CN110777090B - A kind of food waste treatment bacteria and its application - Google Patents

Abstract

The invention relates to the technical field of environmental microorganisms, in particular to a kitchen waste treatment bacterial strain and its application. The present invention is a kind of Franconibacter helveticus FH-JW1, the preservation number is CCTCC M 2019649; it also provides a liquid microbial agent and a solid microbial agent containing Franconibacter helveticus FH-JW1 and preparation thereof The method; the third aspect provides the application of a bacterial agent containing Franconibacter helveticus FH-JW1 in kitchen waste and perishable organic waste. The food waste treatment bacteria of the invention is resistant to high temperature, can rapidly degrade food waste and other perishable organic wastes under high temperature conditions, and realizes rapid and efficient biological reduction and stabilization of perishable organic wastes such as food waste.

Description

A kind of food waste treatment bacteria and its application

technical field

The invention relates to the technical field of environmental microorganisms, in particular to a kitchen waste treatment bacterial strain and its application.

Background technique

With the continuous advancement of my country's urbanization process and the improvement of people's living standards, the amount of domestic waste generated in my country has increased year by year. In 2017, the annual removal and transportation of urban domestic waste in my country exceeded 200 million tons, of which the amount of kitchen waste generated was 99.72 million tons, equivalent to 270,000 tons per day. At present, 46 cities in my country have started garbage classification. By 2020, all cities will fully implement the garbage classification system. Among them, the proportion of classified kitchen waste is relatively large, accounting for about 50% of domestic waste. The garbage has high moisture content, low calorific value, high organic matter content, and is easily perishable and smelly. If it is not treated effectively, it will easily cause the reproduction of pathogenic bacteria, the breeding of mosquitoes and flies and water pollution, and also bring huge pressure on the living environment. Therefore, The effective treatment of kitchen waste is an important part of urban ecological environment management.

At present, the commonly used kitchen waste treatment technologies at home and abroad include dry incineration, sanitary landfill, ecological feed, anaerobic digestion, and aerobic composting. At present, most of the cities in my country are still dealing with kitchen waste by traditional incineration and landfill, which cannot be used as a resource. It is a huge waste of kitchen waste and brings a heavy burden to local finances. Even in cities (such as Beijing and Shanghai) that vigorously develop kitchen waste recycling technology, the proportion of recycling is relatively low. In recent years, with the deepening of domestic waste classification, the amount of kitchen waste has risen sharply. Due to the increasing tension of land resources and the continuous enhancement of people's awareness of environmental protection, the amount of food waste in landfill will be less and less. At the same time, due to the high moisture content and low calorific value of kitchen waste, the cost of incineration treatment has increased significantly. Therefore, aerobic biological treatment technology with the core of reduction and resource utilization has received extensive attention. Aerobic bioconversion treatment has broad market application prospects due to its advantages of large processing capacity, short time, no secondary pollutants from biogas slurry and biogas residue, and the ability to produce organic fertilizers or soil conditioners. However, traditional aerobic biotransformation still faces problems such as low biotransformation rate and long fermentation cycle, which urgently need technological breakthroughs to solve.

SUMMARY OF THE INVENTION

In view of the above-mentioned shortcomings of the prior art, the purpose of the present invention is to provide a kitchen waste treatment strain and its application, which are used to solve the problems in the prior art.

In order to realize the above-mentioned purpose and other related purposes, the present invention provides a first aspect to provide a kind of Franconibacter helveticus, which is preserved in CCTCC, China Type Culture Collection Center, and the strain name is Franconibacter helveticus FH-JW1, and the preservation date is 2019.8.19, the deposit number is CCTCC M2019649.

The Franconibacter helveticus FH-JW1 contains the gene sequence shown in SEQ ID NO.1.

The Franconibacter helveticus FH-JW1 strain was treated for 48 hours, and the wet weight loss rate of food and beverage waste was over 92%.

A second aspect of the present invention provides a liquid bacterial agent, the liquid bacterial agent at least comprises Franconibacter helveticus FH-JW1.

The third aspect of the present invention provides a preparation method of Franconibacter helveticus FH-JW1 liquid bacterial agent.

The preparation method at least comprises the following steps:

1) Inoculate pure bacteria in 10mL liquid medium, and cultivate at 50℃ and 180r/min for 12-16h;

2) Inoculation amount of 1% (volume) was inserted into the same liquid medium for expansion culture, and cultured at 50°C and 180r/min for 16-20h.

The liquid culture medium is composed of: peptone 3 g/L, yeast extract 2 g/L, NaCl 3 g/L, MgCl ₂ 0.125 g/L, CaCl ₂ 0.5 g/L, FeSO ₄ 7H ₂ O 0.01 g/L, Trace element solution (Na ₂ MoO ₄ 12g/L, VOSO ₄ ·xH ₂ O 1g/L, MnCl ₂ 5g/L, ZnSO ₄ ·7H ₂ O 0.6g/L, CuSO ₄ ·5H ₂ O 0.15g/L, CoCl ₂ 6H ₂ O 8g/L, NiCl ₂ 6H ₂ O 0.2g/L) 100 μL/L, and the pH value was adjusted to 7-7.5.

A fourth aspect of the present invention provides a solid inoculum, the solid inoculum at least comprises a liquid inoculum of Franconibacter helveticus FH-JW1 and a carrier.

The carrier is one or more selected from sawdust, rice husk and wheat bran.

A fifth aspect of the present invention provides a method for preparing a solid inoculum, the method at least comprising mixing and drying a liquid inoculum and a carrier.

Preferably, the mass ratio of the carrier to the liquid bacterial agent is 1:0.5-1:3.

The sixth aspect of the present invention provides the application of Franconibacter helveticus FH-JW1 in the treatment of kitchen waste.

Preferably, the food waste can be one or more of food waste and kitchen waste.

A seventh aspect of the present invention provides an application of Franconibacter helveticus FH-JW1 in the treatment of perishable organic garbage, which at least includes the following steps:

1) Sort the perishable organic waste to be treated to remove sundries;

2) Add the sorted perishable organic waste into the Franconibacter helveticus FH-JW1 solid inoculum, and the mass ratio of the solid inoculum to the kitchen waste is 1:1 to 1:5;

3) Stir the mixture evenly, and carry out aerobic fermentation at 40-70°C, with continuous ventilation and stirring during the process.

Preferably, the perishable organic waste is selected from one or more of catering waste, kitchen waste and vegetable farm waste.

Dispose of food waste according to the steps of the seventh aspect, the mass ratio of solid bacterial agent to food waste is 1:3, the aerobic fermentation temperature is set to 60°C, and the wet weight loss rate of food waste is calculated according to formula ①, and calculated according to formula ② Dry weight loss rate of food waste. The results showed that the wet weight loss rate was 92% at 48h, and the dry weight loss rate was 57.6% at 24h.

Wet weight loss rate (%) = (m _{initial wet} – m _{final wet} )/m _{material wet} × 100% ①

Among them, m _{initial wetness} is the total humidity of the solid bacteria agent and garbage at the initial time, m _{final wetness} is the total humidity of the solid bacteria agent and the garbage at the time of detection, m is the total humidity of the garbage, unit: g.

Dry weight loss rate (%) = (m _{dry at the beginning} – m at the _end )/m _dry × 100% ②

Among them, m _{starting dry} is the total dry amount of solid bacteria agent and garbage at the initial time, m _{final dry} amount is the total dry amount of solid bacteria agent and garbage at the time of detection, m _{material dry} is the total dry amount of garbage, unit: g. The dry weight was measured by the constant weight method at 105°C.

As mentioned above, the food waste treatment bacteria of the present invention has the following beneficial effects:

1) High temperature resistance, can quickly degrade kitchen waste under high temperature conditions of 40-70 °C;

2) Under high temperature conditions, it has the ability to produce protease and amylase, and can efficiently degrade organic substances such as protein and starch in food waste. When the temperature is 60 °C, the reduction rate of food waste reaches 92%;

3) Rapid and efficient biological reduction and stabilization of kitchen waste can be achieved.

Description of drawings

Fig. 1 shows the schematic diagram of the phylogenetic tree of the strain FH-JW1 of the present invention;

Fig. 2 shows the wet weight reduction rate of strain FH-JW1 of the present invention to food and beverage waste;

Fig. 3 shows the dry weight reduction rate of strain FH-JW1 of the present invention to food and beverage waste;

Figure 4 shows the wet weight loss rate of the strain FH-JW1 of the present invention on mixed perishable waste.

Detailed ways

The embodiments of the present invention are described below through specific specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

Before further describing the specific embodiments of the present invention, it should be understood that the protection scope of the present invention is not limited to the following specific specific embodiments; it should also be understood that the terms used in the examples of the present invention are for describing specific specific embodiments, Rather than limiting the scope of the invention; in the specification and claims of the present invention, the singular forms "a", "an" and "the" include the plural forms unless the context clearly dictates otherwise.

When numerical ranges are given in the examples, it is to be understood that, unless otherwise indicated herein, both endpoints of each numerical range and any number between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by one of ordinary skill in the art. In addition to the specific methods, equipment and materials used in the embodiments, according to the mastery of the prior art by those skilled in the art and the description of the present invention, the methods, equipment and materials described in the embodiments of the present invention can also be used Any methods, devices and materials similar or equivalent to those of the prior art can be used to implement the present invention.

Unless otherwise specified, the experimental methods, detection methods and preparation methods disclosed in the present invention all adopt the conventional molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology and related fields in the technical field. conventional technology. These techniques are well described in the existing literature.

The present invention obtains a bacterial strain capable of efficiently degrading kitchen waste, named FH-JW1, through preliminary screening of the separation medium, re-screening of the enzyme-producing medium, and high-temperature aerobic biological treatment of kitchen waste. The taxonomic status of the strain was identified as Franconibacter helveticus by comprehensive analysis of the data of colony and cell morphology, physiological and biochemical characteristics, and 16S rRNA gene sequence determination. The strain was deposited in the China Center for Type Culture Collection (CCTCC) with a preservation date of 2019.8.19, a preservation number of CCTCC M 2019649, and a preservation address at the Chinese Type Culture Collection Center of Wuhan University, Bayi Road, Wuchang District, Wuhan City, Hubei Province.

The main biological characteristics of the bacterial species are Gram-negative bacteria, rod-shaped, and the colony shape is transparent and round, with neat edges and smooth and moist surfaces.

The experimental results of the bacteria degrading kitchen waste show that the strain can improve the reduction rate of kitchen waste under aerobic conditions, achieve the purpose of reducing and stabilizing kitchen waste, and can be used to degrade perishable organic waste. For example, catering waste, kitchen waste, and vegetable yard waste have high application value.

In the following examples, the media used are as follows:

Separation and purification medium: peptone 3g/L, yeast extract 2g/L, NaCl 3g/L, MgCl ₂ 0.125g/L, CaCl ₂ 0.5g/L, FeSO ₄ 7H ₂ O 0.01g/L, trace element solution (Na ₂ MoO ₄ 12g/L, VOSO ₄ ·xH ₂ O 1g/L, MnCl ₂ 5g/L, ZnSO ₄ ·7H ₂ O 0.6g/L, CuSO ₄ ·5H ₂ O 0.15g/L, CoCl ₂ 6H ₂ O 8g/L, NiCl ₂ 6H ₂ O 0.2g/L) 100μL/L, agar 20g/L, and the pH value was adjusted to 7-7.5.

Liquid medium: peptone 3g/L, yeast extract 2g/L, NaCl 3g/L, MgCl ₂ 0.125g/L, CaCl ₂ 0.5g/L, FeSO ₄ 7H ₂ O 0.01g/L, trace element solution ( Na ₂ MoO ₄ 12g/L, VOSO ₄ ·xH ₂ O 1g/L, MnCl ₂ 5g/L, ZnSO ₄ ·7H ₂ O 0.6g/L, CuSO ₄ ·5H ₂ O 0.15g/L, CoCl ₂ 6H ₂ O 8g/L, NiCl ₂ 6H ₂ O 0.2g/L) 100 μL/L, and the pH value was adjusted to 7-7.5.

Protease screening medium: casein 10g/L, peptone 3g/L, yeast extract 2g/L, NaCl 3g/L, MgCl ₂ 0.125g/L, CaCl ₂ 0.5g/L, FeSO ₄ 7H ₂ O 0.01g /L, trace element solution (Na ₂ MoO ₄ 12g/L, VOSO ₄ ·xH ₂ O 1g/L, MnCl ₂ 5g/L, ZnSO ₄ ·7H ₂ O 0.6g/L, CuSO ₄ ·5H ₂ O 0.15g /L, CoCl ₂ 6H ₂ O 8g/L, NiCl ₂ 6H ₂ O 0.2g/L) 100 μL/L, agar 20g/L, and the pH value was adjusted to 7-7.5.

Amylase screening medium: soluble starch 10g/L, peptone 3g/L, yeast extract 2g/L, NaCl 3g/L, MgCl ₂ 0.125g/L, CaCl ₂ 0.5g/L, FeSO ₄ 7H ₂ O 0.01 g/L, trace element solution (Na ₂ MoO ₄ 12g/L, VOSO ₄ ·xH ₂ O 1g/L, MnCl ₂ 5g/L, ZnSO ₄ ·7H ₂ O 0.6g/L, CuSO ₄ ·5H ₂ O 0.15 g/L, CoCl ₂ 6H ₂ O 8g/L, NiCl ₂ 6H ₂ O 0.2g/L) 100 μL/L, agar 20 g/L, pH value adjusted to 7-7.5.

Example 1: Isolation, screening and performance determination of strain FH-JW1

The separation and purification process of the strain FH-JW1 of the present invention: the test sample is collected from a fermenter in a kitchen waste treatment plant, and after adding sterile water and mixing, gradient dilution is performed, and 200 μL of the dilutions of different gradients are applied to the separation and purification culture. After culturing in a constant temperature incubator at 60°C for 24 hours, single colonies with different shapes and sizes were picked, streaked and purified, and numbered for preservation. A total of 17 strains were obtained through primary screening.

The pure strains obtained from the primary screening were inoculated onto the protease screening medium and the amylase screening medium by spot-joining, and the enzyme-producing ability of the strains was measured, so as to carry out re-screening. After culturing at 60 °C for 48 h, the colony diameter (d) and the diameter of the transparent circle (D) were measured, and the D/d was calculated, and the strains with strong protein and starch degradation abilities were selected.

The strains obtained by re-screening were inoculated into liquid medium, cultured at 50°C and 180r/min for 16h, centrifuged 20mL of bacterial liquid, washed with sterile saline, resuspended to 10mL, attached to 30g of sawdust, and inoculated into 70g Preliminary verification was carried out in the kitchen waste, and the wet weight reduction rate was calculated according to formula ①. The results showed that the wet weight reduction rate of the kitchen waste inoculated with the strain FH-JW1 was the largest, reaching 92% at 48h.

Wet weight loss rate (%) = (m _{initial wet} – m _{final wet} )/m _{material wet} × 100% ①

Among them, m _{initial wetness} is the total moisture content of wood chips and garbage at the initial time, m _{final wetness} is the total moisture content of wood chips and garbage at the time of detection, m is the total moisture content of garbage, unit: g.

The 16S rDNA sequence of the strain is shown in SEQ ID NO: 1, and the 16S rRNA sequence was submitted at NCBI, and the homology sequence alignment analysis was performed with the software and GenBank, and the MEGA 6 software was used to construct a phylogenetic tree of the strain (as shown in Figure 1 ). shown). Based on the above information, the strain FH-JW1 was identified as Franconibacter helveticus.

Example 2: Preparation of inoculum of Francoella swiss

Preparation of liquid inoculum: inoculate FH-JW1 pure bacteria in 10mL liquid medium, cultivate in a constant temperature shaker at 50°C and 180r/min for 12-16h, and then inoculate it with 1% (volume) inoculum. The liquid inoculum of the strain can be obtained by inserting it into the same liquid medium to expand the culture, and culture at 50°C and 180r/min for 16-20h.

Preparation of solid inoculum: the liquid inoculum of the strain is prepared by mixing and drying carriers such as sawdust, rice husk, wheat bran, etc. Preferably, the mass ratio of the carrier to the liquid inoculum is 1:0.5～1:3 .

Example 3: Treatment effect of strain FH-JW1 on food waste

Sort the food waste to be treated, remove debris such as bones, plastics, chopsticks, etc., add Francoella swiss solid inoculum to the sorted food waste, and the mass ratio of solid inoculum to food waste is 1 : 3. After stirring evenly, carry out aerobic fermentation at 60 °C. During the process, uninterrupted ventilation and stirring are required to ensure uniform heating and under aerobic conditions. The treatment system was weighed every 12 hours to obtain the reduction rate of food and beverage waste. The dry weight and wet weight reduction effects are shown in Figures 2 and 3.

It can be seen from Figure 2 that the experimental group with access to the strain FH-JW1 showed a higher reduction rate than the control group without access to the strain, and the wet weight loss rate of the access strain reached 92% at 48h. %, an increase of 23.0% compared with 74.8% of the control group. Further from the dry weight loss rate in Figure 3, it can be seen that the dry weight loss rate of the food waste by inserting Francoella swissis almost reached the highest value of 57.6% in 24 hours, while the maximum dry weight loss rate of the control group was 57.6%. only reached 2.3%. It can be seen that the access to Francoella swiss FH-JW1 can significantly improve the reduction effect of food waste, and achieve rapid biological reduction and stabilization of food waste.

Example 4: Treatment effect of strain FH-JW1 on mixed perishable organic waste

The three kinds of perishable organic wastes to be treated, such as food waste, kitchen waste and vegetable waste, are mixed in a mass ratio of 1:1:1, and then sorted to remove bones, plastics, chopsticks and other debris. The solid bacteria agent of Francoella swiss is added to the mixed perishable organic waste. The mass ratio of the solid bacteria agent to the mixed perishable organic waste is 1:2. After stirring evenly, aerobic fermentation is carried out at 70 °C. Uninterrupted ventilation and stirring to ensure uniform heating and under aerobic conditions. The treatment system was weighed every 12 hours to obtain the wet weight reduction rate of the mixed perishable organic waste. The reduction effect is shown in Figure 4. The wet weight loss rate of the inoculated strain FH-JW1 was as high as 94% within 48h, and approached the maximum value after 24h, while the wet weight loss rate of the control group without inoculation was as high as 78.1%. Compared with the control group, the strain FH-JW1 increased the wet weight loss rate of mixed perishable organic waste by 20.4%. It can be seen that under aerobic conditions, Francoella swiss can not only improve the reduction rate of single garbage, but also can be used to degrade mixed perishable organic garbage such as food waste, kitchen waste, vegetable waste, etc., which has a high application. value and broad application prospects.

To sum up, the present invention effectively overcomes various shortcomings in the prior art and has high industrial utilization value.

The above-mentioned embodiments merely illustrate the principles and effects of the present invention, but are not intended to limit the present invention. Anyone skilled in the art can modify or change the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or changes made by those with ordinary knowledge in the technical field without departing from the spirit and technical idea disclosed in the present invention should still be covered by the claims of the present invention.

sequence listing

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<120> A kind of food waste treatment bacteria and its application

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Claims (12)

1. Francisella helveticusFranconibacter helveticusFH-JW1 with a preservation number of CCTCC M2019649.

2. The bacteria of claim 1, selected from the group consisting of FFranconibacter helveticusFH-JW1, characterized in that the Shigella verruckeriFranconibacter helveticusFH-JW1 contains the gene sequence shown in SEQ ID NO. 1.

3. A liquid microbial agent comprising at least the bacteria of the genus Buddha of claim 1 or 2Franconibacter helveticusFH-JW1 or its progeny culture obtained microbial population.

4. A method for preparing a liquid microbial inoculum according to claim 3, which comprises at least the following steps:

1) inoculating the pure bacteria into 10mL of liquid culture medium, and culturing for 12-16 h;

2) inoculating the strain into a liquid culture medium with the inoculation amount of 1% for amplification culture for 16-20 h.

5. A solid microbial inoculum, which comprises at least a carrier and the liquid microbial inoculum according to claim 3.

6. The solid microbial inoculum according to claim 5, wherein the carrier is selected from one or more of wood chips, rice hulls and wheat bran.

7. A method for preparing the solid microbial inoculum according to claim 5 or 6, which comprises at least the following steps:

1) mixing the carrier and the liquid microbial inoculum according to the mass ratio of 1: 0.5-1: 3;

2) drying the mixture of step 1).

8. A strain of the bacterium of claim 1 or 2, of the genus Francisella helveticusFranconibacter helveticusFH-JW1 is applied to the treatment of kitchen waste.

9. The application of claim 8, wherein the kitchen waste is at least one or more of food waste and kitchen waste.

10. A strain of the bacterium of claim 1 or 2, of the genus Francisella helveticusFranconibacter helveticusApplication of FH-JW1 in the treatment of perishable organic waste.

11. The use according to claim 10, wherein the perishable organic waste is at least one or more of food waste, kitchen waste, and vegetable waste.

12. Use according to claim 10, characterized in that it comprises at least the following steps:

1) sorting perishable organic garbage to be treated to remove impurities;

2) adding the sorted perishable organic garbage into the solid microbial inoculum according to claim 5 or 6, wherein the mass ratio of the solid microbial inoculum to the garbage is 1: 1-1: 5, and stirring;

3) carrying out aerobic fermentation at 40-70 ℃, and continuously ventilating and stirring in the process.

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