SU1418337A1 - Method of cultivating yeast - Google Patents

Abstract

This invention relates to the microbiological industry and relates to a method for cultivating yeast. The aim of the invention is to increase the content of phenylalanine, histidine and threonine in biomass. The yeast is cultivated on n-alkane-containing hydrocarbons and an aqueous nutrient medium under aerobic conditions and mixing of the medium — by regulating the specific input of aeration energy and mixing in the range of 3.5–6.0 kW / t of medium. In the process of cultivation, the ratio of the concentration of n-alkanes to energy input is in the range of 3–10 and the rate of transition of n-alkanes from a mixture of hydrocarbons to cells in the range of 2.5–5.0 kg / ton of nutrient medium. The yeast strain Candida salmonicola BWA-779 is used as a yeast producer. 1 hp f-ly. (L

Description

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with so

This invention relates to the microbiological industry and relates to a method for cultivating yeast with an increased proportion of essential amino acids, primarily phenylalanine, histidine and threonine.

There is a method of cultivating microorganisms with adjustable process parameters. Yeast Totulopsis and Candida are cultivated on the oil fraction in the apparatus with a stirrer and aeration with a stirrer speed of 2000 and an air supply of 400 l / h. In this case, a protein product is obtained (French Patent No. 1297810, Cl. F 01 D, 1980).

 The disadvantage of this method is that the input of energy is very high due to mixing and aeration and biomass is obtained, with an indefinite composition of amino acids.

The closest to the proposed method is the cultivation of yeast on a nutrient medium containing n-alkanes as a carbon source, sources of nitrogen, phosphorus, mineral salt and trace elements under conditions of aeration and mixing, followed by release of biomass (USSR author's certificate No. 413183, CL 12 N 1/09 (1971).

In order to improve the cultivation conditions, the energy consumption of the stirrer is varied in steps. Initially, the stirring rate increases as long as the CO concentration in the exhaust air is reached in the amount of 3-11%. After that, the input of energy is reduced, and the air supply to the device increases.

The disadvantage of the known method is that it is impossible to stably carry out the process of obtaining biomass with a definite content of essential amino acids, including an increased content of phenylalanine, histidine and threonine.

The purpose of the invention is to increase the content of phenylalanine, histidine and threonine in the biomass.

The proposed method consists in that yeasts are cultivated on hydrocarbons in the presence of the necessary mineral elements in aerobic fishing, while aeration and mixing are carried out with a specific energy input of 3.5-6.0 kW / ton of medium, and

The cultivation process is carried out at a ratio of the concentration of n-alkanes in a nutrient medium to the specific input of energy in the range of 2.5-5.0 kg / t.

At the same time, a positive effect when using the Candida salmonic yeast BSB-779 (stored in the collection of the central museum of microorganisms of the Institute of Scientific Research Institute-Genetics of the General Directorate of the Microbiological Industry under the USSR Council of Ministers under the number of CMPM U-196) than with other yeast strains.

The yeast species of Candida, Trichos patches or others are cultivated in a continuous process in a fermenter equipped with mixing and aeration devices. Petroleum distillates containing 10-25% n-paraffins are used as carbon source. Ammonia nitrogen serves as a nitrogen source of nitrogen, and various salts containing these elements can be a source for phosphorus, potassium, magnesium and trace elements.

The concentration of the applied oil distillate in the fermentation medium is set so that the concentration of n-paraffins (G) is 19-39 gAqg.

 Concentrations of individual nutrients and trace elements are in the fermenter, mg / kg: N 2700i1500; P 370 + 180; K 625t375; Mg 40t25; Fe 3.95t2.05; Zn 9,015,5; Mn 5.75t3.25; Cu 2,3t1,75.

The yeast is cultivated at a dilution rate of 0.2-0.6 h, pH 2.5-4.5, temperature 32-40 ° C and aeration rate 80-160.

The total specific energy input for mixing using a mixing device and for aeration using a fan, a suction mixer, or other device that simultaneously mixes and aerates is 3.5-6.0 kW / tonne of medium. In this case, the total specific energy input is determined by the following formula

N

Milln.

N i + N2j: Nj MF

N specific total specific consumption

energy, kW / t; N, - power consumption (net)

for mixing, kW / t; N - power consumption (net)

for aera 1i, kW;

31418337

Nj is the energy consumption (net). The total energy input (net) is a part of the possible additional 33 kW, and the total specific input

energy 33 / 5.5 6.0 kW / t.

As a carbon source, an oil distillate with a paraffin content of C of 0.145 kg / kg was used. The distillate concentration was

mixing plants, kW; Мр - filling volume of the fermenter, t.

With the establishment of the total specific energy input at the same time,

at the inlet of the fermenter 0.2 kg / kg

A petroleum distillate with a paraffin content of C of 0.145 kg / kg was used as the carbon source. The distillate concentration was

at the inlet of the fermenter 0.2 kg / kg

carbon concentration which corresponds to the paraka concentration so that the ratio S

Pai / N

VAf / itH of the inlet 29 kg / ton.

VAf / itH was in the range of 3.0-10, O, and the concentration of n-paraffins used relative to the total inflow into the fermenter, kg / t.

The rate of transfer of paraffins from the hydrocarbon mixture into the cells is set at 2.5-5.0 kg / t h. The transition rate is determined by the formula

15

The concentration of nutrients was mg / kg: N 2600; P 340; K 480; Mg 24; Fe 3.0; Zn 7.1; C 1,2.

The cultivation was carried out at. pH 3.5-3.7, temperature 309-311 K, dilution rates 0.25 h and skog

Par Chg -T | -R "T) -0-103 (/ ,.)

kv

where C

. R..

p ", 0.2 (1-1GO.). about 25-103,

 Pair

 4.4 kg / t.h.

thirty

growth aeration 120 m.

During cultivation, the concentration of paraffins in the oil distillate and the rate of paraffin transition were determined. It is found C P 0.063 kg / kg, these are obtained,

- hydrocarbon concentration -.

mixture in the inflow of fermenter 25 kg, kg / kg; and C is initial and relevant

the concentration of paraffins in the hydrocarbon mixture, kg / kg;

D is the flow rate, h. When regulating the cultivation process within the prescribed limits for

specific energy input for , „„ „„

neither the concentration of paraffin to specific. ,, was 5.4% of histidine 3.9%

energy input and speed paraffin transition process is stabilized and biomass is obtained with an increase in the ",". ,

Example 2. Candida Yeast

salmonicola BWA-779 (CMPM U-196) were likewise likewise as in Example 1; at. fermentor filling volume 75 tons. The flow rate was 0.25, the intensity of aeration 87 m

in ns. Total energy input (sum, per iz. mixing and aeration) is, dals at 375 kW, respectively

Separated and purified according to known methods, the biomass contained 62.4% of the blue protein or 52, Zy. pure protein. Phenyl content

 35 and threonine 5.9% relative to the sum of amino acids.

content of essential amino acids.

The yeast suspension, which is continuously withdrawn from the fermenter, is further processed according to known methods. The final product (biomass) contains,%: cheese protein 59-62,5; pure protein 45.0-52.3; phenylapanine to 5.7; histidine to 4.3 and. Treynin up to 6 relative to the sum of amino acids.

 VAcKiH kW / t. As coal -:

; oil source was used

distillate with n-paraffin content.

Example 1. Candida salmonicola yeast (CMPM U-196) of BWA-779 was cultivated in a fermenter with a filling volume of 5.5 tons and with intensive aeration of 120 m in ns / t h. The fermenter is equipped with a stirrer, a pneumatic mixer and an external circulator cooling system.

And the one that corresponds to the concentration of the pairAf / itHem aa a

nao15

finov inlet 29 kg / t.

The concentration of nutrients was mg / kg: N 2600; P 340; K 480; Mg 24; Fe 3.0; Zn 7.1; C 1,2.

The cultivation was carried out at. pH 3.5-3.7, temperature 309-311 K, dilution rates 0.25 h and skog

p ", 0.2 (1-1GO.). about 25-103,

 Pair

 4.4 kg / t.h.

thirty

. , „„ „„

Separated and purified according to known methods, the biomass contained 62.4% of the blue protein or 52, Zy. pure protein. Phenyl content

. ,, was 5.4% histidine 3.9%

  „„,. ,

 35 and threonine 5.9% relative to the sum of amino acids.

 VAcKiH kW / t. As coal -:

oil source was used

distillate with n-paraffin content.

j-. Speech - 0.155 kg / kg at a concentration.

di stil that Cj 0.18 kg / kg. The paraffin concentration in the medium was thus 5 27.9 kg / ton. All other parameters correspond to the example 1.

During cultivation, the actual concentration of n-paraffins in the distillate was 0.074 kg / kg.

 I

The values of N ,,, the ratio of SQ nodozhilis thus when

street 5,0 kW / t; S vAPNkH 5.6; np "v. 0.18 (1) 0.25-103;

3.9 kg / T.

4.2, temperature 305-307 K and aeration rate 87 m in n, s./t h.

The actual concentration of n-alkanes in the distillate was 0.07.3.

Process indicators were following s

 Par

Separated and purified biomass

NVACAIH 5.4 kW / t; 10 5.6 ;.

Podr- 5.0 .V

Separated and purified biomass contained,%: With protein 60.1; true protein 47.2; phenylalanine 4.8; contained,%: crude protein 59.1; is-15 histidine 3.5 relative to the amount of the amine protein 51.9; phenylalanine 5.2; but acids

histidine 4.1; threonine 5.8 relative to Example 5. Drbzhi Lodderomy- but the sum of amino acids.ces .elongisporus 128 (C.guilP p and m ep 3. Yeast G.salmoni-liarmondii Druzba) were cultivated with BCA-779 cola (CMPM U-196) cultivar - 20 fermenters with a filling volume of 20 kg were similar to those of example 1 in a ferrite system as in example 4. The filling volume of 5 kg was used. The rate of flow was not 0.22 h. - - The total energy input was 19 watts. As a carbon source, an oil distillate with a concentration of n-alkanes of C 0.138 was used.

The distillate concentration was in the flow of the fermenter C 0.26 and

appropriately S

Pci

35.9 kg / ton Dru- 30

The flow rate was 0.2 and the aeration rate was 150 liters per NS / kg-h.

The total energy input for mixing and aeration was at 86 watts. A carbon distillate with a C of 0.12 was used as the carbon source.

The distillate concentration was C = 0.158. The current concentration of paraffins was established at C 0.046.

These parameters are identical to Example 1.

The current concentration of n-alkanes in the effluent oil distillate was C "0.084.

Thus the process indicators are as follows:

N VAEAOH 3.8 kW / t;

 Rig

/ N

leln, 9, 5;

pd, y 3.4 kg / t-h

The separated and purified biomass contained,%: crude protein 60.1; true protein. 51.3; phenylalanine 5.7; histidine 4.3; threonine 5.9, relative to the sum of amino acids.

Ir and Mer 4. Lodderonr; - ces elongisporus Il-ffiT-H 128 (Candida guilliermondii Druzba) yeast were cultivated in a fermentor with a filling volume of 75 tons as in Example 2. The total energy input for mixing and aeration was 405 kW. A petroleum distillate with a concentration of n-alkanes C O ,, 19 was used, the concentration of the distillate in the flow medium being C = 0.158. The paraffin concentration in the inflow was thus 30 kg / ton. The cultivation was carried out at a pH of 4.04, 2, a temperature of 305–307 K, and an aeration rate of 87 m in n, s./t h.

The actual concentration of n-alkanes in the distillate was 0.07.3.

Process indicators were following s

liermondii Druzba) cultured with a fermentation tank of 20 kg in volume 0 as in Example 4. 5

0

five

0

five

0

five

The flow rate was 0.2 and the aeration rate was 150 liters per NS / kg-h.

The total energy input for mixing and aeration was at 86 watts. A carbon distillate with a C of 0.12 was used as the carbon source.

The distillate concentration was C = 0.158. The current concentration of paraffins was established at C 0.046.

Process indicators were as follows:

N, de 4.3 of W / kg; S / NvAtAbH 4.4;; - Pr 2.5 kg / t-h. The separated and purified biomass contained,%: crude protein 61.3; true protein 45,4; phenylalanine 4.5; histidine 3.0; threonine 5,1 relative to the amount of amino acids.

Example 6. Lodderomyes dongisporus IMET-H 128 yeast (C. guilliermondii Druzba) was cultivated as in Example 5. The total energy input for stirring and aeration was 120 W, and the flow rate was 0.275 h. Other parameters are identical to Example 5. The current concentration is -alkanes in distillate established at a CP of 0.045.

The process indicators were as follows:

  6 W / kg; h; / Cudgd, „3,2; Prd 3., 4. kg / t-h

. 7 .1Д

The separated and purified biomass contained,%: crude protein 60.1; juvenile protein 46.2; phenylalanine 5.0; histidine 3,2 relative to the amount of amino acids.

Example 7. Yeast Lodderomyces elongisporus IMET-H 128 (C.guillie - mondii- Druzba) was cultivated as in Example 5. General input. energy for mixing and aeration was 60 watts. all other parameters correspond to Example 5. The current concentration of n-alkanes in the distillate was set at 0.049.

Process indicators were as follows:

NVAEAW 3W / kg; sP VNvAeA.H 6.3; Prague 24 g / kg.h,

The separated and purified biomass contained,%: crude protein 59.8; true protein 45,8; phenylalanine 4.1; histidine 2.8; threonine 4.3 relative to the amount of amino acids.

Thus, by setting the fermentation process on the parameters according to the proposed method, the content of the essential amino acids potassium phenylalanine, histidine and treoni was achieved without additional costs.

by up to 5.7; 4.3 and 5.9% with the use of yeast C, salmpnicola BWA-779 and up to 5.0; 3.5 and 5.10% when cultivating g Lodd yeast. elog. IMET H 128 (S. guilliermondii Druzba).

Claims (2)

Invention Formula The method of cultivation of yeast on a nutrient medium containing n-alkanes as a carbon source, sources of nitrogen, phosphorus, mineral salts and trace elements under conditions aeration and mixing, followed by the distribution of biomass, characterized in that, in order to increase the biomass content of phenylalanine, histidine and threonine, aeration and mixing is carried out at a specific energy input of 3.5-6.0 kW / ton of medium, and the growing process is carried out at a ratio of the concentration of n-appanes in a nutrient medium to the specific energy input, within 3.0-10.0 and the transition rate n-alkanes to cells in the range of 2.5-5.0. 2. A method according to claim 1, characterized in that it is cultivated. the strain of the yeast Candida salmonicola BWA-779.