CN110885766A

CN110885766A - Biological agent for degrading organic matters in amino acid mother liquor

Info

Publication number: CN110885766A
Application number: CN201911281380.XA
Authority: CN
Inventors: 罗玮; 王小平; 翟德亮
Original assignee: Zhao Lankun
Current assignee: HULUNBEIER NORTHEAST FUFENG BIOTECHNOLOGIES Co.,Ltd.
Priority date: 2019-09-16
Filing date: 2019-12-13
Publication date: 2020-03-17
Anticipated expiration: 2039-12-13
Also published as: CN110885766B

Abstract

The invention belongs to the technical field of biology, and discloses a biological agent for degrading organic matters in amino acid mother liquor, which is prepared by the following processes: respectively inoculating beer yeast, rhodotorula glutinis and Angel yeast into a seed culture medium for culture, and then uniformly mixing to obtain a bacterial suspension; mixing the bacterial suspension and diatomite, stirring, drying at 20-22 deg.C under low temperature, drying to water content of 8-10%, and refrigerating. The biological agent has reasonable compatibility and good synergistic performance.

Description

Biological agent for degrading organic matters in amino acid mother liquor

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a biological agent for degrading organic matters in amino acid mother liquor.

Background

China is a large country for producing and consuming amino acids, and a large amount of wastewater containing amino acids is often discharged in the production of chemical industry, pharmacy, food and other industries, and taking glutamic acid production as an example, China produces 230 ten thousand tons of glutamic acid every year and discharges more than 1900 ten thousand tons of high-concentration acidic amino acid waste mother liquor. COD and SO in waste mother liquor₄ ^2-、NH₄ ⁺the-N and thallus content is high, the thallus is acidic, the self-cleaning capacity of the water body can be obviously reduced after the thallus is discharged into the water body, the growth of microorganisms in the water body is hindered, the environment is seriously polluted, and the treatment difficulty is higher. In order to solve the problem of wastewater pollution and effectively utilize the wastewater resources, in the traditional production process, mycoprotein in the amino acid waste mother liquor is extracted to be made into feed for sale, and the waste mother liquor after protein extraction is concentrated and utilized to prepare fertilizer for feedback agriculture by a slurry spraying granulation technology.

The spray granulation process of the amino acid fermentation waste liquid is an effective way for effectively treating high-concentration waste water by amino acid production enterprises, realizing circular economy and increasing the economic benefits of the enterprises, but in the spray granulation process of the fermentation waste liquid, polluted smoke containing a large amount of water and organic matters is generated, and secondary pollution to the environment is easily caused. The smoke has serious peculiar smell, influences the normal life of the surrounding masses, becomes a problem to be urgently solved by the local government, and is a great technical problem which puzzles the development of the fermentation industry for a long time.

Therefore, the method can reduce the problem of secondary pollution of the flue gas, improve the product performance or quality of the feed and even the fertilizer, and has important significance for the green and healthy development of enterprises. Fufeng takes precedence in the industry to remove more than 80% of smoke dust and peculiar smell by adopting a spray adsorption and plasma technology, so as to achieve the emission reaching the standard, but the equipment adopts fermentation wastewater to carry out spray cooling in a Venturi spraying process, the spray amount is large, the water temperature is high, and a large amount of water vapor is formed secondarily; although the electrostatic dust removal equipment is adopted to effectively remove the solid particles in the waste flue gas, the solid particles in the waste flue gas at the outlet are higher; the water vapor is used for wrapping solid particles which are not completely removed and trace organic matters with the flavor of scorched meat in the concentrated solution, and the organic matters are discharged together through the chimney, thereby causing influence on the environment.

Based on the above situation, the present invention provides a research subject for analyzing and researching the prior waste liquid from amino acid production, and finding out that organic matters such as reducing sugar and amino acid in the waste mother liquid from amino acid fermentation are one of the main factors causing the odor of the spraying granulation process.

The waste mother liquor of glutamic acid production, i.e. the waste water of monosodium glutamate contains various amino acids, residual sugar, fermentation intermediate products, ammonium salt and the like, and is expressed as high COD and high NH₃-N. According to research, the waste mother liquor contains various nutrient components, so that a large number of microbial flora are bred in the storage process of the waste mother liquor in the storage pool, and along with the accumulation of the running time of the storage pool, the dominant microbial flora suitable for the low-pH and high-salinity severe environment of the waste mother liquor is formed in the storage pool. The research center of Fufeng group has conducted a special study on the subject (Lehaiyan, Xuzhouhua, etc. 'research on separation, screening and growth characteristics of dominant bacteria in monosodium glutamate wastewater', fermentation science and technology communication, 2008, No. 4), and the results show that: in the monosodium glutamate wastewater, a large number of microbes of different species grow, and under certain ecological conditions, the growth of various microbes is in a balanced state; the dominant growth bacteria keep dominant growth in the wastewater, the proportion of the growth amount to the total number of microorganisms in the water body is the largest, the external conditions of the dominant bacteria are changed through research, the growth activity of the dominant bacteria is improved, and the degradation effect on wastewater pollutants is greatly improved.

The national emphasis laboratory of environmental water chemistry of the ecological environmental research center of the Chinese academy of sciences uses batch experiments to perform a dehydrogenase activity (DHA) test on a group of yeast mixed floras screened from high-concentration monosodium glutamate wastewater (black and bright, et al, continuous bench scale for treating high-concentration monosodium glutamate wastewater by using yeast, environmental science, 2002, 2 nd stage). The results show that when the bacterium is used for treating the monosodium glutamate ion-exchange tail liquid with COD, sulfate radicals and ammonia nitrogen close to 20000 mg/L, the DHA value of the bacterium is higher than that of the monosodium glutamate ion-exchange tail liquid diluent in the first 36h, and the influence of high-concentration ammonia nitrogen and sulfate on the activity of the saccharomycetes is not obvious.

Yeast, a unicellular microorganism, is mostly present in a sugar-rich environment, requires similar nutrients as other living organisms, has a set of intracellular and extracellular enzyme systems like bacteria, and is a heterotrophic cell for breaking down large molecular substances into small molecular substances that are readily available for cellular metabolism. Yeast is harmless and easy to grow, can grow within the pH range of 3.0-7.5, can live in oxygen or oxygen-free environment, decomposes sugar into carbon dioxide and water under the aerobic condition, and enables yeast to grow fast. In the absence of oxygen, yeast break down sugars into alcohol and carbon dioxide. The yeast has low requirement on temperature, the optimal growth temperature is generally 20-30 ℃, and the culture, production and application are convenient. Yeasts grow in both aerobic and anaerobic environments, i.e., facultative anaerobes. In the presence of oxygen, the yeast decomposes the sugar into carbon dioxide and water and grows faster; in the absence of oxygen, yeast break down sugars into alcohol and carbon dioxide. However, screening suitable yeast species to improve degradation efficiency is a difficult point of research.

Disclosure of Invention

In the prior patent technology of the applicant, a process for degrading organic matters in amino acid fermentation waste liquid by using a composite microbial inoculum is used for degrading the organic matters by using composite yeast, namely, bacterial suspension is inoculated, however, in the actual operation, the bacterial suspension is often required to be dried to prepare a biological preparation for storage and subsequent use, and on the basis, the invention provides the biological preparation for degrading the organic matters in amino acid mother liquor.

The invention is realized by the following technical scheme:

the biological agent for degrading the organic matters in the amino acid mother liquor is prepared by the following process:

respectively inoculating cerevisiae Fermentum, Rhodotorula glutinis and Angel yeast to seed culture medium, and culturing to concentration of 10⁸cfu/ml, and then uniformly mixing to obtain a bacterial suspension; mixing the bacterial suspension and diatomite, stirring, drying at 20-22 deg.C under low temperature, drying to water content of 8-10%, and refrigerating.

Further, the biological agent is prepared according to the following process:

respectively inoculating cerevisiae Fermentum, Rhodotorula glutinis and Angel yeast to seed culture medium, and culturing to concentration of 10⁸cfu/ml, and then uniformly mixing according to the equal volume proportion to obtain a bacterial suspension; mixing the bacterial suspension and diatomite at a mass ratio of 1:1, stirring at 100rpm for 3min, drying at low temperature of 20-22 deg.C to water content of 8-10%, and refrigerating at 4 deg.C.

Preferably, the preparation method of the seed culture medium comprises the following steps: taking 10g of yeast extract powder, 20g of peptone, 20g of glucose and 20g of agar, adding water, diluting to 1000ml, sterilizing at 121 ℃ for 20min, and naturally cooling to obtain the yeast extract.

More preferably, the lager brewing yeast is ATCC 9763.

More preferably, the Rhodotorula glutinis is ATCC 32765.

The beneficial effects achieved by the invention mainly comprise but are not limited to the following aspects:

when the amino acid waste mother liquor is subjected to spray granulation to prepare the organic-inorganic compound fertilizer, the temperature is high, and reducing sugar, amino acid, organic matters and the like are coked to generate burnt odor, so that the harmful effect on the surrounding environment is generated, and the nutritional ingredients are damaged. The nutrient components can be used for microbial fermentation, and the content of reducing sugar, amino acid and organic matters in the waste mother liquor is reduced to reduce the burnt smell and concentration thereof generated in the process of spraying granulation.

The state of the microorganism growing in the spent liquor determines the effect of the microorganism on the utilization of the spent liquor. Therefore, the selection of the bacterial strain which can grow in the harsh environment of the amino acid waste mother liquor with low pH and high COD for fermentation culture has a decisive significance for removing the burnt odor components.

According to the invention, a large number of experiments are carried out to screen out the composite biological agent suitable for growth under severe conditions of low pH, low nutrition, high salt and the like, and the biological agent is utilized to degrade organic matters such as reducing sugar, amino acid and the like in the amino acid waste mother liquor, so that the sugar content and the whole organic matters are greatly reduced, and the substances causing burnt odor in the process of spraying granulation are reduced.

Detailed Description

Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the products and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations and modifications, or appropriate alterations and combinations, of the products and methods described herein may be made and utilized without departing from the spirit, scope, and spirit of the invention. For a further understanding of the present invention, reference will now be made in detail to the following examples.

Example 1

Analysis of amino acid fermentation wastewater

As can be seen from the test report (Table 1) of the amino acid waste mother liquor of various batches, the waste mother liquor has a high acidity (pH 3.73) and contains various amino acids, residual sugars, fermentation intermediates, ammonium salts and other components.

TABLE 1 detection results of amino acid waste mother liquor

And (4) selecting the most suitable yeast single strain.

In order to examine the degradation performance of various yeast strains on reducing sugar in the amino acid waste mother liquor, four strains of A1 (beer yeast ATCC9763), A2 (Rhodotorula glutinis ATCC 32765), A3 (Candida utilis ATCC 22023) and AL (Angel yeast) obtained by culturing (the culture medium is shown in Table 2) are respectively inoculated into the amino acid waste mother liquor according to the inoculation amount of 10 percent, the rotating speed of a shaking table is controlled, the culture is carried out for 48 hours, and sampling is carried out once every 12 hours to detect the change of the concentration of the reducing sugar in the waste mother liquor. The pH value is measured by a Metler pH meter, the reducing sugar content is titrated by a Fehling reagent, and the glucose content is measured by a biosensor.

TABLE 2 basal case of slant Medium

(Note: adding glucose when the slant culture medium is cooled to about 50 ℃ and shaking to form a slant)

(1) Application of strain A1 to reducing sugar in waste amino acid mother liquor

As shown in Table 3, Yeast Strain A₁After 48 hours of culture, the OD value of the wastewater is increased from 0.443 to 0.535 on average, and the OD value is increased by 0.092; the pH value of the wastewater is increased from 4.025 to 4.328 on average and is increased by 0.303; the reducing sugar is reduced from 0.475 percent to 0.220 percent on average and is reduced by 53.68 percent.

TABLE 3 detection of the sugar-reducing shaking flask test procedure of the strain A1 wastewater

(2) Strain A₂Application of reducing sugar in waste amino acid mother liquor

Table 4 shows, Yeast Strain A₂After 48 hours of culture, the OD value of the wastewater is increased from 0.440 to 0.534 on average, and the OD value is increased by 0.094; the pH value of the wastewater is increased from 4.018 to 4.315 on average and is increased by 0.294; the reducing sugar is reduced from 0.463% to 0.198% on average and reduced by 57.24%.

TABLE 4 Strain A₂Detection of wastewater blood sugar reduction shaking experiment process

(3) Strain A₃Application of reducing sugar in waste amino acid mother liquor

Table 5 shows, Yeast Strain A₃After 48h of culture, the OD value of the waste mother liquor is increased from 0.444 to 0.536 on average, and the OD value is increased by 0.092; the pH value of the wastewater is increased from average 4.033 to 4.325, and is increased by 0.292; the reducing sugar is reduced from 0.443% to 0.230% on average and is reduced by 48.08%.

TABLE 5 Strain A₃Detection of wastewater blood sugar reduction shaking experiment process

(4) Strain A_LApplication of reducing sugar in waste amino acid mother liquor

Table 6 shows, Yeast Strain A_LAfter 48h of culture, the OD value of the wastewater is increased from 0.435 to 0.536 on average, and the OD value is increased by 0.104; the pH value of the wastewater rises from 4.043 to 4.308 on average and rises by 0.265; the average reducing sugar content is reduced from 0.478% to 0.195%, and the reducing sugar content is reduced by 59.21%.

TABLE 6 Strain A_LDetection of wastewater blood sugar reduction shaking experiment process

(5) Comparison of reducing sugar reducing effects of four yeasts

Comparing the blood sugar reducing performance of four yeasts, the result shows that A_L>A₂>A₁>A₃Thus, it can be seen that yeast strain A_LThe reducing sugar reducing effect of (2) is the best.

Example 2

The effect of reducing sugar is reduced by the compatibility of the yeast.

Selecting A with better hypoglycemic effect_L、A₂、A₁The three strains are compatible to improve the blood sugar reducing efficiency.

Combination 1: a. the_L＋A_2；

And (3) combination 2: a. the₂＋A_1；

And (3) combination: a. the_L＋A_1；

And (4) combination: a. the_L＋A₂＋A_1。

And (4) optimizing growth conditions.

In order to determine the optimal growth pH value and the optimal conditions for reducing the sugar of the yeast strain, preparing a seed culture medium according to the table 7, heating and dissolving, subpackaging test tubes, sterilizing at 121 ℃ for 20min, and taking out the test tubes when the test tubes are hot to prepare the seed culture medium; inoculating each strain to seed culture medium, and culturing to concentration of 10⁸cfu/ml, each strain was as per 1:1 to obtain a bacterial suspension. Then preparing fermentation culture medium, adding amino acid fermentation waste liquor into the container, addingAdding 0.1% magnesium sulfate, separately removing, adding 0.5% ammonium sulfate, 0.3% potassium dihydrogen phosphate, and 0.02% calcium chloride, adjusting pH to 4.0, sterilizing at 121 deg.C for 20min, inoculating bacterial suspension according to 10% inoculum size, and culturing at 34 deg.C for 24-48 h. The two different batches of waste liquid are treated respectively to verify the effect.

TABLE 7 basic conditions of the seed culture Medium

Medicine and food additive	Quality of	Manufacturer of the product
			Yeast extract powder	10 g	Ao Bo xing
Peptone	20 g	Japanese
			Glucose	20 g	Is administered orally
Agar-agar	20 g	An inlet
			Water (W)	1000 mL	Fufeng (Chinese character of' Fufeng

TABLE 8 basic conditions of the fermentation media

Medicine and food additive	In proportion%	Manufacturer of the product
			Magnesium sulfate	0.1	Individually distribute
Ammonium sulfate	0.5	Adjusting pH to 6.5
			Potassium dihydrogen phosphate	0.3	Adjusting pH to 6.5
Calcium chloride	0.2	Adjusting pH to 6.5

The waste liquid is treated by the yeast combination, and specific experimental data are shown in table 9.

TABLE 9 Experimental Process testing of different combinations of yeasts

Therefore, compared with a mode of combining two yeasts, the combination 4 adopts the compatibility of three yeasts, has the best degradation effect on organic matters and reducing sugar in the waste mother liquor, is obviously superior to the combination of the compatibility of two strains, shortens the treatment time, and can achieve the better treatment effect within 36 hours; therefore, the yeast is utilized to reduce the content of reducing sugar, so that the whole organic matter content of the waste mother liquor can be effectively reduced, and the burnt odor concentration generated in the process of spraying granulation can be effectively reduced.

Example 3

preparing a seed culture medium: taking 10g of yeast extract powder, 20g of peptone, 20g of glucose and 20g of agar, adding water, diluting to 1000ml, sterilizing at 121 ℃ for 20min, and naturally cooling to obtain the yeast extract powder;

The using method comprises the following steps: taking out the biological agent, activating, adding into the mother liquor according to a certain proportion, and treating for more than 24 h. After the mother liquor is treated, yeast can be collected and used for preparing yeast extract or yeast powder.

Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. The biological agent for degrading the organic matters in the amino acid mother liquor is prepared by the following process:

2. The biological agent according to claim 1, wherein the biological agent is prepared by the following process:

3. The biological agent according to claims 1-2, wherein the method of preparing the seed culture medium is: taking 10g of yeast extract powder, 20g of peptone, 20g of glucose and 20g of agar, adding water, diluting to 1000ml, sterilizing at 121 ℃ for 20min, and naturally cooling to obtain the yeast extract.

4. A biological preparation according to claims 1-3, characterized in that said brewer's yeast is preferably ATCC 9763.

5. A biological preparation according to claims 1-3, characterized in that the rhodotorula glutinis is preferably ATCC 32765.