CS259288B1 - A method of complex fractionation of baker's yeast - Google Patents

Abstract

The solution concerns a method of selective utilization of baker's yeast. The essence of the solution lies in the fact that the supernatant of the yeast autolysate is precipitated with ethanol at a temperature of 2 to 5 °C in a volume ratio of 0.9 to 2:1, the residue is dried after removal of ethanol, the lipid fraction is extracted from the autolysate sediment with a 20-fold excess of the ethanol-hexane extraction mixture in a volume ratio of 1:2, which is further fractionated with a 5-fold excess of acetone, while the acetone extract, after saponification with a 15% potassium hydroxide solution, is further extracted with hexane for 1 h at 80 °C, concentrated and after adding acetone to a final concentration of 33% by weight. in hexane crystallizes at a temperature of -5 °C, while the delipidized residue remaining after the extraction of lipids from the autolysate sediment is dried after dilution with water to an 8 to 10% suspension.

Description

250203

The present invention relates to a method of complex fractionation of baker's yeast.

Yeast biomes, including bakery dies, are the source of many valuable substances that have so far been isolated mostly, with the remaining part of the yeast cell content unused.

Proteins are obtained from yeast most often by alkaline extraction, respectively. by various modifications of this method after disintegration of cells (Gooney, CL, Rha, C., Tan-nenbaum, SR, Adv. Food, Res. 26, 1-52,1980 j. The main obstacle to the use of such microbial proteins There is a high level of nucleic acids in human life, and there are several ways to reduce the nucleic acid content of biomasses and hence in the isolated protein product. (Rut, M., Stros, F., Hladecek, P., Kvasny prumysl '24, 58-66; Czechoslovak invention no.188,575), the effect of chemical agents on intact cells (Damodaran, S., Kinsella, JE, Biotechnol. Bioeng. 25, 761-770, 1983; U.S. Patents 3,615,654 and 3,775,393), using thermal shock (Deimann, W., Fabri-ni HD, J. Exptl. Med., 23, 883-897, 1979; U.S. Patent No. 2,081,172), enzymatic cleavage of nucleic acids by nuclease both endogenous and exogenous (Rott, D., Fencl, Z., Food Industry, 30, 531-3). 1979; U.S. Patent No. 3,867,255).

Both the ergosterol and the total sterols are most commonly isolated by a method which includes saponification steps prior to extraction into the organic solvent. The unsaponifiable steroidal fraction is obtained by extraction with a non-polar solvent, most often hexane, acetone, or extraction benzene (No. 189,440).

Invertase is one of the industry's most widely used enzymes. A traditional method for the release of yeast invertase from a cell is the auolysis induced by elevated temperature or by chemical means. In particular, precipitation with organic solvents, most commonly ethanol, polyethylene glycol, diethyl ether (Neumann, P., Lampen, JO, Biochemistry 6, 468-75, 1965; Bacon, S., Methods Enzymol. 1, 251-262, 1955; No. 101971).

The yeast cell wall polysaccharides are most often isolated by partial alcaline hydrolysis, e.g., glucans and mannans (Manners, DJ, Masson, AJ, Patron, JC, J. Gen. Microbiol. 80, 411-417, 1974), or cell wall of yeast is enzymatically disrupted by trypsin, insoluble polysaccharides are freed from dissolved glycides and protein degradation products by swelling (Pillemer, L., Blum, L., Lepow, IH, Wurz, L., Told, EW, J) Exptl. Med 103, 13-13 (1956). With this procedure, zymosan (a mixture of glucans and mannans), which is a preparation with immuno-chemical properties, can be populated. Preparation procedure for CSAO 185 063).

The disadvantage of these procedures is the inefficient use of the feedstock. This disadvantage is eliminated by the method of the invention, wherein the yeast autolysate supernatant is precipitated with ethanol at a temperature of 2 to 5 ° C in a volume ratio of 0.9 to 2: 1, the residue is dried after removal of the ethanol , from the autolytic sediment, the lipid fraction is extracted with a 20-fold excess of the ethanol-hexane extraction mixture in a 1: 2 volume ratio, which is further fractionated with a 5-fold excess acetone, wherein the acetone extract is saponified with a 15% solution of potassium hydroxide for 1 hour. h at 80: C, further extracted with hexane, concentrated and, after addition of acetone, the resulting mixture. 33 wt. in hexane, crystallizes at -5 ° C, where the delipidized residue, which remained after the extraction of lipids from the sapo autolysate sediment, with water to 8-10% suspension, is dried. The yeast autolysate is separated mainly by the protein supernatant and sedimented by non-permeable polysaccharides, lipid fractions, protein residues and nucleic acids, which are bound to cell walls during autolysis. The safinalized supernatant is spray dried by drying in a protein concentrate with a high crude protein content, or the fraction is used to recrypt the preparation by invertase organic solvent, or the appropriate volume ratio of the organic solvent is added to the obtained supernatant at reduced temperature, the precipitate is converted by centrifugation and dried as protein concentrate. After complete dilution with water, the complete sediment fraction is dried in a spray dryer as a biosorbent for the winery or the lipid fraction is extracted quantitatively by a mixture of organic solvents from the sediment. The solvent is evaporated from the lipid extract, the residue is washed with excess acetone while lipids are not added to the solution. After extraction, the insoluble fraction of polar lipids remains in acetone, which is suitable as an emulsifier, for example for the preparation of instant starch. The sterol concentrate can be obtained by hexane extraction of the saponifiable fraction from the total lipid extract of the sample fraction. It is suitable for the production of viral Dz. Thus, the advantage of complex fractionation of the invention is the selective use of baker's yeast. Another advantage of this is that the entire process is a waste-free technology that is low on material and technical security. The principle of complexity leads to some compromises in the processes compared to the solo isolation of individual components, but the yields and quality of the individual products are satisfactory enough. The method of complex fractionation according to the invention can be over-modified by the stages of the products that are at the center of interest, in particular by changing the initiation factors of autolysis, influencing its course, or changing the process of isolation and product finishing. From one batch of baker's yeast, a protein concentrate of prelud nutrition containing 40-80% protein can be obtained, a coarse preparation of invertase with activity of 6–23 ukat / g. Dry matter suitable for confectionery applications, ergosterol with 80% purity pharmaceutical production of vitamin Da the fraction contains 30% of phospholipids, used as an emulsifier for the preparation of instant dross, and a product containing 70% of polysaccharides from which immunoactivates can be obtained, mananes for chemical applications, respectively. hydrolyzate in biotechnological nutrients. Example 1

Disintegration 2,550 ml of a 10% yeast bakery yeast cell suspension is initiated by the addition of 425 ml of fresh yeast autolysate, 26 g of NaCl and 128 getanol. Autolysis is carried out with stirring at 50 ° C and is terminated after 5 hours. The autolysate containing 153 g of crude protein is immediately dried or centrifuged to give 2400 ml of supernatant, releasing 38% protein from total protein in yeast cells. In the sediment, unchanged lipid and polysaccharide levels are retained compared to the tin cells. 1a) The supernatant is dried in a spray dryer to obtain a consistency product of white color with a protein concentration of 67% by weight, or 1b) 0.9 to 2 times the volume is added to the supernatant 96% ethanol, while ethanol supernatant is cooled to 4 ° C. 4.5 g of precipitate with activity of the convertase 9 are obtained per gram of dry matter. The convertase preparation is suitable for use in foodstuffs, or the same procedure as in Example 1b, but after separation of the invertase precipitate by centrifugation, the fugate is dried in a spray concentrator of the dryer. 1d) The sediment is diluted with water and dried in a spray dryer. The obtained product has favorable adsorption properties, comparable to commercial preparations of foreign companies used as biosorbents in the vegetation to stimulate the fermentation of grape seeds, or le) the sediment is saponified by a 3-mass amount of ethanolic KOH solution (1 mol / l) for 3 hours under poor cooler. After completion of the alkaline hydrolysis and cooling of the contents, distillation is added in an amount corresponding to 1/3 of the total volume of hydrolyzed sediment. The mixture is mixed and extracted 3 times with n-hexane in a 1: 1 volume ratio. the solvent is evaporated and the asterol concentrate is blown through a stream of inert gas. By this process, up to 80% of the total yeaststerols of 50% purity can be obtained. The preparation can be used directly in agriculture for de-ratification purposes, or after further purification and conversion to vitamin D2 in pharmacy.

If the ethanol is extracted from the sediment with a 20-fold excess of extraction extract, ethanol-hexane in a volume ratio of 1: 2 is concentrated. The extract is concentrated and added to it. 500 ml of ace-tone. Concentrated acetone extract solidified with 200 mL of 15% methanol / CH 2 Cl 2 / CO 2 for 1 hour and extracted with bexane. The hexane extract was evaporated to a final volume of 100 ml. Acetone is added to this volume and the solution is cooled to CG5 CG. This procedure yielded 3.26 g of a yellow-needle needle preparation containing 1.99 gsterols, representing 69.8% of the contents of intestinal yeast contents. The resulting stearic fraction can be used for the preparation of the viral Dz for pharmaceutical applications or

The Igj lipid fraction is extracted from the residue with a 20-fold excess of ethanol-hexane extraction mixture in a volume ratio of 1: 2. The solvent is evaporated. The residue is extracted 4-5 times with 5-fold excess acetone. After extraction, the non-soluble fraction of polar lipids (about 3.5 g), which contains 30% phospholipids, crosses in acetone. This fraction can be used in the preparation of the instant yeast. Acetone extracts of saspoja, the process of further processing being analogous to Example 1e. 1h) Delipidized cell walls obtained by extraction of the lipid fraction according to If and lg procedures are filtered off, homogenized and dried with a spray dryer after water control. This gives a fine powdery product which contains 68-73% polysaccharides, suitable for the isolation of pure glucans for immunopharmaceutical purposes. Example 2

The same procedure as in Example 1, the autolysis was terminated after 24 hours. More than 87% of the total protein content of the yeast cells is released into the supernatant in the flowcholyolysis. 2a) The same procedure as in Example 1a, the protein product contains 76% of white-wines. 2b) The same procedure as in Example 1b, however, is obtained. 15 g what is it

Claims (1)

259288 7 Invertase preparation with an activity of 6.8 ukat per gram of dry matter. Example 3 The same procedure as in Example 1, however, using a volume of 10 ° / o suspension of baker's yeast - 1700 ml. Autolysis is initiated by the addition of 170 ml of fresh yeast auto-lysate, 2 g of sodium chloride and 17 g of ethanol. The autolysis is carried out at 50 ° C with stirring and terminated after 10 hours. Autolyte contains 90 g of crude protein. Centrifugation yielded 1300 ml of supernatant at 596 g of sediment, releasing 30% of the total protein content of the yeast cells into the supernatant. 3a j The same procedure as in Example 1a, however, the protein product contains 80% crude protein. B) The same procedure as in Example 1b, by precipitation, 1.7 g of a preparation with invertase activity of 23.5 µg / g of dry matter is obtained. OBJECTS A method of complex fractionation of baker's yeast characterized in that the superfine of the yeast autolysate is precipitated with ethanol at 2 to 5 ° C in a volume ratio of 0.9 to 2: 1, the residue is dried after removal of the ethanol, the autolysate is extracted from the sediment the lipid fraction with a 20-fold excess of ethanol-hexane extraction mixture 1: 2, which is further fractionated with a 5-fold excess of acetone, the acetone extract being washed with 15% hydroxide solution The extract was extracted further with hexane for 1 h at 80 ° C, concentrated and, after addition of acetone, the resulting concentration was 33%. in hexane, crystallized at -5 ° C, while the delipidized residue, which remained after the lipid was extracted from the autolysate sediment, was dried by diluting with water to 8-10%. Severografia, n. P. Plant 7, Most Price 2,40 Kcs