CS201883B1 - Strain of microorganism saccharomyces cerevisiae ccy 21-4-63-e - Google Patents

Description

The invention relates to a strain of Saccharomyces cerevisiae which produces over 2% ergosterol in dry matter.

Saccharomyces yeasts are the most important ergosterol producers. The values reported in the older literature are relatively low. Proskurjakov, Popov and Oaipov (Biochimia 3: 397, 1938) found 0.31% in monofilament yeast, 0.55% in Torula utilis and 2.36% in dry matter for the Frohberg breed. Therefore, they consider brewer's yeast to be particularly rich in ergosterol, e.g. strain 776 had 2.5%, grape yeast had only 0.4 to 2.19% ergosterol in dry matter. Appleton and Kleber /Appl.Microbiol.3:249,1955/ Candida krusei demonstrated in 0.3% of C. parapsilosis even only 0, Q 6%, but _with the Saceharomyces cerevisiae to 9% of total sterols. Mitra and Garg / J.Inst.Brew. 64: 327.1958) determined 0.1-0.5% ergosterol in dry matter in the Dhar yeast, which they considered equivalent to other assays for Torula utilis. For more than 20 years, he has remained at a relative standstill in vitamin D, and has only risen in recent years. Jelizjan, Marojan and Arumjunian (Microbiology 44: 632,1975) examined the total sterol content of yeast and confirmed that the highest production is in carbohydrates, from 1.3 to 3.13% per biomass dry matter. Dikanskaja and Robyševa /Prikl.Biochem.Mikrobiol. 11: 21.1975 / found the ergosterol content of Candida, Torulopsis and Rhodotorula and found it to be between 0.4% and 0.6% on dry weight basis, and found only 2.4% for Saccharomyces carlsbergensis 4228. Latest Kosikov, Lyapuns,

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Rajevskaya and apol, / Microbiology 46: 66.1977 / report ergosterol production in yeast as follows: Odesskaja 14 baker's yeast produces 2.3%, other baker's yeast from 1.8 to 1.9%, medical races from 2.0 to 2 , 2% and brewer 4228 about 1.5% ergosterol in dry matter. From these cultures they isolated monosporococci, haploid and diploid, and a number of hybrids that produced from 1.3 to 2.3% ergosterol in dry matter.

The above-mentioned disadvantages of the previously known yeast strains are overcome by the invention, which is a new strain of the microorganism Saocharomyces cerevisiae deposited in Cs. yeast collection, Institute of Chemistry, Bratislava, Dubravska cesta, under number CCY 21-4-63-e.

The main advantage of the new strain is the increased productivity of ergosterol, which reaches up to 6% per biomass dry matter.

Strain CCY 21-4-63-e produces mild ellipsoidal cells with an average length-width ratio of 1.22, which has a surface / volume quotient of less than or equal to 1, in 20% to 1.2. It contains over 70% of cells with a P / 0 of less than 1. By repeated selection, the content of cells with a P / 0 of less than 1 can be further increased. The vast colonies of this strain are smooth and grow by about 1.5 mm in 100 hours. In a liquid environment, the trunk quickly sets off, on the surface there is no face, no skin, no wound or ring,

The CCY 21-4-63-e strain, after induction with acetite, sporulates well and can reach up to 80% of the sporulation elements. 2.5 of them are single spore, 18% double spore, 6Q% four spore. Disputes are round and smooth-walled. They need vitamins to grow in the environment. At 5 ° C it grows poorly (about 30% of the growth at the optimum temperature), at 42 ° C it grows very large, slightly (13% of the growth at the optimum temperature), the optimum growth temperature of the culture is 25-28 ° C. Ferments glucose, maltose, sucrose and raffinose. summer also sugars assimilates, does not assimilate potassium nitrate or lysine. It is an osmophilic yeast that can withstand up to 60% of saZ charozyme in the environment, 2% lactic acid stimulates its growth and 2% tartaric acid tolerates good. In the environment it can withstand 8% ethanol. It assimilates ethanol, glycerol, methanol and ethanediol.

The original strain 21-4-63 (the original culture labeled AgjMU-TDj ^ Trencin), which constituted only 10-20% of the cells with a P / 0 quotient of less than or equal to 1, was seeded in suspension, 10 'cells / ml per surface. the malt agar and a bead crystal was placed in the center of the dish on the agar surface. Under the influence of this mutagen, culturing was carried out in the dark at 28 ° C until aa isolated colonies of yeast appeared on the surface. By repeated sowing on the chord dishes, a mutant strain was produced that produced only cells with a surface coefficient (volume less than 1) at the ends of the exponential fes, ie at the time of maximum ergosterol production. This strain was designated CCY 21-4-63-e. ¹

The strain CCY 21-4-63-e can be used in a cultivation plant for the production of biomass, both in a classical way for yeast production and in a continuous process, using molasses or other waste sugar, or synthetic ethanol as a carbon source. The produced, centrifuged biomass is used to extract ergosterol and isolate it. The extracted biomass residues can be further processed to feed or used to recover other yeast biomass components / e.g. nucleic acids.

CCY 21-4-63-e strain was grown on Olson-Jensen artificial composition and induction molasses on a shaker and continuously. Ergosterol was determined after culture and lyophilization or directly using wet biomass. A 50-100 mg sample of 1. Lyophilized yeast was used for the assay, which was hydrolyzed in 40% potassium hydroxide for 1 hour at 0.196 MPa, shaken with diethyl ether for 2 hours on a shaker at 4 ° C. After separation of the ether layer, the aqueous layer was shaken again with ether and the combined ether extracts were washed well with distilled water. The pure ether extract was evaporated to dryness in a rotary evaporator. The residue was dissolved in 0.5 mL of chloroform and made up to 10 mL with ethanol, optionally diluted with chloroform: ether 5: 100. The ergosterol surge was checked with a registration spectrophotometer and the amount was calculated from the values at 282 nm.

Claims (1)

OBJECT OF THE INVENTION A strain of the microorganism Saccharomyces cerevisiae, CCY 21-4-63-e, which produces ergosterol.