CN113980933A - Preparation of complex enzyme and method for treating wastewater sludge by using complex enzyme - Google Patents
Preparation of complex enzyme and method for treating wastewater sludge by using complex enzyme Download PDFInfo
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- 239000002351 wastewater Substances 0.000 title claims abstract description 103
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 66
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 66
- 238000000034 method Methods 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title abstract description 6
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- 238000000855 fermentation Methods 0.000 claims abstract description 24
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- 230000000694 effects Effects 0.000 claims abstract description 14
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- 150000001875 compounds Chemical class 0.000 claims abstract description 11
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- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 claims description 9
- 102100022624 Glucoamylase Human genes 0.000 claims description 9
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F11/00—Treatment of sludge; Devices therefor
- C02F11/02—Biological treatment
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2428—Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
- C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
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- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01003—Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel
Abstract
The invention discloses a preparation method of complex enzyme and a method for treating wastewater and sludge by using the complex enzyme, and belongs to the field of solid waste treatment and disposal. The method comprises the processes of preparing a compound enzyme fermentation substrate, preparing a compound enzyme, hydrolyzing waste water sludge by the compound enzyme and performing anaerobic fermentation on the waste water sludge, wherein the waste water sludge is used as a raw material to prepare a cheap high-activity compound enzyme for the ultra-fast hydrolysis of the waste water sludge, and a hydrolysate is quickly converted into methane through anaerobic digestion. The complex enzyme prepared by the method hydrolyzes wastewater sludge, the COD concentration dissolved by hydrolysate is improved, further the methane yield is greatly improved, and the method has the advantages of low cost, short reaction period, small occupied area and less residue discharge.
Description
Technical Field
The invention relates to the field of solid waste treatment and disposal, in particular to preparation of a complex enzyme and a method for treating wastewater and sludge by using the complex enzyme.
Background
With the growth of population and rapid development of economy in China, the yield of wastewater and sludge is increased sharply, and the treatment situation is increasingly severe. At present, incineration and landfill are the main disposal means for wastewater and sludge. However, incineration energy consumption is high, toxic and harmful gases are easily generated, and incineration ash needs to be disposed of in landfill, and the land resources currently used for landfill are extremely limited, so incineration and landfill are not an environment-friendly waste water and sludge disposal method. In addition to wastewater sludge, high energy consumption is another challenge facing current sewage treatment plants. In 2018, the total electricity consumption of sewage treatment plants in the whole country is about 169 hundred million kilowatt hours, and the total amount of sludge generated is about 680 million tons (dry weight).
The anaerobic digestion and methane (energy) recovery is considered to be an efficient waste water sludge resource utilization approach due to the simple and efficient process characteristics, the clean and environment-friendly technical idea and the outstanding contribution to energy conservation and emission reduction. However, since the composition of the wastewater sludge is extremely complex and the hydrolysis efficiency is low, only about 30% of the organic matter can be converted into methane, and the remaining wastewater sludge still needs to be incinerated and disposed of in landfills. Proper pretreatment can greatly promote the hydrolysis of wastewater sludge, shorten the anaerobic fermentation time, improve the methane yield and reduce the discharge of solid residues. Some pretreatment methods such as hydrothermal treatment and chemical treatment have been used to improve the hydrolysis efficiency of wastewater sludge and methane yield. Li Wei utilizes a hydrothermal method to pretreat wastewater sludge, so that the concentration of dissolved organic matters in the wastewater sludge is increased, and finally the yield of methane is increased by more than 2 times. However, the hydrothermal pretreatment method has high energy consumption and is easy to generate byproducts which are difficult to degrade, so that the whole anaerobic digestion reaction period still needs about 30 days. The Fang Wei utilizes chemical alkali to pretreat the wastewater sludge, the dissolved COD concentration of the pretreated wastewater sludge is obviously increased, and finally the methane yield is improved by nearly 60 percent. However, chemical pretreatment is too costly due to the continuous addition of chemicals and is prone to secondary pollution.
Enzymatic pretreatment is considered to be a green and environmentally friendly pretreatment method that can be used to promote hydrolysis of sludge and increase its gas production efficiency. Compared with the traditional hydrothermal treatment and chemical pretreatment, the enzyme pretreatment has the advantages of low energy consumption, no chemical agent addition, no generation of refractory substances, simple operation and the like. Currently, several commercial enzymes have been used to increase the efficiency of hydrolysis of wastewater sludge. Wilson Parawira respectively pretreats the sludge by using commercial cellulase and beta-glucosidase, and the methane yield is respectively improved by 12% and 15% in the anaerobic digestion process. However, commercial enzyme pretreatment is still limited by factors such as excessive cost, and commercial enzymes are generally present in a single form, which makes it difficult to achieve efficient hydrolysis of complex wastewater sludge. In addition to the above bottleneck, another problem of the current anaerobic digestion technology for wastewater and sludge is that a large amount of biogas residues is generated after the wastewater and sludge is subjected to anaerobic digestion, and the biogas residues subjected to anaerobic digestion are difficult to apply to agriculture due to the existence of pathogenic bacteria and heavy metals, and still need to be subjected to subsequent incineration or landfill disposal.
In a word, the problems of long reaction period, low pretreatment efficiency, high cost, difficult biogas residue disposal and the like exist in the anaerobic digestion of the waste water sludge at present, and the large-scale industrialization of the anaerobic digestion technology of the waste water sludge is restricted.
Disclosure of Invention
Aiming at the problems of long reaction period, low pretreatment efficiency, high cost, easy generation of secondary pollution, difficult residue treatment and the like in the existing anaerobic digestion technology of wastewater sludge, the invention provides a preparation method of a complex enzyme and a method for treating wastewater sludge by using the complex enzyme.
The invention is realized by the following technical scheme.
On the one hand, the invention provides a method for treating wastewater sludge by using complex enzyme, which comprises the following steps:
removing stones and plastic impurities in the wastewater sludge, and screening the wastewater sludge after impurity removal;
sterilizing part of screened wastewater sludge, inoculating Aspergillus oryzae bacteria liquid into the sterilized wastewater sludge, and culturing at a fermentation temperature to obtain a complex enzyme;
the weight ratio of the dry weight to the mass is (3-6): 100, mixing the complex enzyme with the screened non-sterilized wastewater sludge, and carrying out heating hydrolysis reaction to obtain a hydrolysate;
taking anaerobic sludge, and mixing the hydrolysate with the inoculum anaerobic sludge according to the mass ratio of volatile solids of 1: (0.5-3) adding the mixture into a reactor, and stirring and fermenting at a constant temperature under anaerobic fermentation temperature to obtain a fermentation product methane.
Preferably, the wastewater sludge is manually sorted, and then the sorted wastewater sludge is screened by a 80-mesh screen.
Preferably, the screened portion of the wastewater sludge is sterilized at 121 ℃ for 15 minutes.
Preferably, 0.05-0.15mL of Aspergillus oryzae liquid is inoculated to the sterilized part of the waste water sludge per gram of dry weight of the sludge, i.e. 0.5-1.5X 10 of the waste water sludge per gram of dry weight of the sludge is inoculated6An Aspergillus oryzae fungal spore.
Preferably, the complex enzyme fermentation temperature is 30 ℃, the oxygen content is 21-60%, the humidity is 50% -80%, the original pH of the waste water sludge is 7.5, and the culture is carried out for 8-12 days.
Preferably, the hydrolysis reaction time is 8-16 hours, the hydrolysis temperature is 40-70 ℃, and the stirring speed is 0-200 rpm/min.
Preferably, the anaerobic fermentation temperature is 37 ℃, the reaction time is 5-10 days, and the stirring speed is 0-200 rpm.
On the other hand, the invention provides the compound enzyme for treating wastewater and sludge prepared by the method.
In the complex enzyme prepared by the invention, the activity of glucoamylase is not lower than 37(U/g dry weight), and the activity of protease is not lower than 41(U/g dry weight); the compound enzyme hydrolyzes the wastewater sludge, and the concentration of dissolved COD is not lower than 39.7g/L (namely, no less than 397 g of dissolved COD is generated per kg of dry sludge); the anaerobic fermentation of the wastewater sludge can ensure that the methane yield is not lower than 323mL/g VS.
Due to the adoption of the technical scheme, the invention has the following beneficial effects:
in the scheme, the high-activity compound enzyme is prepared by using the waste water sludge as a raw material, is rich in glucoamylase, protease, amylase and cellulase, can realize rapid hydrolysis of the waste water sludge with complex components, overcomes the defects of high energy consumption, easy generation of inhibiting substances, secondary pollution and the like of the traditional waste water sludge hydrothermal and chemical pretreatment, and also solves the problems of single component, high cost and low hydrolysis efficiency of commercial enzymes. In addition, compared with the traditional anaerobic digestion technology of wastewater and sludge, the method can simultaneously realize the improvement of the methane yield by 2.3 times and the solid reduction by 68.5 percent, has the advantages of low cost, short reaction period, small occupied area, less residue discharge and the like, and is a wastewater and sludge treatment mode with very promising prospect.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the invention and together with the description serve to explain the principles of the invention:
FIG. 1 is a block diagram of the process of the present invention.
Detailed Description
The present invention will now be described in detail with reference to the drawings and specific embodiments, wherein the exemplary embodiments and descriptions of the present invention are provided to explain the present invention without limiting the invention thereto.
As shown in FIG. 1, the embodiment of the invention provides a method for treating wastewater sludge by using complex enzyme, which comprises the following steps:
1) preparing a complex enzyme fermentation substrate:
collected wastewater sludge is manually sorted and screened by a 80-mesh screen, and 10g (dry weight) of screened wastewater sludge is taken and put into a plurality of 500ml conical flasks for sterilization and standby. The sterilization temperature and time were 121 ℃ and 15 minutes, respectively.
2) Preparing a complex enzyme:
inoculating Aspergillus oryzae solution into the sterilized wastewater sludge at an inoculation ratio of 5-15%, and inoculating 0.05-0.15mL of Aspergillus oryzae solution per gram of dry weight into the wastewater sludge, i.e. inoculating 0.5-1.5 × 10 per gram of dry weight into the wastewater sludge6The Aspergillus oryzae fungal spores are put into an incubator to produce the complex enzyme, the incubation time is 8-12 days, the incubation temperature is 30 ℃, the oxygen content is 21-60%, the humidity is 50% -80%, the original pH of the wastewater sludge is 7.5, and the water content is 90%.
3) The compound enzyme hydrolyzes the wastewater sludge:
according to the mass ratio of (3-6): 100 mixing the complex enzyme with the screened non-sterilized wastewater sludge, and carrying out heating hydrolysis reaction for 8-16 hours at the hydrolysis temperature of 40-70 ℃ and the stirring speed of 0-200rpm to obtain a hydrolysate.
4) Anaerobic fermentation of wastewater sludge:
taking anaerobic sludge, and mixing the hydrolysate with the inoculum anaerobic sludge according to the mass ratio of volatile solids of 1: (0.5-3), adding the mixture into a reactor, carrying out anaerobic fermentation at the culture temperature of 37 ℃ and the stirring speed of 0-200rpm, stirring and fermenting at constant temperature, and reacting for 5-10 days to obtain a fermentation product, namely the biomethane.
The invention is further illustrated by the following different examples.
1) Preparing a complex enzyme fermentation substrate:
collected wastewater sludge is manually sorted and screened by a 80-mesh screen, and 10g (dry weight) of screened wastewater sludge is taken and put into a plurality of 500ml conical flasks for sterilization and standby. The sterilization temperature is 121 ℃, and the sterilization time is 15 minutes.
2) Preparing a complex enzyme: examples 1-9 are examples of complex enzyme preparation.
3) The compound enzyme hydrolyzes the wastewater sludge: examples 10-18 and comparative example 1 are examples of complex enzyme hydrolysis of wastewater sludge.
4) Anaerobic fermentation of wastewater sludge:
examples 19-25 are wastewater sludge anaerobic fermentation examples.
Example 1
Inoculating Aspergillus oryzae solution into the sterilized wastewater sludge at an inoculation ratio of 10%, and inoculating 0.1mL of Aspergillus oryzae solution into the wastewater sludge per gram of dry weight (i.e. inoculating 1.0 × 10 per gram of dry weight of wastewater sludge with 1.0 × 10 per gram of dry weight of wastewater sludge)6Aspergillus oryzae fungal spores) is put into an incubator to produce complex enzyme, the culture time is 10 days, the culture temperature is 30 ℃, the oxygen content is 40%, the humidity is 50%, the original pH of the wastewater sludge is 7.5, and the water content is 90%. The glucoamylase and protease activity of the complex enzyme was measured at this point to be 57 and 76U/g (dry weight), respectively.
Examples 2 to 9
The conditions and results are shown in the following table, and the other conditions and steps are the same as in example 1
As can be seen from the above examples, the activity of glucoamylase in the prepared complex enzyme is not less than 37(U/g dry weight), and the activity of protease is not less than 41(U/g dry weight).
Example 10
5g of the prepared complex enzyme (dry weight, namely 285U of glucoamylase activity and 380U of protease activity) and 100g (dry weight) of the screened unsterilized wastewater sludge are put into a 2L reactor for hydrolysis, the hydrolysis time is 12 hours, the hydrolysis temperature is 60 ℃, and the stirring speed is 100 rpm. The dissolved COD concentration of the final hydrolysate was 65.7 g/L.
Comparative example 1
Commercial glucoamylase with the same enzyme activity, protease and 100g (dry weight) of screened wastewater sludge were put into a 2L reactor for hydrolysis at 60 ℃ for 12 hours and 100 rpm. The rest of the procedure was the same as in example 10. The resulting hydrolyzate had a dissolved COD concentration of 62.3 g/L.
As can be seen from example 10 and comparative example 1, the complex enzyme prepared from wastewater sludge and developed by the invention has better hydrolysis effect of wastewater sludge than commercial glucoamylase and protease, because the complex enzyme self-made from wastewater sludge and developed by the invention contains 11U/g amylase and 13U/g cellulase besides 57U/g glucoamylase and 76U/g protease, which shows that the complex enzyme self-made from wastewater sludge and commercial enzyme is more suitable for hydrolyzing wastewater sludge with complex components, and the pretreatment cost is reduced.
Examples 11 to 18
The conditions and results are shown in the following table, and the other conditions and steps are the same as those of example 10
As can be seen from the above examples, the self-made complex enzyme is used for hydrolyzing the wastewater sludge, the concentration of dissolved COD is not lower than 39.7g/L (namely, no less than 397 g of dissolved COD is generated per kilogram of dry sludge), and the complex enzyme prepared by the method has excellent hydrolysis effect for treating the wastewater sludge.
Example 19
Taking 100ml of the obtained hydrolysate as a substrate for preparing the biological methane, wherein the fermentation temperature is 37 ℃, the fermentation time is 7 days under the anaerobic condition, the stirring speed is 100rpm, and the adding ratio (VS/VS) of the hydrolysate to the anaerobic sludge is 1: 2. The final methane yield obtained was 514mL/g VS.
Examples 20 to 25
The conditions and results are shown in the following table, and the other conditions and steps are the same as in example 19
From the above examples, it can be seen that the yield of the prepared methane is not lower than 323(mL/g VS) after the wastewater sludge is pretreated by the self-made complex enzyme, and the wastewater sludge treated by the self-made complex enzyme has excellent methane production potential.
The present invention is not limited to the above embodiments, and based on the technical solutions disclosed in the present invention, those skilled in the art can make some substitutions and modifications to some technical features without creative efforts, and these substitutions and modifications are all within the protection scope of the present invention.
Claims (9)
1. A method for treating wastewater sludge by using complex enzyme is characterized by comprising the following steps:
removing stones and plastic impurities in the wastewater sludge, and screening the wastewater sludge after impurity removal;
sterilizing part of screened wastewater sludge, inoculating Aspergillus oryzae bacteria liquid into the sterilized wastewater sludge, and culturing at a fermentation temperature to obtain a complex enzyme;
the weight ratio of the dry weight to the mass is (3-6): 100, mixing the complex enzyme with the screened non-sterilized wastewater sludge, and carrying out heating hydrolysis reaction to obtain a hydrolysate;
taking anaerobic sludge, and mixing the hydrolysate with the inoculum anaerobic sludge according to the mass ratio of volatile solids of 1: (0.5-3) adding the mixture into a reactor, and stirring and fermenting at a constant temperature under anaerobic fermentation temperature to obtain a fermentation product methane.
2. The method for treating wastewater sludge by using the complex enzyme as claimed in claim 1, wherein the wastewater sludge is manually sorted, and then the sorted wastewater sludge is screened by an 80-mesh screen.
3. The method for treating wastewater sludge by using complex enzyme as claimed in claim 1, wherein the screened part of wastewater sludge is sterilized for 15 minutes at 121 ℃.
4. The method for treating wastewater sludge by using compound enzyme as claimed in claim 1, wherein 0.05-0.15mL of Aspergillus oryzae solution per gram of dry weight of sludge is inoculated into the sterilized part of wastewater sludge.
5. The method for treating wastewater sludge by using the complex enzyme as claimed in claim 1, wherein the fermentation temperature of the complex enzyme is 30 ℃, the oxygen content is 21-60%, the humidity is 50% -80%, the original pH of the wastewater sludge is 7.5, and the wastewater sludge is cultured for 8-12 days.
6. The method for treating wastewater sludge by using the complex enzyme as claimed in claim 1, wherein the hydrolysis reaction time is 8-16 hours, the hydrolysis temperature is 40-70 ℃, and the stirring speed is 0-200 rpm.
7. The method for treating wastewater sludge by using the complex enzyme as claimed in claim 1, wherein the anaerobic fermentation temperature is 37 ℃, the reaction time is 5-10 days, and the stirring speed is 0-200 rpm/min.
8. A complex enzyme for treating wastewater sludge prepared by the method of any one of claims 1-7.
9. The complex enzyme of claim 8, wherein the activity of glucoamylase in the complex enzyme is not less than 37U/g dry weight, and the activity of protease is not less than 41U/g dry weight; the compound enzyme hydrolyzes the wastewater sludge, and each kilogram of dry sludge generates no less than 397 grams of dissolved COD; the anaerobic fermentation of the wastewater sludge can ensure that the methane yield is not lower than 323mL/g VS.
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