Harmless treatment method for steroid hormone mushroom dregs

Abstract

The invention discloses a method for harmlessly treating steroid hormone mushroom dregs, and belongs to the technical field of biomass resource utilization. The preparation method comprises the following steps: mixing an androstenedione-containing mushroom dreg raw material serving as a substrate with anaerobic activated sludge; the mass ratio of the androstenedione-containing mushroom dreg raw material to the anaerobic activated sludge is (1: 3)-(1: 1); and carrying out sealed anaerobic fermentation for 28-32 days in an inert atmosphere at the fermentation temperature of 35-40 DEG C or 50-60 DEG C, and collecting biogas. According to the method, the materials are pretreated at medium temperature and high temperature, the fermentation temperature and time are controlled, and dissolved oxygen in the materials is removed in a nitrogen blowing manner, so that volatile fatty acid in the materials is converted into methane, and accumulation of biogas and elimination of hormone are realized. The androstenedione mushroom dregs with low C / N ratio are used as a single substrate and do not need to be blended with high-carbon-content organic matters for fermentation, so that the cost is saved, the effect is continuous, and the fermentation conditions are easy to control.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomass resource utilization, and particularly relates to a method for harmless treatment of steroid hormone bacterial residues. Background Art

[0002] At present, more than 400 steroid drugs have been produced globally, ranking the second largest category of chemical drugs after antibiotics. China is a major producer of steroid raw materials, with the annual output of steroid drugs accounting for about 1 / 3 of the world's total output. Among them, the production capacity and actual output of corticosteroid raw materials both rank first in the world.

[0003] As a steroid hormone, androstenedione can be produced through a microbial transformation process. Limited by the separation and extraction efficiency, androstenedione in the fermentation broth cannot be completely extracted, resulting in inevitable residues of a small amount of androstenedione and other metabolites in the bacterial residues, which may affect the reproductive, immune, and nervous systems of various organisms and have certain ecological toxicity. At the same time, the bacterial residues produced during the microbial transformation process also contain rich mycelium and residual organic matter, sugars, proteins, minerals and other nutrient components, which are a resource with broad development prospects.

[0004] The traditional treatment method for hormone bacterial residues is incineration, which not only eliminates the residues after production but also wastes the relevant resources in the residues. Compared with the traditional treatment method for hormone bacterial residues, anaerobic digestion is an effective way to realize the resource utilization of organic waste, which can degrade various solid organic waste and high-concentration organic wastewater, etc., and at the same time can achieve a win-win situation for energy and ecology. And anaerobic fermentation has a lower requirement for the C / N ratio of raw materials, and does not require adding a large amount of straw-like auxiliary materials to adjust the C / N ratio and moisture content in the treatment of bacterial residues, reducing the regional limitation of technology application.

[0005] In the anaerobic treatment of organic waste, volatile fatty acids are often produced, but under the tolerance conditions of methanogens, the volatile fatty acids in the bacterial residues will be converted into biogas.

[0006] Therefore, it is of great significance to develop a method for harmless resource utilization of androstenedione bacterial residues. Summary of the Invention

[0007] Object of the Invention: The object of the present invention is to provide a method for harmless treatment of steroid hormone bacterial residues. While reducing the quantity and harmlessly treating the steroid hormone bacterial residues, the present invention accumulates biogas production for resource utilization.

[0008] Technical Solution: The object of the present invention is achieved by the following technical solutions:

[0009] The present invention provides a method for harmless treatment of steroid hormone bacterial residues, comprising the following steps:

[0010] (1) Use the androstenedione-containing bacterial residue raw material as a substrate and mix it with anaerobic activated sludge;

[0011] The mass ratio of the androstenedione-containing bacterial residue raw material to the anaerobic activated sludge is 1:3 to 1:1;

[0012] (2) At a fermentation temperature of 35-40 °C or 50-60 °C, in an inert atmosphere, conduct sealed anaerobic fermentation for 28-32 days to collect biogas. At the same time, detect the concentration of the steroid hormone androstenedione.

[0013] A preferred embodiment of the present invention is that in step (1), by thermally pre-treating the androstenedione-containing bacterial residue raw material, the macromolecular organic matter that is difficult to biodegrade in the bacterial residue can be converted into easily degradable small-molecule substances, improving the available organic matter in the raw material, such as soluble sugars, volatile fatty acids, soluble proteins, and amino acids.

[0014] Further preferably, the temperature of the thermal pre-treatment is 80-160 °C and the time is 30-90 min.

[0015] Preferably, in step (1), the concentration of androstenedione in the androstenedione-containing bacterial residue raw material is 20-40 mg / L.

[0016] Preferably, in step (1), the water content of the androstenedione-containing bacterial residue raw material is 95-99%, and the proportion of volatile solids in the androstenedione-containing bacterial residue raw material to the total solids is 60-70%.

[0017] Preferably, in step (1), the androstenedione bacterial residue is a single substrate with a C / N ratio of 7.8.

[0018] A preferred embodiment of the present invention is that in step (1), androstenedione pure product with a concentration of 0-10 g / L is added externally to the androstenedione-containing bacterial residue raw material.

[0019] Further preferably, androstenedione pure product with a concentration of 1-8 g / L is added externally to the androstenedione-containing bacterial residue raw material.

[0020] Even more preferably, androstenedione pure product with a concentration of 8 g / L is added externally to the androstenedione-containing bacterial residue raw material.

[0021] Preferably, in step (2), the fermentation temperature is 35-40 °C.

[0022] In the present invention, the fermentation process is placed in a constant temperature water bath or a constant temperature incubator to maintain a constant temperature.

[0023] Preferably, in step (2), an inert atmosphere is formed by the method of blowing nitrogen to remove oxygen.

[0024] Preferably, in step (2), the anaerobic fermentation is carried out in a semi - continuous or batch mode.

[0025] Preferably, in step (2), the cycle of anaerobic fermentation is 28 days until the daily gas production approaches zero.

[0026] Beneficial effects:

[0027] 1. The present invention adopts anaerobic fermentation technology, provides a high - value resource utilization technology for efficient treatment of androstenedione bacterial residue, can reduce the quantity and harmlessize the bacterial residue, eliminate the residual harmful substance androstenedione, and accumulate biogas for resource utilization.

[0028] 2. By controlling the fermentation temperature and time and removing the dissolved oxygen in the material by nitrogen blowing, the present invention enables the conversion of volatile fatty acids in the material into methane, realizing the accumulation of biogas. The present invention uses androstenedione bacterial residue with a C / N ratio of 7.8 as a single substrate, without the need for co - fermentation with high - carbon - containing organic matter, saving costs, having a continuous effect, and being easy to control fermentation conditions. Description of the drawings

[0029] Figure 1 Biogas production rate of androstenedione bacterial residue under medium - temperature (35 °C) and high - temperature (55 °C) conditions;

[0030] Figure 2 Biogas production rate of androstenedione bacterial residue under different mass ratios in Example 3 and Example 4;

[0031] Figure 3 Biogas production rate of androstenedione bacterial residue before heat pretreatment in Example 3 and after heat pretreatment in Example 5 - 6;

[0032] Figure 4 Removal rate of volatile total solids VS in Example 1 - 6;

[0033] Figure 5 Removal rate of androstenedione in Example 1 - 6;

[0034] Figure 6 Protein degradation rate in Example 3, Example 5, and Example 6;

[0035] Figure 7 Fat degradation rate in Example 3, Example 5, and Example 6;

[0036] Figure 8 Biogas production rate of androstenedione bacterial residue under different androstenedione contents in Example 9 and Example 10;

[0037] Figure 9 Daily removal rate and cumulative removal rate of androstenedione under different androstenedione contents in Example 9 and Example 10;

[0038] Figure 10 The biogas production rate of androstenedione bacterial residue after degradation by degrading bacteria. Specific embodiments

[0039] The technical solution of the present invention will be described in detail below through specific embodiments, but the protection scope of the present invention is not limited to the described embodiments.

[0040] The androstenedione bacterial residue raw material used in the examples and comparative examples of the present invention is androstenedione fermentation residue (ADR) from: Jiangsu Bio Environmental Protection Technology Co., Ltd., Jiangsu. The moisture content of this raw material is 96.88%, the androstenedione content is 38.15 mg / L, the proportion of volatile solids in the androstenedione bacterial residue raw material in the total solids is 67.13%, and at the same time, the raw material contains 8.54 g / L of protein and 11.08 g / L of fat.

[0041] The anaerobic activated sludge used in the examples and comparative examples of the present invention is derived from anaerobic digestion sludge collected from a continuously stirred tank reactor that has been operating for a long time at Changzhou University.

[0042] Pure androstenedione: Source: Shanghai Macklin Biochemical Technology Co., Ltd.; Purity: 98%.

[0043] Example 1

[0044] Take 0.33 g of androstenedione bacterial residue raw material and 1 g of anaerobic activated sludge, so that the mass ratio of the raw material to the sludge is 1:3. Add deionized water to a working volume of 100 ml, pour it into a serum bottle, blow N2, seal it with a butyl rubber stopper and an aluminum cap, and place it in a constant temperature water bath at 35 °C for 32 days to ferment, and collect biogas.

[0045] Example 2

[0046] Take 0.33 g of androstenedione bacterial residue raw material and 1 g of anaerobic activated sludge, so that the mass ratio of the raw material to the sludge is 1:3. Add deionized water to a working volume of 100 ml, pour it into a serum bottle, blow N2, seal it with a butyl rubber stopper and an aluminum cap, and place it in a constant temperature water bath at 55 °C for 32 days to ferment, and collect biogas.

[0047] Figure 1 The biogas production rate of androstenedione bacterial residue under medium temperature (35 °C) and high temperature (55 °C) conditions.

[0048] It can be seen from the figure that after 32 days of anaerobic fermentation, under the condition of medium temperature of 35 °C, the biogas production rate of androstenedione bacterial residue is 0.692 g-COD / g-VS; under the condition of high temperature of 55 °C, the biogas production rate of androstenedione bacterial residue is 0.369 g-COD / g-VS. Therefore, the medium temperature condition is more suitable for the anaerobic fermentation of androstenedione bacterial residue.

[0049] Example 3

[0050] Take 3.806 g of androstenedione-containing bacterial residue raw material and 3.806 g of anaerobic activated sludge, so that the mass ratio of the raw material to the sludge is 1:1. Add deionized water to a working volume of 500 ml, pour it into a reactor, blow N2, seal it with a butyl rubber stopper and an aluminum cover, and place it in a 35 °C constant temperature water bath for fermentation for 28 days to collect biogas.

[0051] Example 4

[0052] Take 3.806 g of androstenedione-containing bacterial residue raw material and 11.418 g of anaerobic activated sludge, so that the mass ratio of the raw material to the sludge is 1:3. Add deionized water to a working volume of 500 ml, pour it into a reactor, blow N2, seal it with a butyl rubber stopper and an aluminum cover, and place it in a 35 °C constant temperature water bath for fermentation for 28 days to collect biogas.

[0053] Figure 2 The biogas production rate of androstenedione-containing bacterial residue under different mass ratio conditions.

[0054] It can be seen from the figure that after 28 days of anaerobic fermentation, the biogas production rate of androstenedione-containing bacterial residue with an inoculation ratio of 1:3 is significantly higher than that of androstenedione-containing bacterial residue with an inoculation ratio of 1:1.

[0055] Example 5

[0056] Place the raw material in a high-temperature pressure-resistant bottle and conduct heat pretreatment at 80 °C in an oil bath for 90 min. Take 3.806 g of the heat-pretreated androstenedione-containing bacterial residue raw material and 3.806 g of anaerobic activated sludge, so that the mass ratio of the raw material to the sludge is 1:1. Add deionized water to a working volume of 500 ml, pour it into a reactor, blow N2, seal it with a butyl rubber stopper and an aluminum cover, and place it in a 35 °C constant temperature water bath for fermentation for 28 days to collect biogas.

[0057] Example 6

[0058] Place the raw material in a high-temperature pressure-resistant bottle and conduct heat pretreatment at 160 °C in an oil bath for 30 min. Take 3.806 g of the heat-pretreated androstenedione-containing bacterial residue raw material and 3.806 g of anaerobic activated sludge, so that the mass ratio of the raw material to the sludge is 1:1. Add deionized water to a working volume of 500 ml, pour it into a reactor, blow N2, seal it with a butyl rubber stopper and an aluminum cover, and place it in a 35 °C constant temperature water bath for fermentation for 28 days to collect biogas.

[0059] Figure 3 The biogas production rate of androstenedione-containing bacterial residue without heat pretreatment in Example 3 and after heat pretreatment in Examples 5 - 6.

[0060] It can be seen from the figure that after 28 days of anaerobic fermentation, under the conditions of 80 °C and 90 min (Example 5), the biogas production rate of androstenedione bacterial residue is significantly higher than that of androstenedione bacterial residue without heat treatment (Example 3) and under the conditions of 160 °C and 30 min (Example 6).

[0061] Example 7

[0062] The present invention measured the reduction and harmlessness. The removal rates of volatile total solids content (VS) and androstenedione corresponding to Examples 1-6 are as Figure 4 、 Figure 5 。

[0063] Figure 4 are the removal rates of VS for Examples 1-6;

[0064] Figure 5 are the removal rates of androstenedione for Examples 1-6.

[0065] It can be seen from the figure that under the conditions of 80 °C and 90 min, the removal rates of VS and androstenedione of the bacterial residue reach the highest, which are 54.2% and 98.6%.

[0066] Example 8

[0067] The present invention measured the protein and fat before and after fermentation in Examples 3, 5, and 6. The protein degradation rates and fat degradation rates corresponding to Examples 3, 5-6 are as Figure 6 、 Figure 7 。

[0068] Figure 6 are the protein degradation rates;

[0069] Figure 7 are the fat degradation rates.

[0070] It can be seen from the figure that under the conditions of 160 °C and 30 min, the protein degradation rate and fat degradation rate of the bacterial residue reach the highest, which are 61.7% and 83.4%.

[0071] Example 9

[0072] Take 0.5 g of androstenedione bacterial residue raw material and 1 g of anaerobic activated sludge, so that the mass ratio of the raw material to the sludge is 1:2. Add deionized water to a working volume of 100 ml, pour it into a serum bottle, blow N2, seal it with a butyl rubber stopper and an aluminum cap, and place it in a 35 °C constant temperature water bath for fermentation for 28 days to collect biogas.

[0073] Example 10

[0074] Take 0.5 g of the androstenedione-containing bacterial residue raw material and 1 g of anaerobic activated sludge, so that the mass ratio of the raw material to the sludge is 1:2. Before fermentation, add 0.2 mL of an androstenedione pure product solution with a concentration of 8 g / L prepared with methanol (99% purity) as an organic solvent to the androstenedione-containing bacterial residue raw material, and then mix it with the anaerobic activated sludge;

[0075] Add deionized water to a working volume of 100 ml, pour it into a serum bottle, blow N2, seal it with a butyl rubber stopper and an aluminum cap, and place it in a 35°C constant temperature water bath for fermentation for 28 days to collect biogas.

[0076] Figure 8 It is the biogas production rate of the androstenedione-containing bacterial residue under different androstenedione contents.

[0077] Figure 9 It is the single-day removal rate and cumulative removal rate of androstenedione under different androstenedione contents.

[0078] It can be seen from the figure that after 28 days of anaerobic fermentation, compared with the bacterial residue without additional androstenedione (Example 9), the biogas production rate of the bacterial residue with exogenous androstenedione added (Example 10) is higher, indicating that the higher the content of androstenedione in the anaerobic fermentation process, the higher the biogas production rate. The highest single-day removal rate of androstenedione in the exogenous added bacterial residue during anaerobic fermentation is 17.15%, which is higher than the highest single-day removal rate of 12.19% without addition. The higher the androstenedione content, the higher the removal rate of androstenedione during anaerobic fermentation.

[0079] Control Example 1

[0080] Inoculate Burkholderia metalica from the soil near Changzhou University into the androstenedione-containing bacterial residue raw material, and completely degrade androstenedione after culturing at 37°C and 180 rpm for 30 h.

[0081] Take 0.5 g of the bacterial residue raw material degraded by Burkholderia metalica, 1 g of anaerobic activated sludge, so that the mass ratio of the raw material to the sludge is 1:2, add deionized water to a working volume of 100 ml, pour it into a serum bottle, blow N2, seal it with a butyl rubber stopper and an aluminum cap, and place it in a 35°C constant temperature water bath for fermentation for 28 days to collect biogas.

[0082] Figure 10 It is the biogas production rate of the androstenedione-containing bacterial residue after degradation by the degrading bacteria.

[0083] It can be seen from the figure that after 28 days of anaerobic fermentation, compared with the bacterial residue without treating androstenedione (as it is, Example 9), the biogas production rate of the bacterial residue with androstenedione degraded by Burkholderia metalica is lower, indicating that androstenedione has an impact on the biogas production rate during anaerobic fermentation.

[0084] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as a limitation on the present invention itself. Various changes may be made in its form and details without departing from the spirit and scope of the present invention as defined by the appended claims.

Claims

1. A method for harmless treatment of steroid hormone bacterial residue, characterized in that, It includes the following steps: (1) Using the androstenedione-containing bacterial residue raw material as a substrate, mix it with anaerobic activated sludge; The mass ratio of the androstenedione-containing bacterial residue raw material to the anaerobic activated sludge is 1:3 to 1:1; (2) At a fermentation temperature of 35 - 40 °C or 50 - 60 °C, in an inert atmosphere, conduct sealed anaerobic fermentation for 28 - 32 days to collect biogas.

2. The method according to claim 1, wherein In step (1), the androstenedione-containing bacterial residue raw material is subjected to thermal pretreatment.

3. The method according to claim 2, wherein The temperature of the thermal pretreatment is 80 - 160 °C, and the time is 30 - 90 min.

4. The method according to claim 1, characterized in that, In step (1), the concentration of androstenedione in the androstenedione-containing bacterial residue raw material is 20 - 40 mg / L.

5. The method according to claim 1, characterized in that, In step (1), the moisture content of the androstenedione-containing bacterial residue raw material is 95 - 99%, and the proportion of volatile solids in the androstenedione-containing bacterial residue raw material in the total solids is 60 - 70%.

6. The method according to claim 1, wherein In step (1), the androstenedione bacterial residue is a single substrate with a C / N ratio of 7.

8.

7. The method according to claim 1, wherein In step (1), androstenedione pure product with a concentration of 0 - 10 g / L is externally added to the androstenedione-containing bacterial residue raw material.

8. The method according to claim 1, wherein In step (2), the fermentation temperature is 35 - 40 °C.

9. The method according to claim 1, characterized in that, In step (2), an inert atmosphere is formed by the method of blowing nitrogen to remove oxygen.

10. The method according to claim 1, characterized in that, In step (2), the cycle of anaerobic fermentation is 28 days.

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