SU732381A1 - Actinomyces albogriseolus 28-3(vkm a-588) strain as glucoseisomerase producent - Google Patents

Description

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This invention relates to the microbiological industry and relates to the production of enzymes.

A known strain of Streptomyces albus 5 ATCC 21132 - producing glucose isomerase 1.

The disadvantage of this strain is its low productivity and enzymatic activity (

GLI / ml protein).

Having increased ability to biosynthesis of intracellular glucose isomerase and high productivity, the strain Actinomyces albogri-. seolus 28-3 (VKM A-588) is derived from soil samples. Stored in the collection of the MITIC under the number 28.-: and in the All-Union Collection of Microorganisms under the number VKM A-588. „

Cultural-merphological signs.

Sporosy spirals, spirals stretched from two to six curls. The spores are oval with a smooth and warty surface.

Synthetic medium CP-1 with glucose. Aerial mycelium scanty, chalky white (DZ). Substrate mycelium fawn (04); no pigment.

Waxman Wednesday. Aerial mycelium is very poor, whitish (D1). Substrate mycelium dark cream (On); no pigment. Wednesday Gauze No. 1. Aerial mycelium is poorly developed, Le E foreign gray (A4), Substrate mycelium dark cream (06); no pigment.

Soeva: Wednesday. Aerial mycelium scanty, mealy, fawn (04); substrate mycelium golden yellow (M7); no pigment.

Oats on Wednesday. Aerial mycelium is moderately developed, dark ashen (B4). Substrate mycelium dark cream (06); there is no pygmy.

Wednesday Gause 2. Aerial mycelium is developed by separate islets, whitish (D1). Mycelium pale honey (PZ); no pigment.

Corn environment 2. Aerial mycelium is absent, the stroke is highly developed, folded. Sacrificial mycelium of dark cream (06); no pigment.

The coloring of the mycelium is given according to the Bonartsev scale.

Physiological and biochemical properties.

Aerofil, the optimum temperature for growing is about 30 C.

Culture does not form a melanoid pigment.

Starch hydrolyzes (hydrolysis zone 3 mm).

Gelatinu liquefies.

Milk does not coagulate and does not peptonize.

Attitude to carbohydrates. It absorbs D-glucose, sucrose, D-xylose, i-i. Inositol, D-mannitol, D-fructose, rhamnose. Does not assimilate raffinose.

An example. Strain Actinomyces albogriseolus 28-3 (VKM A-588) is etched for seven days on beveled xylose-starch agar in test tubes. The composition of the medium,%:

Soluble potatoes1, starch starch 1.0

D-xylose 1.0

Peptone

Yeast Extract 0.5

HgSOif 0.1

2 - 6HgO

CoCl. 0.024 Agar CDifco) 2, 0 7.0, str medium before sterilization

relieve at 0.5 ati for 30 min.

The resulting spore material seeded flasks containing 35 ml of the main nutrient medium of the following composition,%;

1.0

D-xylose 1.0

Peptone 1.0

Yeast Extract 0.3

K.jHPOy

MgSOy. 7Н „00,1

Agar CDifco) 0.1

pH before sterilization 5.5, sterilization at 0, 5 for 30 minutes

Flasks with inoculated nutrient medium are placed on circular rocking machines (200 rpm). Cultivation temperature is 30 ° C. The inoculum grown during 4 hours (5 ml) is used for inoculation of 50 ml of fresh basic nutrient medium. The cultivation of the producer in the second stage is carried out for 24 hours, as in the first stage.

The obtained culture fluid is centrifuged for 15 min at 5,000 seb / mn; the separated biomass is washed twice with distilled water, destroyed and centrifuged for 25 minutes at 8000 rpm. The resulting supernatant is used as an enzyme preparation.

The determination of the activity of the isomerization reaction is carried out in a mixture of the following composition, ml: 0.2 M phosphate buffer (pH 7.0) 2.52.5

1 M D-glucose1,0

O, 1 M MgsO; - 7H2OO, 5

Enzyme preparation 1.0 The total volume of the mixture was adjusted to 10 ml with distilled water. Reaction

It is carried out in a water bath for 60 minutes, and an equal volume of 0.5 M HC is stopped by adding. The amount of D-fructose formed is determined by the cysteine-carbazole method. Per unit of glucose activity, an amount of enzyme is taken that results in the formation of 1 ml of D-fructose for 1 hour at 70 ° C. The specific activity of the glucose isomerase in the resulting preparation is 8-9.5 GLI / mg protein. The dry weight of biomass ranges from 10 to 14 kg / ml of medium.

Claims (2)

Invention Formula Strain Actinomyces albogriseolus 2b-3 (VKM A-588) - producing glucose isomers. Stored in the collection of ICTD at number 28-3 and in the All-Union Collection of Microorganisms under the number VKM A-588. Cultural and Formal Techniques. Sporosum spiral, spiral stretched from two to six turns. Spores are oval with a smooth and warty surface. Synthetic medium CP-1 with glucose. Aerial mycelium scanty melovo-white. Substrate mycelium fawn, no pigment. Waxman Wednesday. The air mycelium is very poor, whitish. Substrate mycelium warm cream; no pigment. Wednesday Gause number 1. Aerial mycelium is poorly developed, mouse gray. Substrate mycelium dark cream; no pigment. Soyeva Wednesday. Aerial mycelium scanty, mealy, fawn. Substrate mycelium golden yellow; no pigment. Oats on Wednesday. Aerial mycelium is moderately developed, dark ashen. Substrate mycelium dark cream; no pigment. Wednesday Gause number 2. Aerial mycelium is developed by separate islets, whitish. Mycelium pale honey; no pigment. Corn environment No. 2, Aerial mycelium is absent, the stroke is highly developed, folded. Mysticium substrate dark cream, no pigment. Physiological and biochemical properties Aerofil, optimum temperature for growing near. Culture does not form a melanoid pigment. Starch hydrolyzes. Gelatinu liquefies. Milk does not coagulate and does not peptone. Attitude to carbohydrates. Absorbs D glucose, D-xylose, sucrose, 1-inoit, D-mannitol; D-fructose, rhamnoe. Not assimilated raffinose. Sources of information taken into account in the examination i. U.S. Patent 3,616,221, Cl. 195-31, published; 1971.