CS195021B1 - Method of producing beta-glucan glucanohydrolase with bacterium streptomyces spp - Google Patents

Description

BACKGROUND OF THE INVENTION The present invention relates to the production of [beta] -glucan glucanohydrolase (EC 3.2.1.4) enzyme by bacterial strains of streptomyces spp. cultivated on a suitable nutrient pod.

The following microorganisms are most commonly used for industrial production of β-glucan glucanohydrolase α and :richoderma viride, Trichoderma coningia, Neurosposa crassa and others.

However, there are also other microorganisms which have not previously been known to have the ability to produce β-glucan glucanohydrolase. Using the gel method / Cs. AO No. 183 169 / testing of microorganisms for cellulase activity during culture of microorganisms, we found that it produced β-glucan glucanohydrolase from 21 strains of the genus Streptomyces 14.

None of these strains of the genus Streptomyces is known in the literature as a producer of cellulase enzymes, although many strains of the genus Streptomyces are used for the production of antibiotics, where the accumulated bacterial biomass contains cellulase enzymes. The principle of the invention is that by culturing the following 14 strains of the genus Streptomyces:

Streptomyces flavovirens CCM 3243 vinaceus CCM 3005 ' « distatochromogenes subst. luteus CCM 3013 H griseus CCM 406 «T griseoluteus CCM 3242

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Streptomyces californiaua CCM 3016 H ·· gelaticus CCM 3177 « olivaceus. CCM 3181 H fradiae CCM 3174 « venezuelae CCM 3198 flaveolus CCM 3171 h albogriseolus CCM 3227 tt viridochromogenes CCM 3201 H scabies CCM 3197

individually or mixtures thereof on a culture medium containing cellulose or its derivatives (hydroxyethyl, carboxymethyl, hydroxypropyl) at 37 ° C and preferably pH 7.5 for 10 to 20 hours produces the β-glucan glucanohydrolase (E.C) enzyme. 3.2.1.4/, which is obtained in the raw state by evaporation of the growth pod, or partially tempered by precipitation with ammonium sulfate (60% saturation), ethanol or acetone (1: 1). All mentioned microorganisms are stored in Cs. collection of microorganisms of J.E.Purkyně University in Brno, Class of Defenders of Peace 10.

Examples of design

Example 1

Streptomyces gelaticus CCM 3177 aa multiplies in a growth fall containing 2g of gel prepared by crosslinking / J. Zemek and L. Kununi: Cs. AO No. 183 169 (immersed in half-height in 20 ml of Yeast Nitrogen Base growth medium (0.6 g / 100 ml)) and cultured superficially for 3 days at 37 ° C, when the gel is liquefied, Streptomyces gelaticus accumulated biomass used as the inoculum of the falls containing 20 g of CQRN-STEEP, 5 g of hydroxyethylcellulose (pH 7.8) per liter of fall for 15 hours. After 15 hours, the crude Streptomyces gelaticus is centrifuged at 5000 g for 10 minutes and the crude enzyme is obtained by evaporating the culture flask to a vacuum evaporator (total activity 40 IU, specific activity 0.26 IU / mg protein).

Example 2

As described in Example 1, and thereby disintegrating, a mixture of Streptomyces olivaceus and Streptomyces fradiae was used for propagation. An auric enzyme is obtained with a total activity of 38 IU and a specific activity of 0.32 IU / 1 mg protein.

Example 3

As described in Example 1, except that a multiplied culture of Streptomyces gelaticus was used as a growth fall inoculum containing 20% gluten stillage and 7 g of voco-potassium carboxymethylcellulose per liter of pH 7.5 for 16 hours. Quicker processing as in Example 1. Auric enzyme with a total activity of 29 IU and a specific activity of 0.24 IU / 1 mg protein is also obtained.

The invention has utility in the production of β-glucan glucanohydrolysis as one of the important polysaccharide hydrolase enzymes, which has application possibilities both in basic research and in the practice of utilizing polysaccharide-containing woody and poinohoapodic wastes.

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Claims (1)

. OBJECT OF THE INVENTION Sposdb production of β -glucan glucanohydrolysis using Streptomyces app. characterized by culturing the following 14 strains of the genus Streptomyces: Streptomyceá flavovirens CCM 3243 vinaceus CCM 3004 M distatochromogenes aubst.luteus CCM 3013 Streptomyces griseus CCM 406 M griseoluteus CCM 3242 w californicus CCM 3016 M gelaticus CCM 3177 N olivaceus CCM 3181 ti fradiae CCM 3174 H venezuelae CCM 3198 M flaveolus CCM 3171 11 albogriseolus CCM 3227 ti viridochromogenes CCM 3201 11 scabies CCM 3197 individually or a mixture thereof on cellulose-containing nutrient medium or its hydroxypropyl-, carboxymethyl-, hydroxyethyl derivatives at 37 ° C and preferably pH 7.5 for 10 to 20 hours, produces the enzyme γ-glucan glucanohydrolase, which is obtained in crude in a quenched state by precipitation with ammonium sulphate or an organic solvent, preferably ethanol and acetone.