CS251532B1 - Microorganism strain Aspergilus tamarii CCM F-781 - Google Patents

Abstract

The solution concerns a new strain of Aspergillus tamarii CCM F-781, which produces icoadic acid in an increased yield. The essence of breeding and obtaining a new strain is that a suspension of Aspergillus tamarii spores with a concentration of 106/ml in dimethyl sulfoxide is treated with χ-rays at laboratory temperature for 25 to 35 minutes, while isolating the obtained mutants by standard selection methods.

Description

The present invention relates to a novel strain of Aspergillus tamarii which produces a koji acid in increased yield.

Aspe-rgillus tamarii has been described as a producer of kojic acid for a long time, but - without indicating the amount of feddic acid [Korzy-b-ski, T., Kofwezyk-Gi-n-dif, Z. Kurylowicz, W .: AntibiOtics , PWN - - Polish Scientific Publishers, Warszawa, (1967)].

According to a literature review, the Aspergillus tamarii strain is not used for industrial production of kojic acid. Catalogs of mushroom cultures (Cataloque of Cultures, Charles University, Prague 1976, No. 664 Aspergillus tamarii Kita; ATCC Cataloque of Fungi Yeasts, 16-th e-dition, 1984 issues 1005, 10 836, 12 699, 16 865, 20 054, 26 950, 26, 951 Aspergillus tani-arii Kit) disclose a species of Aspergillus tamarii without the ability to produce kojic acid. We do not have high-producing strains of fungi that are capable of producing kojic acid.

These disadvantages of the previously known strains are overcome by the invention which is a new strain of the microorganism Aspergillus tamarii mutartt VIII deposited in the Czechoslovak Milk Organism Collection of the University]. E. Purkyně in Brno, Class of Defenders of Peace 10, followed by CCM F-781. A new strain was obtained by treating a suspension of 10 ⁶ / ml Aspergillus tamarii spores in 10% dimethylsulfoxide for 25 to 35 minutes with χ-rays, spilling suspeinzia spores onto the sweetening agar and allowing it to sporulate at 25 ° C. up to 30 ° C, a spore suspension was prepared from the cultured culture, which was diluted 1: 1 000, spilled onto the sweetener -agar, where colonies of the microorganism culture were grown, which were evaluated by fermentation tests from an acid production level. and a strain of Aspergillus tamarii mutant VIII was selected therefrom.

The main advantage of the new quartz is the increased productivity, which reaches 42 kilograms of kojic acid per 100 kg of sugar. The original strain of Aspergillus tamarii had a maximum production of kojic acid of 15-18 kg per 10-0 kg of sugar. This strain on Sabouraud's pod in 3 days forms colonies with a diameter of 2.6 cm, the top side is white with strong sporulation in the middle of the colony. The underside is faintly orange. The spores are dark brown) of color 7 to 8 μΐη in diameter, ve -lukulum significant gufaté on average 44 to 48, αηι.

The new strain of Aspergillus tamarii mutant VIII grows well on Sa-b-ouraud agar. In 3 dini it creates colonies with a diameter of 5.3 cm without sporulation, the top side is white with high air mycelium, the bottom side is orange and smooth. It forms colonies with a diameter of 4 cm without sporulation on the sweet-agar in 3 days, the underside is radially wrinkled and orange-brown in color, the top is white. Sporulation on both soils occurs after 6 days of cultivation. The S-pores are dark brown in color, 7 to 8 ₍ µm in diameter, vesiculum -sightly rounded with a diameter of 56 to 6-0 / µm. Shelf life of spores is long-term when stored in activated carbon suspension in paraffin oil, or in lyo-freed s-taive for several years.

Example 1

The Aspergillus tam-aria suspension (10 ⁶ / ml in 10% dimethylsulfoxide) was treated with ⁶⁰ CO rays for 25-35 minutes. The spore suspension was then spilled onto the sweetening agar and allowed to stand. Spore in a thermostate at a temperature of 25 to 3-0 [deg.] C. A spore suspension in 0.9% onion sodium chloride is prepared from the prepared kit, diluted 1: 1000, and poured onto a slurry of agar, where in 3 min. After 4 to 4 days, microorganism culture colonies are grown, which are evaluated by fermentation assays for the production of kojic acid, and the strain Aspergillus tamarii mutwnt VIII is selected from them.

Example 2 (

A culture medium is prepared, using 100 g of glucose, 0.16 g of potassium dihydrogen phosphate, 0.07 g of sodium chloride, 0.02 g of manganese sulfate tetrahydrate, 1 g of urea, 1 g per liter. ammonium chloride, 0.01 g zinc sulfate heptahydrate and 0.01 g ferrous sulfate heptahydrate. A sterile culture medium of pH 5.0-6.5 in a 5 L fermentation tank is seeded with 10% v / v. 48 hour vegetative inoculum of Aspergillus tamarii muta nt-VIII, and cultured at 30 ° C, a peripheral speed of 3.1 m with ¹ and the aeration WM 3 for 5-6 days. During fermentation, 42 g / l of kojic acid is formed in the medium.

P r i k il a d- 3

The isolated strain has the property that its mycelium, after separation from the fermentation fluid, can be used several times for the fermentative preparation of kojic acid. A solution of 13-0 g of glucose in 1 L of 0.1 M phosphate buffer pH 6.0 is prepared. The sterile solution in a 5 L fermentation tank was mixed with the mycelium obtained by separation from the fermentation medium after the fermentation described in Example 2. The fermentation conditions were the same as in Example 2. 19 to 21 g / l of kojic acid are produced in the medium for 6 to 6 days of culture.

Aspergillus tamarii mutant VIII can be used in the fermentative production of kojic acid, which serves as a raw material for the chemical, food and pharmaceutical industries.

Claims (1)

251532 5 A Koji producing strain of Aspergillus tamarin CCM F-781 is present.