CS209646B1 - Method of utilizing the sacharine acids - Google Patents

Description

1 ez refers to the method of utilization saccharinic acids. Saccharinic acids are acids of the summary formula ^ η ^ πθπ ’ which most often occur at the same time in the form of three structural.h types, both COOH and COOH 1 COOH 1 T H0-C-CH 2 OH j and HO-C-CH 1 1 COOH I 0 h 2 1 l CHOH 1 1 CH ₇ 1 1 CHOH | 1 CHOH I 1 CHOH 1 1 R 1 R 1 R isosaccharide type saccharin type metasaccharin type

wherein R is H or -CH _Z OH.

Saccharinic acids are unused and are found in various wastes from technologies in which hot alkaline agents are converted to polysaccharides, particularly in the alkaline pulp production processes where they are waste. The fermentative use of hydro acids is known and described for lactic acid, while the use of 2,4-dihydroxybutyric acid and other monocarboxylic hydroxyls containing five to six carbon atoms in its molecule is unknown.

This situation removes the invention, which is based on a method of utilizing saccharinic acids by using them as a raw material for the preparation of a culture medium in the form of a solution containing 30 to 40 parts by weight of a mixture of saccharinic acids;

containing phosphorus, potassium and magnesium, preferably in the form of 4 to 5 parts by weight of ammonium sulfate 0.5 to 0.7 parts by weight of 85% phosphoric acid -H3PO4, 0.5 to 0.8 parts by weight of potassium sulfate - K 2 SO 4 and 0.1 to 0.5 g of magnesium sulfate - heptahydrate - MgSO 4. 7H2O per 1000 parts by weight of a solution whose pH is adjusted to pH 3.5 to 11 with potassium hydroxide and used in this composition or treated with the addition of 30 to 50 parts by weight of mono- and disaccharides, preferably D-glucose or the addition of a lower aliphatic alcohol, preferably methyl alcohol or ethyl alcohol. To the culture medium thus prepared is added an inoculum of a culture of yeast or fungal microorganisms, preferably Caudida utilis, in the form of centrifuged yeast from their previous cultivation in a glucose solution of 0.2 to 0.5 Z containing an additional 2 parts by weight of ammonium sulfate. 3 parts by weight of phosphoric acid - 85%, 0.6 parts by weight of magnesium sulphate, adjusted to pH 5.5, which is carried out at 30 ° C, with constant shaking.

Culture medium and culture inoculum of centrifuged yeast or fungal microorganisms, added in an amount of 0,2 »» of their dry matter per 1 000 parts by mass of the culture medium, under intensive aeration or under anaerobic conditions at 25 to 60 ° C and shaking for 1 to 80 hours further by centrifuging the resulting biomass and drying yielded a crude protein-containing yeast solids using saccharinic acids other than glucometasaccharinic acid from the culture medium under aerobic conditions, and in an anaerobic culture method except metasaccharinic acid, which is preferably after concentration and treatment Extract the pH to 4.5 with ether and further utilize.

The advantages of said process are that as raw material for the preparation of the culture medium for microorganisms, waste from alkaline pulp production and from hot pulp refining can be used, from which cultures of microorganisms, especially yeast under successive conditions of aerobic or anaerobic fermentation capable of utilizing acids saccharin and isosaccharin. The fermentation process controlled by chromatography showed that the metasaccharinic acids are neutralized, which at the same time can be used for their isolation by an extraction method for further utilization.

The subject matter of the invention is illustrated by several examples without being limited thereto.

Example!

To 1 000 ml of a solution containing 31,5 g of a mixture of saccharinic acids composed of 12,5 g of glucoisosaccharinic acid, 4,5 g of glucoraetasaccharinic acid, 6,0 g of isosaccharinic acid, 7,0 g of saccharinic acid and 1,5 g of hydroxyraic acid are added 4.5 g ammonium sulphate,

0.6 ml of phosphoric acid (85%), 0.6 g of potassium sulphate and 0.3 g of magnesium sulfate heptahydrate and after mixing and dissolution, adjust the pH of the solution to 5.5 and add the inoculum of Caudida utilis in an amount of 0,2 Z calculated on the dry matter, in the form of centrifuged yeast from previously carried out on a 2% glucose solution, containing 2 g of ammonium sulphate, 0,3 ml of phosphoric acid (85 Z), 0,6 g of potassium sulphate and 0.1 g magnesium sulfate heptahydrate, adjusted to pH 5.5, which was transferred at 30 ° C on a shaker.

After fermentation, batch fermentation was performed in the fermentor for 10 hours at 32 ° C, blowing air at a rate of 1 liter / minute and 600 rpm on the turbine. Biomass is centrifuged and dried. 14.5 g of yeast dry matter were obtained, containing 56.2 Z of crude protein with digestibility coefficients of 87.5 Z.

Chromatographic examination revealed, with the exception of glucometasaccharinic acid, the full utilization of the other saccharinic acids.

Example 1 and d 2

To the mixture of saccharinic acids as in Example 1 and 50 g of D-glucose, 4.5 g of ammonium sulfate, 0.6 ml of phosphoric acid (85%), 0.6 g of potassium sulphate and 0.6 g of potassium sulfate are added. 3 g of magnesium sulfate heptahydrate and after stirring and dissolution the pH of the solution is adjusted to 5.5. Further procedure and fermentation conditions as in Example 1 yielded 43.5 g of yeast dry matter with a crude protein content of 59.1 Z.

Example 3

To do this, inorganic nutrients as in Example 2 were added to a mixture of saccharinic acids as in Example 1 and 25 ml of 96-th ethyl alcohol as described in Example 2. The further procedure and fermentation conditions as in Example 1 yielded 40.2 yeast dry matter.

Example 4

The procedure is to ferment to a mixture of saccharinic acids and D-glucose as in Example 2 and the addition of inorganic nutrients as in Example 2 and after adjusting the pH as in Example 2, but under anaerobic conditions during the culture. 80 hours. After isolation of the biomass and drying, the solution was concentrated to a volume of 50 ml and extracted after acidification to pH

4.5 ether. In this way. All types of saccharinic acids were utilized in the fermentation except metasaccharinic acid, which was obtained as described herein by 2.7 g.

Claims (4)

3 209646 EXAMPLE 2 To a mixture of saccharinic acids as in Example 1 and 50 g of D-glucose is added 4.5 g of ammonium sulfate, 0.6 ml of phosphoric acid / 85% by weight. 6 g of potassium sulphate and 0.3 g of magnesium sulphate-heptahydrate, and after mixing and dissolving, the pH of the solution is adjusted to 5.5. Further procedure and fermentation conditions as in Example 1 gave 43.5 g of yeast dry matter with a crude protein content of 59.1 Z. Example 3 Inorganic acid was added to a mixture of saccharinic acids as in Example 1 and 25 ml of 96% ethyl alcohol. nutrients as in Example 2. The following procedure and fermentation conditions as in Example 1 gave 40.2 yeast solids. EXAMPLE 4 A mixture of saccharinic acids and D-glucose as in Example 2 and addition of inorganic nutrients as in Example 2 and after pH adjustment as in Example 2 is fermented with the addition of a culture inoculum as in Example 1 but under anaerobic conditions for 80 hours. After biomass control and drying, the solution was concentrated to a volume of 50 mL and extracted after acidification to pH 4.5 with ether. In this way. all types of saccharinic acid metasaccharic acid were utilized in the fermentation, which was obtained in the manner described in 2.7 g. CLAIMS 1. A method of utilizing saccharinic acids consisting of a mixture of hydroxyls containing pal to six carbon atoms in the molecule structure of saccharine, isosaccharin and metasaccharin, characterized in that they are prepared from culture medium in the form of a solution further comprising the addition of inorganic nutrients, containing phosphorus, potassium and magnesium, preferably in the form of 4 to 5 parts by weight of ammonium sulfate, 0.5 to 0.7 parts by weight of phosphoric acid, 0.5 to 0.8 parts by weight of potassium sulfate and 0 to 10 parts by weight of ammonium sulfate. 1 to 0.5 parts by weight of magnesium sulfate heptahydrate per 1000 parts by weight of a culture medium solution adjusted to a pH of 3.5 to 11, preferably ammonium hydroxide, further mixed with an inoculum of a yeast or fungal microorganism, preferably Candidautilis, added in an amount of 0.2 of their dry matter per solution in the form of skimmed yeast from a a subsequent culture, in a 0.2 to 0.5 µl glucose solution, further comprising inorganic nutrients, preferably 2 parts by weight of ammonium sulfate, 0.3 parts by weight of phosphoric acid, 0.6 parts by weight of sulfate of potassium sulfate and 0.1 g of magnesium sulphate at pH 5.5, at 30 ° C with shaking and further aerating or under anaerobic conditions at a temperature of 25 to 60 ° C, with agitation and for 1 to 80 hours of accumulation of microorganisms . 2. Process according to claim 1, characterized in that from 1 to 10% of mono- or disaccharides, preferably D-glucose, are added to the saccharinic acid mixture. 3. Process according to claim 1, characterized in that from 1 to 10% of a lower aliphatic alcohol, preferably methanol or ethyl alcohol, is added to the saccharinic acid mixture. 4. Process according to claim 1, characterized in that the acid is isolated from the mixture after the fermentation under anaerobic conditions, after concentration and pH adjustment, by extraction. Scverography. n. p .. race 7. Most