SU994555A1 - Process for producing cellulolytic enzymes - Google Patents

Description

(54) METHOD FOR OBTAINING CELLULOLYTIC The invention relates to the microbiological industry, and specifically to a method for producing cellulolytic enzymes used in the process of pulping the pulp. A known method for the production of celluplastic enzymes involves the cultivation of the fungus Aspercg Pgus ierreus 17 P on cellulose-containing substrates that have been ground to 1-160 microns of rummers. Straw and guzapayu Clj are used as a substrate. A method of producing celluloaitic enzymes is known, involving the cultivation of D5regch Cus -terreus fungi in a nutrient medium containing an organic nitrogen source, mineral salts and sawdust as a carbon source. The strain AsperofiPEos terreusl7 P. C is used. The C and Cy activities are 6.2 and 2.4 mg of glucose / ml-h 2.

ENZYME. . The disadvantage of this method is the low activity of cellulolytic enzymes. The purpose of the invention is an increase in the activity of enzymes. This goal is achieved by the fact that according to the method of producing cellulolytic enzymes, involving the cultivation of fungi species (i | sf) evtri € Cu5 terreus on a nutrient medium containing an organic nitrogen source, mineral salts and sawdust as a carbon source, it was proposed as a producer use the strain flspertfieeosterreos VKM F 222O D, chop wood chips to a particle size of 0.2-1.0 mm. The strain sperofi BEos ierreos VKM JF-222P D isolated from forest soil Serpukhov district of the Moscow region. The strain deposited in the All-Union collection of microorganisms. The strain Asperj iRftus terreosBKM F 2 2 2OD has the following characteristic. 399 Cultural characteristics. Wednesday Capek. On day 9, when the colony reaches 7 cm in diameter; the edge of the knee is even, the surface is velvety, the central part is wrinkled-raised; the colony color is light brown, the opposite is brown, the pigment does not diffuse into the medium. Wort agar. The growth of the culture on the wort is different from the growth on the апapek medium by the presence of scant exudate, the center of the colony is raised at the injection site, the reverse side of the color of the medium. Morphological signs. Wednesday Capek. Conidial heads are long, column-compact ZO-5Och8O-150 yU conidiophores deviate from the substrate smooth, colorless 5-8 110140yU-} apical hemispheric bubble 14-18 / diameter, covered with sterigms 2/3 of its surface; double stigma, primary sterigma 2-2.5) .x7-8yu; , secondary | & 1E - 2-2,5.7,5-8,5Y; conidia elliptic, smooth 1.62, 2ua in diameter. Wort agar. On wort agar, there are differences in the sizes of some structures: conidia head 30-5O X80-20O / X-; conidiophore 5-8 11О180 / XJ diameter of apical bladder 10-16 d, primary sterigms 2-2.5 U х5.5-7d1-, secondary - 2-2.5 6-8 / х; diameter of conidia is 1.6-2.2 ui. . Tribe A5Rego | NoU5 terreus VKM P-222b D is able to grow in minerals with nitrogen sources using sawdust as the sole carbon source. Stasch does not need vitamins and other biologically active cotton. When organic additives are used, the activity of cellulases in the culture medium increases. Grinding of sawdust to the size of 0.1-1.0 mm has a stimulating effect on the synthesis of cellulases. Example. The mushroom spert ieeus ierreu VKM F-222O D is grown on a medium containing, g / l: birch sawdust crushed to a size of 0.2–1.0 mm; birch sawdust 20.0; NO N03 2.5; KH2PO4 2.0; Cc (Ce20.3; M 804-7H2O O, 3; yeast autolysate - 0.5; pH 5.5-6.0. Seeds are grown for 2 days at 40 ° C in flasks per 75O ml with a medium volume of 100 ml on a rocking chair at 180 rpm on a medium containing, g / l: lactose 54. lp, 0 | NUNOj 2.5; KH2P04 2 Oj Cbce / j 0.3; -MtfSO THjO 6.3; yeast autolysate; 0, 5; pH 5.5-6, O. The seed in the amount of 10% is added to the fermentation medium and incubated with a rocking chair at 180 rpm in 750 ml flasks with a volume of 1OO ml for 3 days. The culture medium is separated from the biomass of the fungus and the unused substrate, the centrifugation (5O OO rpm, 15 min), contains an active complex of cellulose C and Sc. - the activity of the culture liquid is, respectively, 1.5 and 40 mg of glucose / ml / h (activity is measured by the method of MapBase et e., Eiotechnoe. An. Bioeno; Samp., 6, 21-33, 1976; reducing sugars were determined by the method of Somoc vi MJ. BioB. Cliem. 195, 19-23, 1952). The endoglucanase and cellobiasis activity of the culture fluid is respectively 1.0 and 0.05 µmol / mlmin (activity measured by A.A. Kle method. owl and assoc. Bioorg. Chem., 6, No. 8, 1980, 1225-1242). PRI me R 2, the Mushroom spertfie u5 tert-eus VKM F -222OD is grown on a medium containing, g / l: birch sawdust crushed to a size of 0.2-1.0 mm, birch sawdust 2O; NaNO 2.5; KHgPOij 2.0; protein-vitamin concentrate 5.0. The inoculum is grown and farm 1cation is added to the medium as in Example 1. The culture medium separated after 2 days of cultivation contains a more active complex of cellulases: C and C activities are respectively 3.5 and 100 mg Glucose / ML1h, endoglucanase and cellobiasis activity 4.0 and 0, 1 μMOL / MLMIN. Thus, the described method will increase the yield of cellulases when using sawdust as a carbon source by 10-20 times in C-activity and 15-4O in Cj (- activity formulas of the invention lyuloliticheskih enzymes, comprising culturing fungi flsperc iCBus terreus species on a culture medium containing an organic nitrogen source, mineral salts and sawdust as a carbon source, otlichayuschiy5 .994555 6

with that. that, in order to increase the aqgi & - i. USSR author's certificate

Yuhugi enzymes, as producer No. 77222O, cl. C 12 N 9/42

STpppr r reus.2. P. Zeltin, P., Vitola, G / A. .. EDBKM-F-2220 D. In this case, tree crops I. M. Biosynthad cellulose smelting and particle size by the fungus pev ifeos ierr «« s

 17 P when cultivated in Bonjw

Sources of information, production conditions - In hectares, Ferprintye into account in the examination, Riga, -Zi Atne. .5-1оГ

Claims (1)

Claim A method for the production of cellulolytic enzymes, involving the cultivation of Asper <^ i8Kus F'eggee mushrooms in a nutrient medium containing an organic nitrogen source, mineral salts and wood sawdust as a carbon source, distinguishing 994555 with the fact that, in order to increase the activity of enzymes, producer use a strain of AspercpCCus terreus VKM F-222O D, while sawdust is crushed to a particle size of 0.2-1.0 mm