CS263298B1 - Process for preparing fermentative extract and ergosterole - Google Patents

Abstract

It is a method of processing microorganisms, especially yeast, to obtain ergosterol and yeast extract. The advantage of the process is to achieve a higher yield of ergosterol (over 85%) without the need to saponify the lipid fraction while producing yeast extract. Neutral or acidic microorganism cells are heated vigorously from 0-25 ° C to 55-85 ° C and then cooled rapidly to 0-25 ° C after 5-20 seconds. Cooling must take place within one second before autolysis occurs after heating to 45 to 58 ° C. After separating off the solid particles from which ergosterol is extracted after disintegration, a yeast extract is obtained. The advantage of the process is that substantially less ergosterol losses occur in the process than in the prior art, and the other components of the cell are utilized so that especially the nitrogenous substances of the microbial cell are not impaired. The process is relatively simple, thus easy to implement. Waste is not difficult to dispose of.

Description

The invention relates to a process for the production of yeast extract and ergosterol.

The economics of ergosterol production from microorganisms depends on its concentration in the starting material. For this reason, efforts have been made to achieve the highest ergosterol concentration already at the fermentation stage. Attempts to influence the sterol content by culture conditions have led to an increase in the total sterol content, but mainly due to the ballad 24 (28) dehydroerosterol. The route of selection of a strain with a constitutively high content of ergosterol has proved far more advantageous. This compound contains up to 1.6% ergosterol. Further increase of ergosterol content by biological means is very difficult.

Methods of treatment of raw materials (eg yeast) by biochemical way, were intended to use some other cell parts before extract! ergosterol so that they do not deteriorate and at the same time ergosterol is concentrated. A method of obtaining cell walls of microorganisms for the production of ergosterol based on activation of lytiocyte cell enzymes by disintegration followed by autolysis for 10 hours is described (ref. AO 189 440). The separate cell walls then serve as the starting material for obtaining ergosterol. Similarly, AO 223 007 addresses the concentration of ergosterol in a small volume of ballast lipids following the process of AO 161 299. In both cases, it is the concentration of a portion of ergosterol to a certain fraction in the processing of yeast (microbial) cells. A disadvantage of the described and known processes is that only part of the ergosterol is concentrated. Its losses are offset by the acquisition of other products, but the loss of 30% and more of ergosterol is quite large.

This disadvantage is overcome by a process for the production of yeast extract and ergosterol, which is characterized in that the suspension of cells of microorganisms, preferably yeast, of neutral or acidic pH is subjected to a thermal shock by heating from 0 to 36 ° C to 55 to 85 ° C and After 5 to 20 s with cooling to 0 to 25 ° C in less than one second, the suspension is heated to 45 to 58 ° C, subjected to autolysis for 0.5 to 4 h and the yeast extract is separated from the solid particles.

A mixture of nonpolar solvent, toluene, petroleum ether, hexane or heptane with an addition of 5 to 20% by volume of an aliphatic alcohol is preferably used to extract ergosterol from the separated and after optional washing of the solid particles.

The proposed procedure hydrolyzes nucleic acids and proteins, leaving lipids, especially sterols, unchanged. Separation of the resulting low molecular weight substances leads to a loss of cell dry matter, a yeast extract and a dampening to a real concentration of ergosterol in the cells. The mass loss ranges from 20 to 30% of the original mass. The low molecular weight substances separated from the cell suspension can be used as a quality extract with suitable organoleptic properties after concentration and drying. The process according to the invention is very suitable for continuous production and thus for automation of the whole process. In addition, it is possible to disintegrate cells after concentrating ergosterol and extracting it with mixtures such as petroleum ether-ethanol (9: 1) without hydrolyzing (saponifying) the lipid components of the biomass. This simplifies the process quite a bit.

The following are examples which illustrate but do not limit the scope of the invention.

Example 1 wt. a suspension of yeast Saccharomyces oerevisiae in water at pH 4.5 and 21 ° C was pumped at a rate of 1300 ml / h through a copper exchanger designed to heat the suspension to 75 ° C for about 0.6 seconds and to wait at this temperature. s, including a conduit to the condenser which cooled the slurry to 20 ° C over 0.8 s. Thereafter, the suspension was fed into a flow-through vessel, where it reached a temperature of 53 ° C. The volume of this stirred thermostatic vessel was 3,000 ml, so the mean residence time was 2.3 h. After each 1 liter of suspension was collected (in a storage vessel, after flowing through the thermostatic vessel, solid particles (treated cells) were separated by centrifugation (10 min). The supernatant was separated and dried after concentration, the sediment (treated cells containing all ergosterol) was resuspended to a suspension of 6% by weight of dry matter, disintegrated and in a ratio of 1 part suspension, 2 parts ethanol and 18 parts petroleum ( 30 DEG-50 DEG C.), extracted for 1 hour with vigorous shaking, and the ergosterol was isolated from the organic phase The ergosterol content of the individual fractions during the procedure of Example 1 is shown in Table 1.

Table 1

Material Quantity of dry matter G Sterols G 24/28 dehydro -ergo- sterol G Ergo · ster < G suspension prior to extraction 130.0 2.10 0.40 1.70 tot. suspension after extraction 130.0 2.10 0.40 1.70 extracted cells (treated) 93.6 2.06 0.37 1.69 suspernatant extract after separation of the solid particles 37.1 0.01 0 0.01 organic phase after extraction of disintegrated processing cells 1.85 0.40 1.45

The total ergosterol gain is more than 85% of the original amount.

Example 2

In the same manner as in Example 1, 10 wt. suspension of Candida utilis yeasts grown on sulphite extracts. Hexane was used instead of petroleum ether for extraction. The sterol content of the individual fractions is shown in Table 2

Table 2

Material Dry mass Δ5,7- sterols 24/28 dehydro- ergosterol Ergosterol G G G G

orig. suspension 130.0 0.60 0.13 0.47 suspension after extraction 130.0 0.58 0.11 0.47 extracted cells treated 98.5 0.58 0.11 0.47 extract (supernatant after separation of solid particles) 31.5 0 0 0 organic phase (hexane) after extraction of the disintegrated treated yeast 0.51 0.10 0.41 The total ergosterol gain is 87 % of the original quantity.

The loss of ergosterol in the treatment method according to the invention is up to 20%, which achieves more advantageous parameters than, for example, according to the method according to AO 189 440. Moreover, it is possible to extract the homogenate directly after disintegration. and there is no need to saponify.

Claims (2)

Process for the production of yeast extract and ergosterol, characterized in that a suspension of cells of preferably micro-organisms, preferably yeast, of neutral or acidic pH is subjected to heat shock, heating from 0 to 36 ° C to 55 to 85 ° C and after 5 to 20 with cooling to 0 to 25 ° C in less than one second, after which the suspension is heated to 45 to 58 ° C, subjected to autolysis for 0.5 to 4 h and the yeast extract is separated from the solid particles which are extracted. 2. Process according to claim 1, characterized in that a mixture of a non-polar solvent, toluene, petroleum ether, hexane or heptane with an addition of 5 to 20% by volume of aliphatic alcohol is preferably used for extracting ergosterol from the separated and after optionally washing the disintegrated solid particles.