SU745933A1 - Trichoderma viride 44 mold fungus strain as cellulolytic enzyme producent - Google Patents

Description

(54) STRAIN mold GRIVA Tgishoderma viride 44 The invention relates to an enzyme industry mikrobkologicheskoy industry and relates to a novel strain producing, extracellular cellulases are used at ensilage roughage, in direct duction alcohol, as well as for the preparation of sugar solutions of the waste food and wood industry . Various strains of mold fungi are known, including the Triehoderma viride species, the enzymes of which are used for the hydrolysis of cellulose-containing plant substrates Cl3. A disadvantage of the known producers of cellulases is the low activity of the C1 enzyme. The closest in technical essence to the present invention is the strain Trichoderma viride, All-Union Research Institute of Genetics and Selection of Industrial Microorganisms. This strain is characterized by the following features. When sowing a prick on Rowlen-Thom's agar medium with the addition of corn extract, on the 7th day of growth, colonies are produced PRODUCT CELLSOLYTIC ENZYMES of the strain, which occupies the entire surface of the cup. Aerial mycelium is white, powdered; the periphery of the colony is not lowered; Conidia in the form of dark green pads are densely gathered in the center of the colony. The medium is not colored. The culture is capable of utilizing glucose, glycerin, starch, cellulose. The milk is coagulated, followed by peptonization, the final pH is 7.8. Gelatin dilutes slowly. The optimal temperature for the growth of the strain on agar and liquid nutrient media is 28-30 C. Breathing is aerobic. Cellulolytic enzymes are produced. The activity of the Cj-enzyme is 5.5 U / ml 23. The purpose of the invention is to obtain the HOBord strain, which is ensured by the increased activity of cellulose. The strain of the fungus Trichoderma viride 44 was obtained from the museum strain Trichoderma viride; VNIIGenetics (registration) by three stages of selection using heat treatment and chemical mutagens.

745933

 , L1Tamm Trichoderma viride 44 - a producer of cellulolytic enzymes is cultivated in deep conditions on a liquid nutrient medium ;;;; ;;;;;; /; ; / 7.,., „. . .,. , 1, .. L ..,., L

The Cj-enzyme cellulase activity is determined using filter paper as the substrate using the Schsmodyi-Nelson method,

The activity of cellulase C-enzyme in the culture fluid is 22-25 units / ml for 6 days of growth.

Morphological signs.

Mycelium septirovan, conidiophores are formed in the form of lateral branches of the hyphae in 30-32 h growth / poorly differentiated, sometimes branched. Conidia oval, size 2.5-2.8 X 3.5-3.7 microns lemon-yellow color. .

Cultural features.

On wort aratie at 5 days of growth at 30, the diameter of colonies is 3–3.8 cm. Aerial mycelium is white, concentric colonies, center is raised, do not grow into agar, conidia are lemon-yellow in color, yellow pigment is released on Wednesday.

On potato-dextrose agar for 5 days, the diameter of the colonies is 3.0-3.2 cm, the growth pattern is as on medium with wort agar. On the environment of čapek with starch for 5 days, the diameter of the colonies is 2.53, 0 cm, the growth is weak, the mycelium is in the form of a light bloom, the conidia are yellow, located throughout the entire colony, the medium is colored yellow.

On Chapek-Doksa medium with sucrose for 5 days, the diameter of the colonies is 3.03, 2 cm. Growth pattern is similar to that of čapek with starch. The medium is not pigmented.

On the Gatchinson medium for 5 days, white loose mycelium is formed on the surface of the paper, the medium and the strip are blue. SYSKSKY SIAeifiii B yymbno-yellow color.

On slices of potatoes are potato growth, fluffy mycelium, white, horse-colored lemon yellow, a slice of potatoes is colored yellow.

On the Rowlen-Tom medium on day 7 of growth, the diameter of the colony is 5.0-5.5 cm, the mycelium is white, powdered, and conidia of yellowish color cover the entire surface of the colony. The medium is not colored.

Physiological and biochemical signs.

Assimilates glucose, galactose, lactose, maltose, xylose, mannose, sucrose, glycerin. Cellulose hydrolyzes to glucose. Starch hydrolyzes. Gelatin does not dilute.

Milk does not coagulate. Uses ammonium, nitrate and protein nitrogen forms.

Strain intensively produces

cellulase with a growth on medium-sized beet pulp. A maximum of akoplenin cellulases in culture liquid is 144 h of growth.

When cultivating a strain of Trichoderma viride-44 in a fermentor with a capacity of 3x) l activity of Ci-enzyme

for 4 days is 17-18 u / ml

for 6 days 22-25 units / ml.

Thus, the proposed strain allows to increase the activity of culture cultures from 5.5 to 20-25 U / ml, which provides an increased yield of the drug.

The strain is characterized by abundant sporulation, which is of great importance for production in view of the reduction in the amount of seed.

The economic effect is 1333 rubles / m of culture fluid. Formula of the invention

The strain of mold fungus Trichoderraa viride 44 is a producer of cellulolytic enzymes. It is stored in the Museum of Cultures of the All-Union Scientific Research Institute of Genetics of Industrial Microorganisms. Registration CMPM F-105.

Morphological signs. Mycelium septate. Conidiophores are poorly differentiated, forming by 30-32 h of growth as lateral branches of hyphae, sometimes branched. Conidia are oval, 2.52, 8x3.5-3.7 microns in size, contain one lemon-yellow color.

Cultural features.

On wort agar on the 5.th day of growth at 30, the diameter of the colonies is 3.5-3.8 cm. White aerial mycelium, colonies

concentric, do not grow into the agar, the center is raised, the conidia are lemon yellow, a yellow pigment is released into the medium.

On Rowlen-Thom medium, skugorous extract on day 7 of growth, the diameter of colonies is 5.0–5.5 cm, mycelium is pale yellow, powdered. Conidia are yellow, the medium is not colored.

0 On potato-dextrose agar for 5 days of growth, the diameter of the colonies is 3.0-3.2 cm, the growth pattern is as on medium with wort agar.

On the environment of čapek with starch on

f 5 days of growth, the diameter of colonies is 2.53, 0 cm, the growth is weak, the mycelium is in the form of a light one is put on, the conidia are yellow, the medium is colored yellow.

. On the environment of čapek-Doksa with sucrose for 5 days of growth the diameter of the colonies

0 3.0-3.2 cm, the nature of growth on the environment of čapek with starch. The medium is not pigmented.

On the middle of Getchinson, on the 5th day of growth, white loose mycelium forms on the surface of the paper, medium-5,74593 and a strip of paper are colored lemon-yellow. Growth is good on slices of potatoes, fluffy white mycelium, conidni of lemon-yellow color, slice. potatoes are yellow in color.5 Physiological and biochemical characteristics. Assimilates glucose, galactose, maltose, xylose, mannose, sucrose, glycerin, lactose, Cellulose is hydrolyzed to glucose, .Q starch hydrolyzes, gelatin is not. thins, milk does not coagulate. Uses ammonium, nitrate and protein forms of nitrogen. The strain intensively produces cellulase when growing on a medium with 36 :: vellum pulp. The maximum accumulation of cellulose is 144 hours. The activity of the C | -enzyme for 4 days is 17-18 units / mp, on the 6. day 22-25 units / mp, Sources of information truncated into account during the examination 1. USSR author's certificate 399522, cl. C 12 D 13/10, 1973. 2.Kalun Nts K.A., Vaganova M.S., Shus G.V., Chugunova T.D. and Grachev Yu, P, Optimization of the composition of the nutrient medium for the biosynthesis of cellulase by the fungus Trichoderma. - Applied; biochemistry and microbiology, 1976, t. XII, vol. 2, s. 269-272 (prototype).

Claims (1)

20 claims The mold fungus Trichoderma viride 44, a producer of cellulolytic enzymes, is stored in the 25th Museum of Culture of the All-Union Scientific Research Institute of Genetics of Industrial Microorganisms. Registration number TsMPM F-105. Morphological signs. jQ Mycelium septic. Conidiophores are weakly differentiated, they form at 30-32 h of growth in the form of lateral branches of hyphae, sometimes branched. Oval conidia, size 2.5 £ 5 2.8x3.5-3.7 microns, contain one core of lemon yellow. Cultural ^ signs. On wort agar, on the 5th day of growth at 30 *, the diameter of the colonies is 3.5-3.8 cm. The aerial mycelium is white, the colonies are 40 concentric, do not grow into the agar, the center is raised, the conidia is lemon yellow / yellow pigment is released onto the medium. On a Rowlen-Tom medium with maize extract, on the 7th day of growth, the diameter of the colonies was 5.0-5.5 cm, the mycelium was pale yellow, powdery. Conidia are yellow, the medium is not colored. 50 On potato-dextrose agar, on the 5th day of growth, the diameter of the colonies is 3.0-3.2 cm, the growth pattern is similar to that on a medium with wort agar. On Chapek’s medium with starch, on the 5th day of growth, the diameter of the colonies is 2.53.0 cm, the growth is weak, the mycelium is put on light, the conidia are yellow, the medium is colored yellow. . On the Chapek-Doks medium with sucrose, on the 5th day of growth, the diameter of the colonies is 60 3.0-3.2 cm, the growth pattern is similar to on the Chapek medium with starch. The medium is not pigmented. On Gatchinson's medium, on the 5th day of growth, white loose mycelium forms on the surface of the paper65, the medium and a strip of paper are colored lemon yellow. On potato slices, growth is good, mycelium is fluffy, white, lemon conidia is yellow, a slice of potato is painted yellow. Physiological and biochemical characteristics. Assimilates glucose, galactose, maltose, xylose, mannose, sucrose, glycerin, lactose. Fiber is hydrolyzed to glucose, starch hydrolyzes, gelatin does not thin, milk does not coagulate. Uses ammonium, nitrate and protein forms of nitrogen. The strain intensively produces cellulase during growth on a medium with pulp pulp. The maximum accumulation of cellulases for 144 hours. The activity of the C | -enzyme for 4 days is 17-18 units / ml, for 6. days 22-25 units / ml.