CS205298B1 - Manufacturing method of alpha-amylase - Google Patents

Description

The invention relates to a process for the production of α 1 -amylase.

Currently, the enzyme alpha-amylase is obtained by isolation from the production medium of Bacillus subtilis and Streptomyces aureofaciens. Alpha-amylase production is also described in Saccharomycopsis capsularis / H. Ebert, Folia microbiologica 11: 422-438, 1966], Saccharomycopsis fibuliger and Saccharomycopsis fibuliger var. var. hordei / US patent no

297 547 /. The disadvantage of the production of alpha-amylase by bacteria is a worse separation of biomass from the culture medium and a lower saccharification capacity of the obtained alpha-amylase.

The above-mentioned disadvantages eliminate the method of alpha-amylase production on the liquid nutrient medium contained. sources of assimilatable carbon and nitrogen and mineral nutrient salts, the essence of which is that strains Sc hwannicot with occidentalis CCY 47-1-1 and CCY 47 - 1-4 # Sc hwa nn i omyc es alluvius CCY 47-2 -1 are cultivated singly or in a mixture thereof on a culture medium containing 0.1 to 3 wt. % soluble starch at 25-35 ° C, pH 5-7, for 48-60 hours, wherein alpha-amylase is obtained by evaporation of the growth stage, optionally by ammonium sulfate precipitation or affinity purification.

The advantages of said method of alpha-amylase production are the simple separation of biomass from the culture medium, the health of the biomass over fungal fermentations and the fact that the reaction amylolytic product of Schwannníomyces alpha-amylase is rich in maltose. The alpha-amylase activity obtained is 44 U and the specific activity is 2.9 U / rag of biomass, which is equal to the production capacity of fungal producers.

Example 1

Triangular profiles of 1.5 cm were cut from the prepared amylose gel, placed in a Petri dish and poured into Yeast nitrogen base, Difco; 0.657 g per 100 ml of distilled water. Gels are immersed to about half their height. After repeated sterilization three times at 100 ° C in steam, a culture of Schwanniomyces occidentalie CCY 47-1-1 is inoculated on the gel surface and cultured at 30 ° C. The liquid culture is microscopically checked for biological purity and used as an inoculum on a pod containing 1 Z corn steep, 0.5 Z radical sulphate and 0.2 Z potassium dihydrogen phosphate and 12 dissolved starch (per 0.2 L) in an Erlenmeyer flask (1). PH adjusted to 6.2. After 48 hours of cultivation! at 28 ° C, the Schwanniomyces occidentalis biomass is removed by centrifugation and the resulting supernatant containing α 1 -a-araylase is processed. The crude enzyme is obtained by concentration of the culture medium in a vacuum evaporator or by precipitation with ammonium sulphate at 58% saturation of ammonium sulphate. The purified enzyme can be obtained using an affinity method.

An advantage of preparing a cross-linked amylose inoculum is that said polysaccharide is calmable in a large population with high alpha-amylase production that is relative to a lesser production population in the pre-vivation.

Example 2

The procedure is as in Example 1, except that the strain Schwanniomyces alluvius CCY 47-2-1 is cultured on a nutrient broth containing 10 g per 1 liter of soluble starch, at 35 ° C, pH = 5, for 55 hours.

Example 3

The procedure is as in Example 1, except that Schwanniomyces occidentalis CCY 47-1-4 is cultured on a nutrient broth containing 30 g per 1 of soluble starch, at 25 ° C, pH 7, for 60 hours.

Claims (1)

OBJECT OF THE INVENTION Process for the production of alpha-amylase on a liquid nutrient medium containing sources and imitated carbon and nitrogen and mineral salts, characterized in that the strains Schwanniomyces occidentalis CCY 47-1-1 and CCY 47-1-4 and Schwanniomyces alluvius are cultivated individually or in a mixture thereof on a nutrient medium containing 0.1 to 3% by weight of dissolved starch at 25 to 35 ° C, pH 5 to 7, for 48 to 60 hours, wherein α 1 -α-araase is obtained by evaporation of the growth medium, ammonium sulfate precipitation or affinity purification.