SU1033543A1 - Method for preparing mannite - Google Patents

Abstract

A method for producing mannitol by cultivating its producer on a nutrient medium containing carbohydrates as a carbon source, yeast extract and necessary mineral salts, followed by concentration and crystallization of the target product, characterized in that, in order to increase mannitol yield and reduce fermentation time, the quality producer use the strain Rep1s11liuni decumbens BIMG-138.

Description

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The invention relates to the microbiological industry, namely to a method for producing mannitol used in the food, medical and chemical industries, in perfumery, as well as in the preparation of surfactants, drying oils, resins and varnishes.

A known method for producing mannitol by cultivating the yeast of the genus Torulopsis on media containing glucose or maltose, or glycerin, mineral salts, yeast extract at 28-30 seconds, concentrating under vacuum and crystallization.

The disadvantages of the inlet method are the duration of the fermentation process and the use of expensive medium.

The closest technical solution to the invention is a method for producing mannitol, which consists in growing the fungus Aspergillus Candidas in a nutrient medium containing carbohydrates, such as glucose as a carbon source, yeast extract and mineral salts, concentrating and crystallizing mannnt 2.

The most significant drawbacks of this method are the low yield of mannitol (up to 50%, the duration of maintaining fermentation (813 days, and the use of an expensive component of the medium, glucose, as a carbon source.

The aim of the invention is to increase the yield of mannitol and reduce the fermentation time.

The goal is achieved by the fact that according to the method of producing mannitol, which involves cultivating its producer on a nutrient medium containing carbohydrates as a carbon source, yeast extract and necessary mineral salts, followed by concentration and crystallization of the target product, the producer of RepiciIlium decumbens BIMG-138 is used as a producer ,

The strain of the fungus Penicillium decumbens BIMG-138 obtained by multiple passages of the parent form of Penicillium decumbens and the selection of the most productive colonies for mannitol. The strain is deposited in the Museum of cultures of industrial microorganisms of the Institute of VNIigenetics, collection number, SCHPM F175. The strain is stored in the Museum of Cultures of the Institute of Microbiology of the Academy of Sciences of the Byelorussian SSR at 4-5 ° C on vol-agar

The strain of the fungus Penicillium decumbens, BIMG-138 has the following characteristics.

Morphological signs. Conidiophores 50-10-X 2-2.5 microns smooth or finely rough, often formed in the marginal zone from stolonovidnymi

hyphae along the substrate. Sterigms in compact bundles of 12-15, 7-9 x X 2-2.5 microns. Conidia are elliptical to spherical, 2-2.5 microns sometimes up to 3 microns, greenish in mass, loose columns up to 100 microns in length, often falling away in culture.

Cultural features. Wort-aga seven-day colonies with a diameter of 3 cm velvety, light grayish-greenish-white. Aerial mycelium is more developed in the center of the colony and has a cotton structure. On the reverse side of the colony is unstained with the surrounding colorless agar. Spore formation begins at 4 days of growth.

Czapek’s medium is dense — seven-day colonies with a diameter of 2.8 cm. The colonies are velvety, greenish-white. Grow limited. Aerial mycelium is developed in the center of the colony. The colonies on the reverse side are pale yellow in color with the surrounding colorless agar. Conidiophena zone in the center of the colony is greenish. The formation of a dispute occurs on day 7. When sowing on a dense nutrient medium, the growth appears for 2 days, and by 6-7 days the colonies become velvety with abundant sporulation. In a liquid nutrient medium, the growth appears after 1 day and by 3-5 days a large amount of fungal biomass fungus consisting of dense pellets accumulates, and the content of mannitol in the culture fluid reaches 13-15 g / l of medium.

Molasses-agar-seven-day colonies with a diameter of 3.5 cm. The colonies are velvety, greenish-white, green in the center. Aerial mycelium is highly developed over the entire surface of the colony, as a result of which it has a cotton-like structure. On the seventh day, weak spore formation is observed.

Biochemical signs. Gelatinu dilutes very slowly, superficially. Peptonized milk, restores nitrates, starch hydrolyzes.

Relation to carbon sources; assimilates rhamnoe, arabinose, xylose, fructose, mannoe, glucose, sucrose, maltose, glycerin, dulcit. It grows poorly on galactose and practically does not utilize lactose.

Molasses is used as a carbon source - a waste of sugar production, or sucrose, or glucose.

Example. The seed is grown on a rocking chair at 27-30 ° C with a stirring speed of 200 rocking

/ min for 2 days in 300 ml Erlenmeyer flasks containing 50 ml of medium of the following composition, g / l: KHjPO 1) 0.5

.HCl 0.5-, FeS04 0.01; yeast extract 5, glucose 30. The pH of the medium is 6.0. The resulting inoculum in the amount of 10 vol.% In a sterile environment. After the same composition and. Grow two days. The main environment of the composition ,, g / l: N3N03 2; one ; 0.5 KS1 100; FeO4 0, EXHAUST gas, - yeast extract 17.5) glucose 30, pH 5.0, sown with prepared 4-day foreign exchange in an amount of 10 o6.%. Cultivation was carried out on a rocking chair in 300 ml Erlenmeyer flasks containing 50 ml of medium; at 27–30 s and the mixing speed is 200 swings / min. After 96-120 hours, the fungus mycelium is filtered. The filtrate is deionized on ion exchange resins, concentrated under vacuum to obtain a 25-30% solution of mannitol, and allowed to crystallize in the cold. The precipitated crystals are separated by filtration or centrifugation. The yield is 48-50% of the carbon source used. Example 2: The main sterile medium containing g / l: NaNOj 2; KH2P04 1 “, MgSO —THjO KCl. FeSOji 0.01) yeast extract 12.5; glucose 30; PJ 5,0 - sown with prepared 4-day seed in the amount of 10 vol.%. Cultivation was carried out on a shaker with a stirring speed of 200 swings / min at 27-3Q ° C for 4-6 sous in 300 ml Erlenmeyer flasks containing 100 ml of medium. The yield of mannitol is 57.5 from the carbon source used. PRI me R 3. Prepare the medium soy-. Tava is analogous to Example 1, but instead of glucose, 30 gUl of sucrose is added. The conditions for the cultivation and extraction of mannitol are the same. The yield of mannitol is 54.5% of the carbon source used. Example4. The culture conditions and medium are the same as in Example 2, but 30 g / l sucrose is used instead of glucose. The yield of mannitol is the carbon source used ... li p i me r 5. The cultivation conditions and medium composition are similar to example 1, but instead of goats. 50 g / l molasses is added. The yield of mannitol is 48.0% of the carbon source used. Note 6. The composition of the medium and the culture conditions are similar to those of Example 2, but 50 g / l molasses is added instead of glucose. The yield of mannitol is 64.0% of the carbon source used. The use of the proposed method provides an increase in the yield of mannitol to 64.0% of the used carbon source against 50% of the known, reduction of cultivation time to 4-5 days against 8-13 hours of the known, the use of cheap nutrient medium .

Claims (1)

METHOD FOR PRODUCING MANNITITE by cultivating its producer on a nutrient medium containing carbohydrate as a carbon source, yeast extract and necessary mineral salts, followed by concentration and crystallization of the target product, characterized in that, in order to increase mannitol yield and reduce fermentation time, in As producer use the strain 'Penicillium decumbens BIMG-138. 00 00 SP oo>