SU1703689A1 - Strain of fungus penicillium canescens producent of fructodofuranosidase - Google Patents

Abstract

This invention relates to the microbiological industry, in particular to the preparation of a new producer strain / 3-fructofuranosidase. used in the food industry and agriculture. The purpose of the invention is to obtain a strain / 3-fructofuranosidase with a higher activity. The strain Penicillium canescens VKPM F-418 obtained by treating the original strain with ultraviolet, deposited in the Central Museum of industrial microorganisms VniIgenetika. During deep cultivation on a medium containing soy flour and mineral salts, the strain produces 5-fructofuranosidase with an activity of 80 U / ml (72 hours). The activity of the purified enzyme 240 u / ml In addition to / 3-fructofuranosidase, the producer synthesizes / 20-galactosidase 20 units / ml of bg-galactosidase - 3.5 units / ml. xylanase - 5 units / ml (Ls

Description

The invention relates to the enzyme industry, in particular to the preparation of a new strain, the producer of the enzyme ft-fructofuranosidase. used in the food industry and agriculture.

The purpose of the invention is to obtain a strain with a higher activity of / -fructofuranosidase.

To obtain a highly active strain, the strain Penicillium canescens Sopp was used. 20171 (with an activity of 7.0 units / ml), which is treated with ultraviolet rays at a dose of 200 J / m.

The strain deposited in the Central museum of industrial microorganisms VNIIgenetika. collection number VKPM F-418.

Morphological signs. The strain is toxic, fast-growing, characterized by moderate spore-carrying. Conidia formation begins one day.

later than in the original culture. Conidia 2.2-2.5 microns, ovoid, then sharog.idnye. rough, chained in a column, but then more or less splayed during aging. Colom .- and on agar wort, agar medium of čapek, agar medium with sucrose, fluffy, consisting of dense basefelt felting and loose wreath of the airway mycelium with poorly expressed radial subcutaneously with abundant spraying. superior to the original crop by yield of spores by 10–15%. Colonies with a growing margin of 2-3 mm width.

VI

ABOUT

with about

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about

white to pale gray, then smoky gray with aging. Conidiophores deviate from the substrate to 350–500 x 3–3.5 µm or as short sprigs of air spores with variously arranged splayed 1.2–3 whorled structures with swells usually around 10–20 x 2 µm, carrying sterigms 8 x . 2.3 microns with a clear conidiophen neck, narrow, but relatively short.

Cultural features. On agarized artificial media (wort agar, potato-glucose agar, potato agar) and synthetic agar media (Czapek, Saburo, Rolen-Tom, and agar medium with sucrose), the strain grows well.

Linear growth of strain colonies on agarized media at the bottom of the day: wort agar medium 5th day 35-48 mm: 10th day - up to 85 mm; Wednesday of čapek - 5th day 30 mm: 10th day 40 mm; Lactose agar medium - 5th day 28-40 mm; 10th day to 70 mm.

For cuts under conditions of solid phase fermentation, the strain grows well.

Attitude to carbon nutrition. Good by itself ... assimilates monosaccharides: glucose, arabinose, sugar alcohols: sorbitol, lures, dulcite, from disaccharides: lactose, sucrose, maltose, as well as starch and inulin.

Attitude towards nitrogen nutrition: assimilates nitrate nitrogen, preferably ammonium. From organic nitrogen sources, it assimilates peptone, urea, yeast autosate, corn extract.

Attitude to oxygen: obligate aerobic.

Relation to temperature: mesophilic, optimal temperature for growth and biosynthetic activity is 26-27 ° C.

Relation to the acidity of the nutrient medium: grows over a wide range of pH from 2.0-9.0.

For breeding the strain, a modified Czapek medium with sucrose of the following composition was used,%: sucrose 3.0; SIZE 0.2; KH2P04 0.1, MgS04 0.05; KCI 0.05; corn extract 0.05, agar 2, initial pH 5.8-6.0.

The nutrient medium for fermentation is the medium of the following composition,%: Soya flour 3.0; Na2HP04 12H20 15; (NH4) 2S04 0.2; KCI 0.05, MgS04 0.015, initial pH 4.1-4.2.

An inspection carried out over a period of four years has established that when stored in

Seeding the level of enzyme activity of Penicillium canescens AME-85 is not reduced.

The producer forms a protein rich biomass (up to 30% by weight of dry biomass).

The strain Penicillium canescens AME-85 support on agar medium with sucrose, wt.%: Sucrose 3.0; MSMD 0.2, KH2P04 0.1; KCI 0.1; MgS04 0.05, corn extract 0.05; agar 2. Baseline pH 6.0.

Deep cultivation of the fungus is carried out on a liquid nutrient medium of the following composition, wt.%: Soybean flour 3.0:

Na2HP04 12H20 1.5; (0.2; KCl 0.05; MgSO4 0.015; initial pH 4.2.

The cultivation is carried out in 750 ml flasks with a thermostatic rocking chair, which makes 220 rpm (level

aeration 1.4 02 / lh) at 30 ° C for 72 hours. The volume of the nutrient medium in the flasks is 100 ml. Sowing the nutrient medium Edy produced 2% suspension of conidia of the surface culture of the fungus grown in

test tubes on beveled sucrose agar for 12 days at 28 ° C.

The biomass is separated by filtration and the activity of the / -fructofuranosidase is determined in the filtrate using the Samoji-Nelson method. The reaction mixture consisting of

0.7 ml of substrate. 0.5 ml 0.2 M acetate

pH 4.5 buffer, 0.3 ml enzyme solution,

incubated at 50 ° C for 15 min,

then 1 ml of Samoji's reagent is added and

boil in a water bath for 20 minutes. After cooling, 1 ml of Nelson's reagent is added, made up to 5 ml with water and photometrized at 590 nm. The activity of α-3-fructofuranosidase is 80 units / ml (known

57 units / ml).

In the filtrate, besides the activity of (9-fructofuranosidase), a-galactosidase (3.5 units / ml), / -galactosidase (20 units / ml) and xylanase (5 units / ml) are contained.

The filtrate is separated and the i-fructofuranosidase is isolated as a separate fraction in which the activity is determined.

The activity of the purified α-fructofuranosidase is 240 units / ml.

Claims (1)

Claims of the invention The strain of the fungus Penicillium canescens VKPM F-418 is a producer of / 3-fructofuranosidase.