Mikrobiol. Z. 2018; 80(2):28-43. Russian.
β-Mannanase Activity of Yeasts Isolated in Antarctic
Borzova N.V., Gladka G.V., Varbanets L.D., Tashyrev A.B.
Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine
Psychrophilic and psychrotolerant microorganisms of the Antarctic are known as producers of enzymes with unique properties, in particular, resistance to extreme environmental conditions. The aim of the work was to establish the species afliation of three antarctic yeast cultures and to study their glycosidase activity. Methods. To study the enzymatic activity of yeast, synthetic and natural substrates were used: p-nitrophenyl-glycosides, guar galactomannan, MC cellulose, soluble starch. Isolation of genomic DNA was derived from cell suspensions. Fragment of the 26S rRNA gene in strains U5 and U8 were PCR amplifed using primers NL1 and NL4, in strain S181 fragment of the 18S rRNA gene PCR amplifed using primers NS3 and NS6. The resulting sequences of genes of yeast isolates compared with those of microorganisms deposited in the database of GenBank, using the BLASTN software package. Phylogenetic analysis was performed with programs ClustalX 2.1, Mega 6.06 (Neighbor-Joining). Results. In the culture supernatant of strains exo-α-rhamnosidase and endo-β-mannanase activity were detected. It was shown that high of β-mannanase activity and productivity of two psychrotolerant strains occurred on the third day, and one on the second day of cultivation, and correlated with the synthesis of the protein. Growing cultures at 15 °C provided higher rates of extracellular β-mannanase activity. The optimal sources of nitrogen were chloride and ammonium sulfate, and carbon − guar galactomannan and rhamnose. As a result of phylogenetic analysis of nucleotide sequences of 26S rRNA or 18S rRNA genes, it was established that the strains studied belong to the species Cryptococcus victoriae and Cryptococcus terricola. Conclusions. The high β-mannanase activity of antarctic strains of C. victoriae and C. terricola and the prospect of their use as producers of mannan-degrading enzymes are shown for the frst time.
Keywords: psychrotolerant yeast, Cryptococcus victoriae, Cryptococcus terricola, β-mannanase.
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