CS195049B1 - Method of producing beta-glucosidase enzyme - Google Patents

Description

AUGUSTIN JOZEF, PhMr., BRATISLAVA and POCKET WALL, Dr. , BRNO

Method of production of enzyme β-glucosidesis

The invention relates to the production of the [beta] -glucosidase enzyme (BC 3.2.1.21) by bacterial strains of Streptomyces epp., Cultured on suitable nutrient media,

For industrial production of β-glucosidase (β-D-glucoside glucohydrolase), the following microorganisms are most commonly used: Aspegillus oryzae, Merulius lachrymans (G. de Stevens in Method in Enzymology, Hive 1, SP Colowick and NO Kaplan, Acad Press). New.

York 1955, p. However, there are also other microorganisms which have not been known to date to have the ability to produce the β-glucosidase enzyme. Using the gel method (Author No. 183,169) of teething microorganisms for cellulase activity during the cultivation of microorganisms, it was believed that with 21 strains of the genus Streptomyces, 7 strains produced β-glucosidase.

The principle of the invention is that by culturing the following seven strains of the genus Streptomyces:

Streptomyces auréófaciens CCM 3229 W anulatus CCM 3158 levandulae CCM 3010 1 ' 'antibloticúe CCM 3159 M rutgensensis CCM 3196 griseocarneum CCM 3228

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Streptomycee diastatiene CCM 3167 individually or slightly mixed on cellulose-containing broth or its hydroxyethyl-, carboxyethyl-, hydroxypropyl derivatives at 25 ° C to 40 ° C and preferably pH 7.5 for 10 to 20 hours produces β-glucosidase enzyme which is obtained in the raw state by evaporation of the growth soil, in the partially reduced state, is obtained by precipitation with ammonium sulphate or an organic solvent, preferably ethanol and acetone.

For the purification of the crude β-glucosidase enzyme after palliation, e.g. ^ glucosidase adsorption of proteins to aluminum zmeei metahydroxyde and subsequently Release of adsorbed enzyme by Νβ 0θ2 to a solution of _2, as the G, Steven of the above references.

Examples of embodiment Example 1 _s

Streptomycee aureofaciens CCM 3239 is propagated on growth medium containing 2 g of gel (prepared by hydroxyethylcellulose blending according to Α.Ο.δ. 183 169) immersed in a half height in 20 ml of Yeast Nitrogen Base growth medium (0.6 g / 100 ml) and cultured is surface-treated for 3 days at a temperature of 25 to 40 ° C, during which the gel flows. The accumulated biomass of Streptomycee aureofaciens was used as a seed inoculum containing 20 g CORN-STEEP, 5 g hydroxyethylcellulose (pH 6.0) per liter of water for 25 hours. After 25 hours, the Streptomycee faciens biomass is centrifuged at 5,000 g for 10 minutes and the crude [beta] -glucosidase enzyme is obtained by evaporating the culture medium on a vacuum evaporator (total activity 55 IU, Specific activity 0.3 IU / mg protein).

Example 2

The procedure of Example 1, except that Streptomycee levandulae and Streptomycee rutgersensis was used for propagation.

The crude β2-glucosidase enzyme has a total activity of 45 10 and a specific activity of 0.35 IU / mg protein.

Example 3

The procedure of Example 1, except that the expanded culture of Streptomycee antibioticus was used as an inoculum on growth broth containing 20% gluten stillage and 7 g of water-soluble propylmethylcellulose per liter of soil (pH 6.0) for 24 hours.

Example 4

The procedure of Example 1b, except that the accumulated Streptomycee aureofaciens biomass was used as a seed inoculum containing 20 g CORN-STEEP and 1 g cellulose (pH 6.0) per liter of water for 15 hours. This yields a crude enzyme .gamma.-glucosidase with a total activity of 78 IU and a specific activity of 0.94 IU / mg protein.

The invention has utility in production. -glucosidases as one of the important enzymes in the recovery of various wood and agricultural wastes containing carbohydrate poles.

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P R E J) T E T OF THE INVENTION

Spéaob production® enzyme /? -glucosidesis by bacteria of the genus Streptomycee indicated

Claims (1)

P R E J) T E T OF THE INVENTION Spéaob production® enzyme /? -glucosidesis using bacteria of the genus Streptomycee characterized by the cultivation of strains of the genus Streptomycee: Streptomyces aureofaciena CCM 3239 M anulatus CCM 3158 W levandulae CCM 3010 11 tt antibioticua CCM 3159 H 11 griaeocarneum CCM 3228 you 11 rutgersenaia CCM 3169 tt diaatatiene CCM 3167 individually or mixed on a cellulose-containing culture medium or its hydroxyethyl-, carboxyethyl-, hydroxypropyl derivatives at 25 to 40 ° C and preferably pH 7.5 for 10 to 20 hours produces γ-glucosidase enzymes which aa awaits in the raw state by evaporation of the growth medium, in a partially attenuated state, by precipitation with ammonium sulphate or an organic solvent and preferably with acetone and ethanol.