RU2180689C2 - Consortium of strains of bacteria rhodococcus erythropolis and yeast saccharomycopsis fibuligera as protein producer in nutrient medium containing alcoholic mash and grain raw - Google Patents

Abstract

FIELD: biotechnology, microbiology. SUBSTANCE: invention proposes the novel protein producer. The latter is a consortium of bacterial strain Rhodococcus erythropolis VKPM-1379 and yeast Saccharomycopsis fibuligera BKPM Y-2173 taken in the ratio = = (1:10)-(1:5). The consortium is cultured in nutrient medium containing alcoholic mash as production waste and grain raw. This provides to utilize food industry waste and protein yield in the end product to 65% simultaneously. EFFECT: new protein producer indicated above, increased yield of protein. 1 tbl

Description

 The invention relates to the microbiological industry, in particular, to the production of protein, and is a new consortium consisting of strains of bacteria and yeast, capable of synthesizing protein on a medium that includes alcohol stillage and cereals.

 Known strain of yeast Candida famata VKPM U-2207, capable of growing on a mixed hydrocarbon-hydrocarbon substrate in a wide range of cultivation process parameters, providing a crude protein content in biomass of up to 45% (RU, patent 2090610). However, this strain, grown as a monoculture, requires the presence in the nutrient medium of a fermentolizate of grain or grain with the addition of n-paraffins, which greatly complicates the process and increases the cost of the finished product due to the need for enzymatic hydrolysis of grain and purification of n-paraffins introduced into the culture medium . In addition, this strain, belonging to the genus Candida, is conditionally pathogenic.

Known yeast strain Saccharomyces cerevisiae VKPM U-760 - a producer of protein and biologically active substances (SU, AS 1575570). This strain is grown on a nutrient medium containing carbohydrates (sucrose, molasses, etc.). The productivity of the process is 2.07-2.57 g of ACB / m ³ • hour. The disadvantage of this strain is the insufficiently high productivity of the process with a sufficiently high flow rate, as well as the need to use carbohydrates in nutrient media.

 Currently, the urgent problem is to obtain protein biomass from sources representing waste from a particular industry for the purpose of their disposal and at the same time cheapen the process of obtaining protein.

 The technical problem solved by the present invention is to develop a consortium of microorganisms for processing a mixture of alcohol stillage and cereals into protein.

 The technical result obtained as a result of the implementation of the invention consists in increasing the content of crude protein in the protein product obtained on the basis of alcohol stillage and grain raw materials to 59-62%.

 To achieve the technical result, it is proposed to use a consortium of bacterial strains Rhodococcus erythropolis VKPM S-1379 and yeast Saccharomycopsis (Eudomycopsis) fibuligera VKPM U-2173, taken in a ratio of 1: 10-1: 5, a protein producer on a nutrient medium containing alcohol distillery grain and raw materials .

 The invention consists in the joint cultivation of the above strains, taken at a ratio of 1: 10-1: 5, on a nutrient medium containing waste from the production of alcohol (alcohol stillage) and waste from the processing of cereal crops or grain.

 The qualitative composition of the consortium makes it possible to utilize starch contained in grain raw materials by splitting it into dextrins and free sugars with the Sacchoromycopsis fibuligera yeast strain. The growth rate of the bacterial strain Rhodococcus erythropolis and its consumption of starch cleavage products determines the strain ratio selected in the consortium.

Cultural and morphological features
Cultural and morphological characteristics of the bacterial strain Rhodococcus erythropolis VKPM S-1379. The strain is isolated from the soil. Forms creamy-colored colonies of rounded shape on meat-peptone agar with a smooth edge. Cells are cocci; culture can form chains of cocci, aerobes. Deposited in the All-Union Collection of Microorganisms under the number S-1379.

 The strain Saccharomycopsis fibuligera VSB-12 was isolated from agricultural waste. On wort agar forms colonies with a smooth edge, 4-10 mm in size, convex profile. Light brown. Cell morphology: oval cells (4-6) - (6-12) microns, separated mycelium, forms ascospores. Physiological characteristics: oval cells (4-6) - (6-12) microns, septic mycelium, forms ascospores. Physiological characteristics: ferment glucose, starch, aerob. Deposited in VKPM under the number U-2173.

Physiological and biochemical characteristics of the consortium
The type of consortium is a symbiosis based on commensalism. Saccharomycopsis fibuligera yeast breaks down starch to form dextrins and free sugars, which are the source of energy and carbon for the bacteria Rhodococcus erythropolis. The ratio of cultures in the inoculum is selected in the range from 1: 5-1: 10, taking into account the growth rate of bacteria Rhodococcus erythropolis and yeast Saccharomycopsis fibuligera.

 Table 1 shows data showing how the protein content in the finished product changes with a change in the ratio of bacterial and yeast cells at the time of seeding of the bioreactor. The protein content was determined by the Kjeldahl method.

Example 1
Mineral salts were added to a nutrient medium containing water, alcohol distillery grain and grain taken in a ratio of 2: 1 by dry weight (total solids content during sowing was 6%):
ammonium sulfate - 3 g / l
monosubstituted potassium phosphate - 2 g / l
magnesium sulfate - 0.5 g / l
manganese sulfate, pentahydrate - 0.02 g / l
ferrous sulfate, heptahydrate - 0.02 g / l
zinc sulfate, heptahydrate - 0.02 g / l
A consortium of bacteria and yeast VSB-12 was seeded in a ratio of 1:10. Cultivation was carried out at T = 30-32 ^o C, pH 5.5, stirring 800 rpm for 12 hours. The result was a product containing 58% crude protein.

Example 2
The cultivation was carried out in the same way as shown in example 1. Bacteria and yeast were seeded in a ratio of 1: 10. A mixture of alcohol stillage and grains taken in a ratio of 1.2: 1 by dry weight was used as a substrate, while the total solids content at seeding was 13%. The result was a product containing 65% crude protein.

Example 3
Cultivation was carried out as shown in example 1. Bacteria and yeast were seeded in a ratio of 1: 5. A mixture of alcohol stillage and grains taken in a ratio of 1.2: 1 was used as a substrate, while the total solids content was 6%. The result was a product with a crude protein content of 57%.

 Based on the experiments, we can conclude that the proposed consortium is a promising protein producer, actively growing on a nutrient medium containing waste products - alcohol distillery grain and raw materials, which ensures the production of high-grade protein feed.

Claims (1)

 A consortium of bacterial strains Rhodococcus erythropolis VKPM S-1379 and yeast Saccharomycopsis fibuligera VKPM U-2173, taken in a ratio of 1: 10-1: 5, is a protein producer on a nutrient medium containing alcohol distillery grain and raw materials.

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