CS201882B1 - Microorganism strain Saccharomyces cerevisiae CCY 21-4-67-e - Google Patents

Abstract

The invention relates to the strain Saccharomyces cerevisiae, which produces over 2% ergosterol in dry matter. Yeasts of the genus Saccharomyces are the most important producers of ergosterol. The values reported in older literature are relatively low. For more than 20 years, research in the field of vitamin D has remained relatively quiet, and has only increased in recent years. The disadvantages of the literature of previously known yeast strains are eliminated by the invention, which is a previously unknown strain of the microorganism Saccharomyces cerevisiae, deposited in the Czech Collection of Yeasts, Chemical Institute of the Slovak Academy of Sciences, Bratislava, Dúbravská cesta, under number 21-4-67-e. The main advantage of the new strain is the increased productivity of ergosterol, which reaches values of up to 6% per dry matter of biomass. Strain CCY 21-4-67-e produces ellipsoidal cells with an average aspect ratio of 1.30, whose surface/volume ratio is less than or equal to 1, in 20% to 1.10. It contains over 80% of cells with P/0 less than 1. By repeated selection, the content of cells with P/0 less than 1 can be increased somewhat. The huge colonies of this strain are smooth and oily, shiny, growing by 1.57 mm in 100 hours. In a liquid medium, the strain sediments, does not form a skin, rind or ring on the surface.

Description

Microorganism strain Saccharomyces cerevisiae CCY 21-4-67-e

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. For more than 20 years, he has remained at a relative standstill in vitamin D, and has only risen in recent years.

Disadvantages of the yeast strains known from the literature are eliminated by the invention, which is a hitherto unknown strain of the microorganism Saccharomyces cerevisiae deposited in Czechoslovakia. yeast collection, Institute of Chemistry, Bratislava, Dubravska cesta, under number 21-4-67-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 21-4-67-e produces ellipsoidal cells with an average length-width ratio of 1.30 whose surface / volume quotient is less than or equal to 1 in 20% to 1.10. It contains over 80% 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 huge colonies of this strain are smooth and greasy shiny, increasing by 1.57 mm in 100 hours. In a liquid environment, the strain sedimentes and does not form a skin, shell or ring on the surface.

201 882

The CCY 21-4-67-e strain, after induction with acetate, sporulates well and can reach up to 62% of sporulation elements. Of these, 2% is single-spore asc, 16% is two-spore, 82% is three-spore. Disputes are smooth and gul’ate. To grow it needs vitamins in the environment. It grows weakly at 5 ° C and at 42 ° C, the optimum temperature being 25 to 28 ° C. Ferments glucose, galactose, maltose, sucrose and raffinose. It also assimilates, does not assimilate potassium nitrate or lysine. It is an 8-cell yeast that tolerates up to 60% sucrose in the environment. 2% lactic acid does not affect growth, 2% tartaric acid inhibits growth by 50%. In an environment of 8% ethanol. It assimilates ethanol, ethanediol and methanol.

The original CCY strain 21-4-67 (the original culture designated TK 3 Libon) was inoculated in suspension! 10 < 6 > cells / ml on the surface of the wort agar and in the center of the plate on the surface of the agar was placed a crystal of the bead. Cultivation was carried out in a thorn at 28 ° C until isolated yeast colonies appeared on the surface. By repeated sowing on the dishes, a mutant strain was produced that produced only cells with a surface / volume coefficient of less than 1 (towards the ends of the exponential phase, i.e. at the time of maximum ergosterol production). The strain produced an increased amount of ergosterol of from 3.5 to 6% per bionic dry matter. This strain was designated as CCY 21-4-67-e.

The CCY 21-4-67-e aa strain can be used industrially for biomass cultivation, both by classical yeast production and in a continuous process, using molasses or other waste sugar, or synthetic ethanol as a carbon source. Unproduced, centrifuged biomass ea is used to extract ergosterol and isolate it. The extracted biomass aa residues can be further processed to feed or can be used to recover other yeast biomass components / e.g. nucleic acids.

The CCY 21-4-67-e strain was grown on an Olaen-Jensen artificial composition and inductively on a shaker and continuously. Ergosterol was determined after culture and lyophilization or using wet biomass. A 50 to 100 mg sample of lyophilized yeast was used for the assay, which was hydrolyzed with 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 with distilled water. Pure ether extract aa was evaporated in 0.5 ml chloroform and made up to 10 ml with ethanol, optionally diluted with chloroform: ether 5: 100. The ergosterol demonstration was checked with a recording apectophotometer and the amount was calculated from the values at 282 nm.

Claims (1)

2 201882 The strain CCY 21-4-67-e is well-induced by acetate induction and can reach up to 62% of sporulation elements. Of these, 2% are single-spore quizzes, 16% double spores, 82% triple and quadruple spores. The disputes are smooth and guléaté. It needs vitamins in the environment to grow. Slightly rising at 5 ° C and at 42 ° C, optimum temperature is 25 to 28 ° C. Glucose, galactose, maltose, sucrose and rarinose ferment. They also assimilate these sugars, do not assimilate potassium lysine. It is an osmophilic yeast that can carry up to 60% of sucrose in the environment. 2% lactic acid does not affect growth, 2% tartaric acid inhibits growth to 50%. It contains 8% ethanol. Assimilates ethanol, ethanediol and methanol. The initial strain CCY 21-4-67 (a "culture" labeled "TK 3 Liben") was inoculated in a 10 'cell suspension to the surface of the wort agar and crystals deposited on the surface of the agarubol. Cultivation took place in a thorn at 28 ° C until a rare yeast colony appeared on the surface. By repeatedly spreading on the plates, a mutant strain was produced which produced only cells with a surface / volume coefficient of less than 1 / end of the exponential phase, i.e. at the time of maximum ergosterol production. The strain produced an increased amount of ergosterol from 3.5 to 6% on dry matter bioni ^ ey. This strain was designated CCY 21-4-67-e. The strain CCY 21-4-67-e aa can be used industrially for cultivation in order to obtain a biomass, both in the classical way for yeast production and in a continuous way, using molasses or other waste sugar feedstock, or synthetic ethanol and carbon. Non-produced, centrifuged biomass is used to extract ergosterol and isolate it. The extracted biomass residues can be further processed to feed or molluscs used to obtain other yeast biomass components (e.g. nucleic acids. Strain CCY 21-4-67-e was grown on Olsen-Jensena pdde artificial composition and on induction on a shaker and continuously. Ergosterol was determined after cultivation and lyophilization or using wet biomass. A 50-100 mg sample of lyophilized yeast was used for the assay, which was hydrolyzed with 40% potassium hydroxide for 1 hour at 0.196 MPa, washed with diethyl ether for 2 hours on a shaker at 4 ° C. After separation of the ether layer, the aqueous layer was washed once more with ether and the combined ether extracts were washed with distilled water. The pure ether extract was evaporated in 0.5 mL of chloroform and filled with ethanol to 10 mL, optionally adjusted with 5: 100 chloroform: ether, the ergostero proof was checked with a registration spectrophotometer, and the amount was calculated to be 282 nm. SUMMARY OF THE INVENTION The strain of Saccharomyces cerevisiae CCY 21-4-67-e producing ergosterol. Price 2.40 Kčs TZ 6/70