine ammonia-lyase activity based on sequence analysis of their genomes is urgent.

The aim of the work was to screen and identify pigmented yeasts producing L-phenylalanine ammonia-lyase and describe their physiological and biochemical characteristics.

Список литературы

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13. Genosistematika - chto eto takoe? (Genosystematics: What is it?)

14. Babich, O.O., Prosekov, A.Yu., and Soldatova, L.S., Modeling of a continuous biotechnological process of L-phenylalanine ammonia-lyase production, Izvestiya vysshykh uchebnykh zavedenii. Seriya: Khimiya i khimicheskie tekhnologii (Higher Education Institution Herald. Chemistry and chemical technology series), 2013. V. 56 (4), P. 106-109.

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