SU574471A1 - Trichoderma species mouldy mushroom strain-t-producer of cellolytic enzymes - Google Patents

Description

one

This invention relates to microbiology, in particular, new mold fungus strains used to hydrolyze cellulosic plant substrates for silage forage in the production of alcohol, as well as to obtain sugary solutions from the food and wood processing industries.

There are various strains of mold fungi used for the hydrolysis of cellulose-containing plant substrates, for example, the Trichoderma veride strain of the Gene-K-910 producer of cell-lytic enzymes I.

Closest to the invention, the well-known strain of mold fungus Trichoderma lignorum 6c-24, used for the biosynthesis of cellulolytic enzymes 2.

Known strains have an unfavorable activity of cellulolytic enzymes and abundant conidia formation, which leads to environmental pollution and deteriorates working conditions.

The purpose of the invention is to obtain a strain with increased activity of cellulolytic enzymes and weak conidia formation, which, therefore, provides an increase in the yield of the end products of hydrolysis.

The strain of mold fungus Trichoderma specics T-producer of cellulolytic enzymes isolated from the soil and is stored in the crop collection of the All-Union Research Institute of Genetics and Breeding of Industrial Microorganisms under the number CMPA1-83.

The activity of cellulolytic enzymes of the strain is stabilized by long-term cultivation on an agar medium with taurine (2-aminoethanesulfonic acid) with a concentration of 0.001%.

Strain Trichoderma species T-producing cellulolytic enzymes are cultured under superficial conditions on sterile, solid, moisturized to 64-65% nutrient medium.

The cellulose activity of the ferment in the final culture is 400-500 u / g of dry culture.

The cellulose activity d is determined by hydrolysis of the filter paper according to a known method.

In the finished culture for 48-60 hours, the growth of conidia is almost absent.

Morphological signs. Mycelium is colorless, prostrate. Growing turf or cobweb. Sods are cushion-shaped, convex, first white, then yellow green and dark green. Conidiophores in the form of lateral branches of the hyphae of the aerial mycelium, for the most part forked or ternate, colorless. Sterigms are bottle-shaped or conical, slightly curved, arranged in 2-3 or single or 6.5-13.5x3.5-4 microns in size. Conidia are ovate-spherical in diameter 3.5-5.0 (5.5) μm, collected in the head, in the mass of white, yellow-green or dark green.

Cultural features. Synthetic environment of čapek with sucrose. Crops prick. After 7 days of growth at a temperature of 28-30 ° C, the colonies are prostrate, the growth is heterogeneous, sometimes by turf. The diameter of the colonies is 9-10 cm. Initially, the colorless stele of the gtium mycelia develops. Aerial mycelium thin, slightly elevated, colorless. Conidia in the form of green and white pads, located in the center. The medium turns bright yellow and forms a yellow pigment that diffuses into the medium.

Wort agar (4.8 balling + 1% shredded filter paper). Colonies prostrate, after 4-5 days of growth at a temperature of 28-30 ° C to 70 mm. Aerial mycelium is pressed, and sometimes spider-like, growth is heterogeneous, sometimes by turf. On the 5-6th day of growth, conidia appear along the edges, the income of the schne to the center of the colony, initially light, then with a greenish tint. The reverse side of the agar is colored bright yellow. Forms a yellow pigment that diffuses into the medium.

Medium composition,%: ground filter paper-1; (NH4) 2SO4 - 0.15; NaNOs -0.15; KH2P04 -0.15; MgSO4-7H20-0.05; 10% wheat bran extract 300 ml; tap water up to 1 l plus 2% agar. Sow a prick. Colonies prostrate, mycelium sometimes cobwebby, sometimes turf. Conidia appear on the 4th – 5th day along the edges, then grow but the whole colony. The reverse side is painted in a slightly yellow color.

Synthetic environment of čapek with starch (medium Gause-1). Sow a prick. After 10–12 days of growth at 28–30 ° C, the diameter of the colonies is 4–5 mm, and at the site of the injection for 12–15 days conidia form in the form of rare white or pale green with a yellowish tinge of nodushech. The medium is an intense yellow.

Sliced potatoes. Sowing stroke. After 14–15 days of growth at 28–30 ° C, the colon covers the entire surface of the slice, a spike, tussock turf, painted in a light green color with separate patches of dark green or light yellow color, copious formation is abundant, the substrate on the reverse side is intensely colored bright yellow color.

Physiological signs. Relation to carbon sources. Assimilates glucose, galactose, lactose, maltose, xylose, mannose, sucrose, glycerin.

Cellulose hydrolyzes to glucose, starch hydrolyzes very weakly.

Relation to the source of nitrogen. Pitrates recovers by 30 days of incubation.

Gelatin is not diluted after 30 hours of incubation.

Peptonized milk is very weak at pH

Claims (2)

1. USSR author's certificate No. 39522, cl. C 12D 13/10, 1971. 2. USSR author's certificate number 482494, cl. From I2D 13/10, 1975.