Preparation method of microbial deodorant as well as product and application of microbial deodorant

Abstract

The invention relates to a preparation method of a microbial deodorant as well as a product and application of the microbial deodorant, the preparation method comprises the following steps: inoculating a seed solution of a bacillus coagulans BC66 strain into a fermentation culture medium for primary culture to obtain a primary fermentation solution; and inoculating a seed solution of a lactobacillus acidophilus LA71 strain into the primary fermentation solution, and carrying out secondary culture to obtain the microbial deodorant. The brand-new microbial deodorant with the excellent deodorization effect is developed, the BC66 strain and the LA71 strain are combined and sequentially fermented to obtain fermentation liquor, the two probiotics can produce organic acid such as lactic acid and butyric acid through fermentation substrates, meanwhile, main odor components such as ammonia gas and hydrogen sulfide can be effectively adsorbed and decomposed, and the deodorization effect is good. In addition, the deodorant also has an excellent inhibition effect on pathogenic bacteria and putrefying bacteria, the two probiotics are mutually matched and have synergistic interaction on the effects, and the prepared deodorant is green, safe, environment-friendly and free of secondary pollution, and is beneficial for enterprises to realize green production.

Description

Technical Field

[0001] The present invention belongs to the field of microbial agents, and relates to a preparation method, product and application of a microbial deodorant. Background Art

[0002] With the improvement of China's aquaculture industry and economic level, the pollution of air environment caused by the landfill odor and incineration tail gas generated by the odor of aquaculture and the increase in per capita domestic waste is becoming increasingly serious. Compared with the traditional chemical deodorization method, the microbial deodorization method has the advantages of being more green and safe, having no secondary pollution, lasting effect, and low cost. It has become an inevitable trend to gradually weaken or even replace the chemical deodorization method by combining biological deodorization (especially microbial deodorization) with physical methods.

[0003] In addition, with the steady development of China's economy, probiotics have witnessed rapid growth in the field of big health. While meeting the growing material needs of the people, it also brings a large amount of environmental protection pressure, that is, the treatment pressure of fermentation waste liquid (the supernatant collected after centrifugation and other steps of the bacterial sludge in the fermentation broth, hereinafter collectively referred to as the supernatant). Usually, there are still a small number of live bacteria in the supernatant, and it also contains the metabolites of the strains during the growth process, such as organic acids, vitamins, antibiotics, growth promoting factors, etc. If directly discharged into the sewage treatment system, due to its high concentration and complex organic components, the treatment cost is relatively high, and the beneficial metabolites will also be wasted. Therefore, if the supernatant can be recycled, it can not only effectively solve the waste liquid discharge pressure of probiotic production enterprises, but also bring additional economic benefits and achieve green production of enterprises. Summary of the Invention

[0004] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a preparation method, product and application of a microbial deodorant.

[0005] To achieve the purpose of this invention, the following technical solutions are adopted:

[0006] In the first aspect, the present invention provides a preparation method of a microbial deodorant, and the preparation method includes the following steps: inoculating the seed liquid of Bacillus coagulans into a fermentation medium for primary culture to obtain a primary fermentation broth; then inoculating the seed liquid of Lactobacillus acidophilus into the primary fermentation broth for secondary culture to obtain the microbial deodorant;

[0007] The Bacillus coagulans is Bacillus coagulans BC66 strain with the preservation number of CGMCC 21879;

[0008] The Lactobacillus acidophilus is Lactobacillus acidophilus LA71 strain with the preservation number of CGMCC 21765.

[0009] The present invention has developed a brand-new microbial deodorant with excellent deodorizing effect, which is a fermentation broth obtained by sequentially fermenting BC66 strain and LA71 strain. Both probiotics can utilize fermentation substrates to produce organic acids such as lactic acid and butyric acid, and at the same time can effectively adsorb and decompose main odor components such as ammonia and hydrogen sulfide. In addition, they also have excellent inhibitory effects on pathogenic bacteria and spoilage bacteria. The two probiotics cooperate with each other and synergistically enhance the efficacy in the above-mentioned effects. The prepared deodorant is green, safe, environmentally friendly and has no secondary pollution, which helps enterprises to achieve green production.

[0010] Preferably, the viable count of BC66 strain in the microbial deodorant is not less than 1×10 9 CFU / mL, such as 1×10 9 CFU / mL, 5×10 9 CFU / mL, 8×10 9 CFU / mL, 1×10 10 CFU / mL, 5×10 10 CFU / mL, etc.; the viable count of LA71 strain is not less than 1×10 8 CFU / mL, such as 1×10 8 CFU / mL, 5×10 8 CFU / mL, 1×10 9 CFU / mL, 5×10 9 CFU / mL, 8×10 9 CFU / mL, 1×10 10 CFU / mL, 5×10 10 CFU / mL, etc.; other specific point values within this numerical range can be selected and will not be elaborated one by one here.

[0011] Preferably, the primary culture is carried out at 30-38°C (such as 30°C, 32°C, 33°C, 34°C, 35°C, 36°C, 38°C, etc.) and a rotation speed of 100-300 rpm (such as 100 rpm, 150 rpm, 200 rpm, 250 rpm, 300 rpm, etc.) for 24-48 h (such as 24 h, 26 h, 30 h, 34 h, 36 h, 40 h, 48 h, etc.). Other specific point values within the above-mentioned numerical ranges can be selected and will not be elaborated one by one here.

[0012] Preferably, the secondary culture is carried out at 30 - 38 °C (such as 30 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 38 °C, etc.) and a rotation speed of 100 - 300 rpm (such as 100 rpm, 150 rpm, 200 rpm, 250 rpm, 300 rpm, etc.) for 60 - 84 h (such as 60 h, 64 h, 68 h, 72 h, 76 h, 80 h, 84 h, etc.). Other specific point values within the above numerical ranges can be selected and will not be elaborated here one by one.

[0013] The end point of the secondary culture is that the pH value of the fermentation broth is stable below 4.8, such as pH = 4.7, pH = 4.6, pH = 4.5, pH = 4.4, pH = 4.3, pH = 4.2, etc.

[0014] Preferably, the viable count in the seed solution of Bacillus coagulans and the seed solution of Lactobacillus acidophilus is not less than 1×10 9 CFU / mL, such as 1×10 9 CFU / mL, 5×10 9 CFU / mL, 8×10 9 CFU / mL, 1×10 10 CFU / mL, 5×10 10 CFU / mL, etc.; the inoculation amount is 1 - 5%, such as 1%, 2%, 3%, 4%, 5%, etc.; other specific point values within the above numerical ranges can be selected and will not be elaborated here one by one.

[0015] Preferably, the fermentation medium includes the supernatant waste liquid of probiotic fermentation.

[0016] Preferably, the probiotics include any one or a combination of at least two of Bacillus coagulans, Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium animalis, Pediococcus acidilactici, Lactobacillus casei / paracasei, Lactobacillus buchneri, or Lactobacillus reuteri.

[0017] The preparation method of the microbial deodorant involved in the present invention can use the supernatant waste liquid of probiotic fermentation as the fermentation substrate, realizing the resource utilization of the waste liquid of bacterial fermentation. It not only reduces the discharge of the waste liquid of bacterial fermentation and reduces the environmental protection pressure of the enterprise, but also realizes the resource utilization, turning waste into treasure, and finding a reasonable benefit growth point for the enterprise.

[0018] Preferably, the fermentation medium further includes a supplementary carbon source and / or a supplementary antioxidant.

[0019] Preferably, the supplementary carbon source includes glucose and / or molasses.

[0020] Preferably, the supplementary antioxidant includes complex vitamins and / or potassium sorbate.

[0021] In a second aspect, the present invention provides a microbial deodorant prepared by the preparation method according to the first aspect.

[0022] In a third aspect, the present invention provides the use of the microbial deodorant according to the second aspect in removing ammonia and hydrogen sulfide in the spaces of livestock and poultry farms, toilets, and garbage transfer stations.

[0023] Preferably, the microbial deodorant is used by direct spraying or spraying after dilution with water.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The present invention has developed a brand-new microbial deodorant with excellent deodorizing effect. It is a fermentation broth obtained by sequentially co-fermenting BC66 strain and LA71 strain. Both probiotic bacteria can utilize the fermentation substrate to produce organic acids such as lactic acid and butyric acid, and at the same time can effectively adsorb and decompose the main odor components such as ammonia and hydrogen sulfide. In addition, it has an excellent inhibitory effect on pathogenic bacteria and spoilage bacteria. The two probiotic bacteria cooperate with each other and synergistically enhance the effect in the above-mentioned effects. The prepared deodorant is green, safe, environmentally friendly, and has no secondary pollution, which helps enterprises achieve green production.

[0026] The preparation method of the microbial deodorant involved in the present invention can use the supernatant waste liquid of probiotic fermentation as the fermentation substrate, realizing the resource utilization of the waste liquid of bacterial fermentation. It not only reduces the discharge of the waste liquid of bacterial fermentation and reduces the environmental protection pressure of enterprises, but also realizes resource utilization, turning waste into treasure, and finding a reasonable benefit growth point for enterprises.

[0027] The taxonomic name of the BC66 strain involved in the present invention is Bacillus coagulans, and it is preserved in the China General Microbiological Culture Collection Center. The preservation time is March 8, 2021, and the preservation number is CGMCC 21879. The address is: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0028] The taxonomic name of the LA71 strain involved in the present invention is Lactobacillus acidophilus, and it is preserved in the China General Microbiological Culture Collection Center. The preservation time is January 29, 2021, and the preservation number is CGMCC 21765. The address is: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Detailed Embodiments

[0029] To further illustrate the technical means and effects adopted by the present invention, the following further describes the technical solutions of the present invention in combination with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0030] The classification and naming of the BC66 strain used in the following examples is Bacillus coagulans. The preservation time is March 8, 2021, and the preservation number is CGMCC 21879.

[0031] The classification and naming of the LA71 strain used in the following examples is Lactobacillus acidophilus. The preservation time is January 29, 2021, and the preservation number is CGMCC 21765.

[0032] The compound vitamins involved in the following examples were purchased from Shenzhen Love Biotechnology Co., Ltd.; the molasses was purchased from Guangxi Huashang Guotang Trading Co., Ltd.

[0033] The probiotic fermentation supernatant liquids used in the following Examples 1-3 are the supernatant waste liquids obtained by centrifuging Lactobacillus casei after fermentation at 37°C for 72 hours; the supernatant waste liquids obtained by centrifuging Lactobacillus plantarum after fermentation at 37°C for 72 hours; the supernatant waste liquids obtained by centrifuging Lactobacillus acidophilus after fermentation at 37°C for 72 hours.

[0034] Example 1

[0035] This example provides a microbial deodorant, and its preparation method is as follows:

[0036] (1) Inoculate the Bacillus coagulans BC66 strain and the Lactobacillus acidophilus LA71 strain into LB liquid medium and MRS liquid medium respectively, and perform shake flask culture at 35°C and 200 r / min for 18 h to obtain 800 mL of seed liquid for each of the two strains, and their effective viable counts are 3.8×10 9 CFU / mL and 2.4×10 9 CFU / mL respectively;

[0037] (2) Transfer 80 L of the probiotic fermentation supernatant waste liquid into a 100 L fermentation tank, and add 1.6 kg of molasses, 800 g of compound vitamins, and 800 g of potassium sorbate respectively and mix evenly as the fermentation medium;

[0038] (3) Inoculate 800 mL of the Bacillus coagulans BC66 seed liquid into the above fermentation medium, control the temperature at 35°C and the rotation speed at 200 r / min, and culture for 36 h to obtain a primary fermentation liquid; then inoculate 800 mL of the Lactobacillus acidophilus LA71 seed liquid into the primary fermentation liquid, control the temperature at 35°C and the rotation speed at 200 r / min, and after culturing for 72 h, the pH of the fermentation liquid is stabilized at about 4.5 to obtain the microbial deodorant.

[0039] After detection, the effective viable count of Bacillus coagulans BC66 in the microbial deodorant is 5.3×109 CFU / mL, the effective viable count of Lactobacillus acidophilus LA71 is 4.6×10 8 CFU / mL.

[0040] Example 2

[0041] This example provides a microbial deodorant, and its preparation method is as follows:

[0042] (1) Inoculate Bacillus coagulans BC66 strain and Lactobacillus acidophilus LA71 strain into LB liquid medium and MRS liquid medium respectively, and perform shake flask culture at 35°C and 200 r / min for 18 h to obtain 1000 mL of seed liquid for each of the two strains, and their effective viable counts are 3.8×10 9 CFU / mL and 2.4×10 9 CFU / mL;

[0043] (2) Transfer 80 L of the supernatant waste liquid of probiotic fermentation into a fermentation tank with a specification of 100 L, add 1.6 kg of molasses and 800 g of potassium sorbate, and mix evenly as the fermentation medium;

[0044] (3) Inoculate 1000 mL of Bacillus coagulans BC66 seed liquid into the above fermentation medium, control the temperature at 37, the rotation speed at 300 r / min, and culture for 48 h to obtain the primary fermentation liquid; then inoculate 1000 mL of Lactobacillus acidophilus LA71 seed liquid into the primary fermentation liquid, control the temperature at 37°C, the rotation speed at 300 r / min, and culture for 64 h until the pH of the fermentation liquid stabilizes at about 4.7 to obtain the microbial deodorant.

[0045] After detection, the effective viable count of Bacillus coagulans BC66 in the microbial deodorant is 6.3×10 9 CFU / mL, and the effective viable count of Lactobacillus acidophilus LA71 is 2.5×10 8 CFU / mL.

[0046] Example 3

[0047] This example provides a microbial deodorant, and its preparation method is as follows:

[0048] (1) Inoculate Bacillus coagulans BC66 strain and Lactobacillus acidophilus LA71 strain into LB liquid medium and MRS liquid medium respectively, and perform shake flask culture at 35°C and 200 r / min for 18 h to obtain 1500 mL of seed liquid for each of the two strains, and their effective viable counts are 3.8×10 9 CFU / mL and 2.4×10 9 CFU / mL;

[0049] (2) Transfer 80 L of the supernatant waste liquid of probiotic fermentation into a fermenter with a specification of 100 L, and add 1.6 kg of molasses and 800 g of compound vitamins respectively, and mix evenly as the fermentation medium;

[0050] (3) Inoculate 1500 mL of the Bacillus coagulans BC66 seed liquid into the above fermentation medium, control the temperature at 32 °C, the rotation speed at 100 r / min, and culture for 30 h to obtain the primary fermentation liquid; then inoculate 1500 mL of the Lactobacillus acidophilus LA71 seed liquid into the primary fermentation liquid, control the temperature at 32 °C, the rotation speed at 100 r / min, and after culturing for 80 h, the pH of the fermentation liquid is stable at about 4.3 to obtain the microbial deodorant.

[0051] After detection, the effective viable count of Bacillus coagulans BC66 in the microbial deodorant is 5.8×10 9 CFU / mL, and the effective viable count of Lactobacillus acidophilus LA71 is 5.5×10 8 CFU / mL.

[0052] Comparative Example 1

[0053] This comparative example provides a microbial deodorant, and the difference in its preparation method from Example 1 is only that the Bacillus coagulans BC66 strain is replaced by the Bacillus coagulans ATCC7050 strain, and all culture conditions remain unchanged.

[0054] After detection, the effective viable count of Bacillus coagulans ATCC7050 in the microbial deodorant is 6.9×10 9 CFU / mL, and the effective viable count of Lactobacillus acidophilus LA71 is 5.2×10 8 CFU / mL.

[0055] Comparative Example 2

[0056] This comparative example provides a microbial deodorant, and the difference in its preparation method from Example 1 is only that the Lactobacillus acidophilus LA71 strain is replaced by the Lactobacillus acidophilus ATCC4356 strain, and all culture conditions remain unchanged.

[0057] After detection, the effective viable count of Lactobacillus acidophilus ATCC4356 in the microbial deodorant is 7.2×10 9 CFU / mL, and the effective viable count of Lactobacillus acidophilus LA71 is 5.0×10 8 CFU / mL.

[0058] Comparative Example 3

[0059] This comparative example provides a microbial deodorant, and the difference in its preparation method from Example 1 is only that only the BC66 strain is fermented:

[0060] (1) Inoculate Bacillus coagulans BC66 strain into LB liquid medium and culture it in a shaking flask at 35°C and 200 r / min for 18 h to obtain the seed liquid of the strain, and its effective viable count is 3.8×10 9 CFU / mL;

[0061] (2) Transfer 80 L of the supernatant waste liquid of probiotic fermentation into a 100-L fermenter, and add 1.6 kg of molasses, 800 g of compound vitamins, and 800 g of potassium sorbate and mix evenly as the fermentation medium;

[0062] (3) Inoculate 1600 mL of the Bacillus coagulans BC66 seed liquid into the above fermentation medium, control the temperature at 35°C and the rotation speed at 200 r / min, and culture for 36 h to obtain the microbial deodorant.

[0063] After detection, the effective viable count of Bacillus coagulans BC66 in the microbial deodorant is 6.7×10 9 CFU / mL.

[0064] Comparative Example 4

[0065] This comparative example provides a microbial deodorant, and the difference in its preparation method from that of Example 1 is only that only LA71 strain fermentation is carried out:

[0066] (1) Inoculate Lactobacillus acidophilus LA71 strain into MRS liquid medium and culture it in a shaking flask at 35°C and 200 r / min for 18 h to obtain the seed liquid of the strain, and its effective viable count is 2.4×10 9 CFU / mL;

[0067] (2) Transfer 80 L of the supernatant waste liquid of probiotic fermentation into a 100-L fermenter, and add 1.6 kg of molasses, 800 g of compound vitamins, and 800 g of potassium sorbate and mix evenly as the fermentation medium;

[0068] (3) Inoculate 1600 mL of the Lactobacillus acidophilus LA71 seed liquid into the above fermentation medium, control the temperature at 35°C and the rotation speed at 200 r / min, and culture for 72 h to obtain the microbial deodorant.

[0069] After detection, the effective viable count of Lactobacillus acidophilus LA71 in the microbial deodorant is 6.1×10 9 CFU / mL.

[0070] Test Example

[0071] The microbial deodorants prepared in Examples 1-3 and Comparative Examples 1-4 were applied to different chicken coops in a certain chicken farm (the environmental conditions of each chicken coop had no obvious differences). The microbial deodorant was diluted 80 times with water and atomized and sprayed in the chicken coop once in the morning and once in the afternoon, with a dosage of 80 mL / m after each dilution. 3 After 48 hours of spraying, sensory evaluation was carried out and a pump suction type gas detector was used to detect the concentrations of ammonia and hydrogen sulfide, and the removal rates of ammonia and hydrogen sulfide were calculated. The results are shown in Table 1.

[0072] Table 1

[0073] Group Sensory effect Ammonia removal rate (%) Hydrogen sulfide removal rate (%) Example 1 Odorless 98.7％ 86.3％ Example 2 Odorless 97.2％ 83.7％ Example 3 Odorless 95.8％ 82.6％ Comparative Example 1 Occasional odor 88.9％ 78.2％ Comparative Example 2 Occasional odor 87.5％ 76.7％ Comparative Example 3 Occasional odor 85.0％ 76.5％ Comparative Example 4 Occasional odor 88.4％ 70.1％

[0074] It can be seen from the data results in Table 1 that the microbial deodorant involved in the present invention has excellent effects on removing the main odor components ammonia and hydrogen sulfide, and the BC66 strain and the LA71 strain have obvious synergistic effects in the above-mentioned effects.

[0075] The applicant declares that the technical scheme of the present invention is illustrated by the above-mentioned examples, but the present invention is not limited to the above-mentioned examples, that is, it does not mean that the present invention must rely on the above-mentioned examples to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent replacement of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc. all fall within the protection scope and the disclosure scope of the present invention.

[0076] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept scope of the present invention, various simple modifications can be made to the technical scheme of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0077] In addition, it should be noted that, in the case of no contradiction, the various specific technical features described in the above specific embodiments can be combined in any suitable manner. In order to avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. A preparation method of a microbial deodorant, characterized in that, The preparation method comprises the following steps: inoculating the seed liquid of Bacillus coagulans into a fermentation medium for primary culture to obtain a primary fermentation broth; then inoculating the seed liquid of Lactobacillus acidophilus into the primary fermentation broth for secondary culture to obtain the microbial deodorant; The Bacillus coagulans is Bacillus coagulans BC66 strain with the preservation number of CGMCC 21879; The Lactobacillus acidophilus is Lactobacillus acidophilus LA71 strain with the preservation number of CGMCC 21765.

2. The preparation method of the microbial deodorant according to claim 1, characterized in that, The viable count of strain BC66 in the microbial deodorant is not less than 1×10 9 CFU / mL; the viable count of strain LA71 is not less than 1×10 8 CFU / mL.

3. The preparation method of the microbial deodorant according to claim 1, characterized in that, The primary culture is carried out at 30-38 °C and a rotation speed of 100-300 rpm for 24-48 h.

4. The preparation method of the microbial deodorant according to claim 1, characterized in that, The secondary culture is carried out at 30-38 °C and a rotation speed of 100-300 rpm for 60-84 h; The end point of the secondary culture is that the pH value of the fermentation broth is stable below 4.

8.

5. The preparation method of the microbial deodorant according to claim 1, characterized in that, The viable cell counts in the seed solution of Bacillus coagulans and the seed solution of Lactobacillus acidophilus are not less than 1×10 9 CFU / mL; the inoculation amounts are both 1-5%.

6. The preparation method of the microbial deodorant according to claim 1, characterized in that The fermentation medium comprises the supernatant waste liquid of probiotic fermentation; Preferably, the probiotic comprises any one or a combination of at least two of Bacillus coagulans, Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium animalis, Pediococcus acidilactici, Lactobacillus casei / paracasei, Lactobacillus buchneri or Lactobacillus reuteri.

7. The preparation method of the microbial deodorant according to claim 1, characterized in that, The fermentation medium further comprises a supplementary carbon source and / or a supplementary antioxidant; Preferably, the supplementary carbon source comprises glucose and / or molasses; Preferably, the supplementary antioxidant comprises a compound vitamin and / or potassium sorbate.

8. A microbial deodorant prepared by the preparation method according to any one of claims 1-7.

9. Use of the microbial deodorant according to claim 8 in removing ammonia and hydrogen sulfide in the spaces of livestock and poultry farms, toilets and garbage transfer stations.

10. The application according to claim 9, wherein, The microbial deodorant is directly sprayed or sprayed after being diluted with water.