SU1124026A1 - Method for culturing fungus producing protein-enzyme complex - Google Patents

Abstract

A CULTIVATING METHOD FOR A MUSHROOM PRODUCER OF A PROTEIN-ENZYME COMPLEX, which involves the grinding of cellulose-containing raw materials followed by preparation of a nutrient medium containing cellulose-containing raw materials as a source of carbon, sources of nitrogen and phosphorus. source of microelements and water, and by fusing a fungus-producer under it under deep conditions with aeration and mixing of the medium, characterized in that, to increase the activity of the protein-enzyme complex and reduce the energy intensity of the process, Myceliophto and thermophila F are used as a producer fungus -2148i), the cultivation of the fungus is carried out for 1624 hours at 45-47 ° C, continuous aeration of 0.5-0.7 v / v of the nutrient medium and at a stirring intensity of 100-150 rpm, and in the nutrient medium as a carbon source, enter flax Stream, as (L source of microelements and growth substances - flax lobe drains with content, wt.%: Flax on fire 2,0-10,0 Nitrogen source 0,4-0,8 Source of phosphorus 0,1-0,3 Flax lobe drains 20,0-40,0s ts9 About Yu sp

Description

This invention relates to microbiological conditions, in particular, to methods for cultivating microscopic fungi. There is a method of cultivating fungi for 7 days with a rocking chair (220 rpm) on a nutrient medium containing a beetroot as a carbon source, sources of nitrogen and phosphorus, and malt sprouts. The activity of the culture liquid on filter paper 18 u / ml P. Disadvantages of this method are the use of beet oil as a source of carbon - food industry waste, which is a scarce raw material, as well as the duration of cultivation, which requires a considerable amount of energy. The closest in technical essence and the achieved effect to the invention is the method of cultivating the fungus-producer of the protein-enzyme complex, which involves grinding the cellulose-containing raw material with the subsequent preparation of a nutrient medium containing cellulose-containing raw material as a carbon source, nitrogen and phosphorus sources, a source of trace elements and water, and growing a mushroom producer on it in deep conditions with aeration and mixing of the medium. According to a known method, the nutrient medium of the following composition is used: Straw (milling degree. 25%),% 0.5-2 NaNOj, g / l3 KH2PO4, g / l 1 –CN1, g / l 0.5 MgSO, g / l0, 5 FeSO, g / l0, .01 Corn extract,% 0.5 Cultivation of the fungus Aspergillu terreus 17P is carried out at a temperature of cultivation for A day at a constant operation of the stirrer 300 rpm in aeration at a flow rate of 1.5 about / about nutrient medium. As a result, the cellulose activity of the culture liquid is 0.72 mg RV / ml in Na-KMLl, and 0.12 RV / ml in cotton. The indicated composition of the nutrient medium and the conditions of cultivation are proposed for the production of an active cellulose complex, the maximum activity of which is formed by the fungus Aspergillus terreus 17P on the 4th day of cultivation. Data on the content of crude protein in biomass is not given. The disadvantage of this composition of the nutrient medium is that it uses chopped straw as a carbon source, which is a non-localized waste of agriculture. Collecting and transporting straw to ensure an uninterrupted year-round production process imposes on the prime cost a considerable transport cost, which accordingly entails a significant increase in the prime cost of the final product. In the proposed nutrient medium, corn extract is used in a sufficiently large amount (0.5%). The disadvantages of the method of cultivation of the fungus Aspergillus terreus 17P are also the duration of cultivation (4 days), a significant consumption of sterile air (1.5 v / v nutrient medium), which requires a large power consumption. In addition, in order to improve aeration, intensive mixing of the culture fluid (300 rpm) is additionally proposed, which is associated with a high energy consumption, under the specified cultivation conditions requiring a significant energy consumption, and a cellulose complex with low activity is formed. The purpose of the invention is to increase the activity of the protein-enzyme complex and reduce the energy intensity of the process. The goal is achieved there that according to the method of cultivation of the fungus-producer of the protein-enzyme complex, I intend to grind the cellulose-containing pulp with subsequent preparation of a nutrient medium containing cellulose-containing raw material as a source of carbon () yes, sources of nitrogen and phosphorus, a source of microelements and water, and cultivation on it the producer fungus in deep conditions with aeration and mixing of the medium, you use the strain Myceliopht.ora thermopbila F-2148D as a production fungus, growing 1 fnb suschestvl out for 16-24 hours at 45-47 ° C, continuous aeration 0.50 7 v / v of the nutrient medium and stirring at 100 150 intensity. rpm, and in a nutrient medium as a carbon source, an injected flax campfire, as a source of microelements and growth substances, flax lobe streaks at a content, wt.% Flax on the fire 2.0-10.0 Nitrogen source 0.4 -0.8 Source of phosphorus 0.1-0.3 Flax lobe drains 20.0-40.0 The essence of the method is as follows. Flax on the fire (waste from the trapping industry) is used as a carbon source and as a substrate, which is directly biotransformed by a known strain of the microscopic fungus Myceliophtora thermophila F-2148D. As a nitrogen source, is-. ammonium salts (sulfates, nitrates), ammonium fertilizers are used. The source of phosphorus is potassium phosphate salts, phosphate fertilizers. As a source of microelements and organic substances that contribute to biosynthesis, flax lobe drains are used. The composition of flax lobe drains is presented in the table.

Example 1. Myceliophtora thermophila F-2148D was cultured on a prototype nutrient medium), having the following composition: Straw,% 1.0

NaNOj, g / l3

KN2RO, g / l .. 1 KC1, g / l 0.5

M8804, g / l 0.5

FeSO, g / l

0.01

Maize Hbtfi extract,% 0.5 Cultivation lasted 24 hours at 45 ° C, continuous aeration 1.5 v / v of the nutrient medium and at constant

the work of the stirrer 300 rpm As a result, cellulose activity on Na-CMC was 0.8 U / ml, and on filter b5mag, 0.08 units / ml.

The content of crude protein in biomass is 14.5%.

Example 2. M. thermophila F-2148D cultivated on a nutrient medium of the following composition, wt.%: Flax on the fire2.0

(NH4) 2S04C, 4

0.1

Drains of flax lobe 20.0 Water piped water The rest is pH 5.0. The cultivation was carried out for 16 h with continuous aeration (0.6 v / v of the nutrient medium) and with stirring (100 v / m. The activity of the culture liquid was 0.5 units / ml for filter paper, for Na-KMH). 1.7 units / ml The amount of crude protein in the biomes is 25%. Example 3. The nutrient medium for refining M.thermophila F-2148D had the following composition, wt.%: Flax on the fire Superphosphate Flax loops 30.0 Water tap The remaining pH5.3 Mushroom was cultured for 20 hours at 47 with continuous aeration of 0.7% v / v of the nutrient medium and stirring at 120 rpm. ltural liquid on filter paper was 0.4 units / ml, on Na-Kl-lH 1, 6 units / ml, the content of raw prot is 18% in biomass. Example 4. M. thermophila F-2148D was cultured for nutritionally medium of the following composition, wt.% g of flax on a fire 10.0 Ammonium sulfate 0.8 Superphosphate0.3 Flax lobe drains 40.0 Water tap water The remaining pH is 5.5 for 24 hours with continuous aeration of 0.6 v / v nutrient media and mixing intensity 130 rpm. The activity of the culture fluid was on the filter paper 0.2 units / ml, Na-CMC 1, 3 units / ml The amount of protein in biomass is 15%. Thus, the proposed method provides the use of flax fires - the main waste of flax processing industry (60-70% of the production volume), which is currently burned and only a small part of it (10-15%) is used. for the manufacture of Kostroplite (for Furniture), flax lobe drains - currently non-utilizable waste of flax processing industry, which removes the need for wastewater treatment, reduces water consumption for technical purposes and eliminates additional costs for raw materials, nonsification of the production process by reducing the cultivation process 4-6 times and the high cultivation temperature (45 s), which contributes to the accelerated growth of the microorganism and in combination with the low pH of the nutrient medium (5.0-5.5) prevents the possibility of infection by foreign microflora, during the cultivation period of 16-24 hours, cellulose accumulates, which is 2 times greater than the activity of the prototype. During the biosynthesis of the fungus, up to 25% of the crude protein is accumulated in the biomass, which allows it to be used as a feed additive for farm animals. In addition, the proposed method allows to reduce energy consumption by reducing the degree of aeration by 2 times, by reducing the intensity by 2-3 times, and by reducing the duration of cultivation by 4-6 times.

Claims (1)

METHOD FOR CULTIVATING THE MUSHROOM PRODUCER OF THE PROTEIN-ENZYME COMPLEX, which involves grinding cellulose-containing raw materials with the subsequent preparation of a nutrient medium containing cellulose-containing raw materials as a carbon source, nitrogen and phosphorus sources, a trace element and water, and cultivating mushroom under the conditions and mixing the medium, characterized in that, in order to increase the activity of the protein-enzyme complex and reduce the energy intensity of the process, as a fungus strain of myceliophtora thermophila F-2148D is used, cultivation of the fungus is carried out for 1624 hours at 45-47 ° C, continuous aeration of 0.5-0.7 v / v of the nutrient medium and with an intensity of mixing of 100-150 r / min, and Nutrient medium is introduced as a carbon source flax fire, as a source of microelements and growth substances - drains of the lobe holding, wt.%: Flaxseed campfire Source of nitrogen Source of phosphorus Stocks of flax fiber 0.4-0.8 0.1-0.3 20,0-40,0