CN108251324B - Bacillus subtilis, microbial inoculum containing bacillus subtilis, application of microbial inoculum, method for degrading vomitoxin and kit - Google Patents
Bacillus subtilis, microbial inoculum containing bacillus subtilis, application of microbial inoculum, method for degrading vomitoxin and kit Download PDFInfo
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Abstract
The invention relates to the field of microorganisms, and discloses bacillus subtilis, a microbial inoculum containing the bacillus subtilis, application of the bacillus subtilis, a method for degrading vomitoxin and a kit. Specifically, the invention provides a Bacillus subtilis strain, and the preservation number of the Bacillus subtilis strain is CGMCC NO. 13139. The bacillus subtilis provided by the invention can efficiently and quickly degrade vomitoxin in grain oil and/or feed, and particularly, even if the content of the vomitoxin in the grain oil and/or feed is more than 50ppm, the bacillus subtilis can efficiently and quickly degrade the vomitoxin and has no influence on the palatability of the grain oil and/or feed; meanwhile, the bacillus subtilis is added into grain oil and/or feed polluted by vomitoxin, so that the bacillus subtilis has safety. Therefore, the bacillus subtilis has a good application prospect.
Description
Technical Field
The invention relates to the field of microorganisms, in particular to bacillus subtilis, a microbial inoculum containing the bacillus subtilis, application of the bacillus subtilis, a method for degrading vomitoxin and a kit.
Background
Vomitoxin (vomitoxin), also known as Deoxynivalenol (DON), is named after 3 α,7 α, 15-trihydroxy Fusarium solani-9-en-8-one, because it can cause vomiting of pigs, is mainly trichothecene toxins produced by Fusarium graminearum (Fusarium graminearum) and Fusarium yellow (Fusarium culmorum) infecting grains such as wheat, barley, oats and corn. Vomitoxin is one of main mycotoxin pollution of grains, feeds and foods worldwide, and seriously affects the health of people and livestock. After people and livestock ingest food polluted by vomitoxin, acute poisoning symptoms such as anorexia, vomit, diarrhea, fever, unstable standing, slow response and the like can be caused, the hematopoietic system is damaged in serious cases to cause death, and the serious harmfulness of the food has attracted general attention of various countries.
The vomitoxin has quite common pollution to grain raw materials in China, the detection rate and the detection amount of the vomitoxin are the highest one of mycotoxins, and investigation by screening sunshine and the like shows that the standard exceeding proportion of the vomitoxin of Chinese feed and raw materials is close to 70%, the standard exceeding rate of the vomitoxin in corn is 57.1%, the average content of the toxin is 1.01mg/kg, and the highest content of the vomitoxin is 2.13 mg/kg. Because of the ubiquitous nature, high content and acute and chronic toxicity of vomitoxin in grains and feeds, it is important and urgent to reduce or eliminate the toxicity. At present, the detoxification method of vomitoxin at home and abroad mainly comprises three major methods, namely a physical method, a chemical treatment and a biological method. Although detoxification by physical and chemical methods has been successful to a certain extent, the disadvantages of limited detoxification effect, possible loss of important nutrients, high cost and the like still exist, so that the application of the two methods in actual production is limited.
The biological method mainly utilizes microorganisms or degradation products thereof to carry out toxin degradation, has the advantages of reducing toxicity of toxins under mild conditions, having small influence on sensory properties, palatability and nutrient substances of raw materials and the like, and has the characteristics of safety, environmental protection and high efficiency, thereby being considered as the optimal detoxification method. Therefore, the research of removing vomitoxin in grain and oil or/feed by using modern biotechnology has good application prospect. The patent application CN103243047A discloses a bacillus subtilis for efficiently degrading vomitoxin and an application thereof, wherein 900 mu L of bacillus subtilis ANSB471 fermentation liquor reacts with 100 mu L of vomitoxin (100 mu g/ml), the degradation rate of the vomitoxin is 25% after 2 hours of reaction, the degradation rate of the vomitoxin is 56% after 24 hours of reaction, the degradation rate of the vomitoxin is 80% after 48 hours of reaction, and the degradation rate is yet to be improved.
In addition, the existing biological methods for degrading vomitoxin are mostly carried out under mild conditions (such as temperature 25-37 ℃ and not more than 40 ℃ at the maximum, and pH value is about 7), however, no good solution exists under higher temperature load (such as the condition of transportation in a container or during feed granulation) or under severe acid-base conditions, which limits the application range of the biological methods in degrading the vomitoxin.
Therefore, there is a need to find a method for biodegrading vomitoxin that is efficient and safe and can be used under high temperature load or harsh acid-base conditions.
Disclosure of Invention
The invention aims to overcome the defects in the existing vomitoxin degradation process, and provides a bacillus subtilis strain, a microbial inoculum containing the bacillus subtilis strain, application of the bacillus subtilis strain, a method for degrading vomitoxin and a kit.
In order to achieve the purpose, in a first aspect, the invention provides a Bacillus subtilis strain, wherein the preservation number of the Bacillus subtilis strain is CGMCC NO. 13139.
In a second aspect, the invention also provides a microbial inoculum, wherein the microbial inoculum contains the Bacillus subtilis.
In a third aspect, the invention also provides application of the bacillus subtilis and/or the microbial inoculum in degrading vomitoxin.
In a fourth aspect, the present invention also provides a method for degrading emetic toxin, wherein the method comprises: and (3) contacting the bacillus subtilis and/or the microbial inoculum with a vomitoxin-polluted sample to degrade the vomitoxin in the sample.
In a fifth aspect, the present invention provides a kit comprising the bacillus subtilis and/or the microbial inoculum of the present invention.
The bacillus subtilis provided by the invention can efficiently and quickly degrade vomitoxin in grain oil and/or feed, and particularly, even if the content of the vomitoxin in the grain oil and/or feed is more than 50ppm (preferably at least 100ppm, more preferably at least 150ppm, and even more preferably at least 200ppm), the bacillus subtilis can efficiently and quickly degrade the vomitoxin and has no influence on the palatability of the grain oil and/or feed (namely, the taste is basically not different before and after treatment); meanwhile, the bacillus subtilis is added into grain oil and/or feed polluted by vomitoxin, so that the bacillus subtilis has safety. Therefore, the bacillus subtilis has a good application prospect.
In addition, the bacillus subtilis provided by the invention also has high temperature and acid-base stability, so that the application range of the bacillus subtilis is further expanded.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Biological preservation
The Bacillus subtilis is preserved in the common microorganism center of China Committee for culture Collection of microorganisms (the address: No. 3 of the institute of microbiology, national academy of sciences, No.1 of West Lu, North Chen, south China, Beijing, the city of Tokyo, the abbreviation of the preservation unit is CGMCC), and the preservation number is CGMCC NO.13139, 24 days 10 and 24 days 2016.
Detailed Description
The following describes in detail specific embodiments of the present invention. It should be understood that the detailed description and specific examples, while indicating the present invention, are given by way of illustration and explanation only, not limitation.
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
In a first aspect, the invention provides a Bacillus subtilis strain, wherein the preservation number of the Bacillus subtilis strain is CGMCC NO. 13139.
The bacillus subtilis provided by the invention is separated from a soil sample (the collection place is Beijing) polluted by vomitoxin. The isolation of Bacillus subtilis may be carried out by a method conventionally used in the art for isolation of a novel strain, and may be, for example, a liquid phase enrichment method or a soil circulation method.
The liquid phase enrichment method may specifically include: weighing a proper amount of soil sample polluted by vomitoxin, adding the soil sample into a triangular flask filled with LB liquid culture medium, and adding a proper amount of glass beads. Oscillating the triangular flask at 28-37 ℃ and 160-; transferring the soil mixed solution into a centrifuge tube, and taking the centrifuged supernatant as a source of vomitoxin passivation microorganisms. The supernatant was inoculated into LB liquid medium containing vomitoxin and cultured with shaking at 28-37 ℃ and 160-180 rpm. Pipette 1-2mL with a sterile pipette and transfer to another enrichment culture flask. After three such transfers, the supernatants were diluted 10 times each-1、10-2、10-3、10-4、10-5、10-6、10-7Or 10-8Double, absorb and fitThe amount of the suspension was applied to LB solid medium containing 50, 100, 200 or 300mg/L of vomitoxin, streaked, and cultured at 28-37 ℃ for 3-4 days, followed by isolation of single colonies.
The soil circulation method may specifically include: 100g of soil polluted by vomitoxin is weighed and evenly mixed with a proper amount of sand grains with the grain diameter of about 3mm, and the mixture is placed on the upper layer of the circulation device. The lower layer was filled with 200mL of LB medium as a circulating solution. Starting an air compressor, starting the acclimatization process, and periodically supplementing the circulating fluid according to the evaporation condition of the circulating fluid. After the acclimatization is finished, respectively diluting the lower layer circulating fluid by 10-1、10-2、10-3、10-4、10-5、10-6、10-7Or 10-8Taking out appropriate amount of the suspension, spreading the suspension on LB solid medium containing 50, 100, 200 or 300mg/L vomitoxin, streaking, culturing at 28-37 deg.C for 3-4 days, and separating single colony.
The inventor selects a bacterium with the strongest degradation effect on vomitoxin from the screened strains to carry out DNA extraction and identification, and the identification result shows that the 16srDNA sequence of the strain has 100 percent homology with Bacillus subtilis, the strain can be determined to be Bacillus subtilis and preserved in the China general microbiological culture Collection center in 2016, 10, 24 and the preservation number is CGMCC NO. 13139.
The bacillus subtilis provided by the invention can produce a large amount of live bacteria of the bacillus subtilis after being cultured. The method of culturing the Bacillus subtilis of the present invention is not particularly limited as long as it can proliferate the Bacillus subtilis in a large amount by the culturing method, and for example, it can be 105Inoculating the live bacteria of the bacillus subtilis into a culture medium in the inoculation amount of CFU/mL, and culturing at the temperature of 28-38 ℃ for 12-48 hours under aerobic conditions to obtain a culture solution. Wherein the medium may be a medium conventionally used in the art, and for example, may be LB liquid medium (0.8-1 wt% peptone, 0.5-0.8 wt% yeast powder, 1-1.5 wt% sodium chloride; pH 6.8-7.0; culture temperature 28-38 ℃) or nutrient broth medium (0.8-1 wt% peptone, 0)3-0.5 wt% beef extract, 0.5-0.8 wt% sodium chloride; the pH value is 7.2-7.6; the culture temperature is 28-38 ℃), preferably LB liquid culture medium.
The present invention can further separate the bacillus subtilis cells from the culture solution, and the method for separating is not particularly limited as long as it can enrich the cells from the culture solution, and for example, the separation can be achieved by centrifugation and/or filtration, and the conditions for centrifugation and filtration can be conventional conditions in the art, which are well known to those skilled in the art and will not be described herein again.
In a second aspect, the present invention provides a bacterial agent, wherein the bacterial agent contains the Bacillus subtilis.
In the present invention, the concentration of bacillus subtilis in the microbial agent is not particularly limited, and may be specifically selected according to specific conditions.
According to the present invention, the microbial agent contains live cells and/or dead cells of the Bacillus subtilis. Preferably, the microbial inoculum contains live bacteria or mixed bacteria of live bacteria and dead bacteria of the bacillus subtilis. When the microbial agent contains a mixed body of live cells and dead cells of the Bacillus subtilis, the number of live cells is preferably higher than the number of dead cells. Most preferably, the microbial inoculum contains viable cells of the Bacillus subtilis.
According to the present invention, the formulation of the microbial inoculum is not particularly limited, and the microbial inoculum can be prepared into different formulations according to different intended uses, and can be added with corresponding components such as excipients, for example, the microbial inoculum can be a liquid microbial inoculum (for example, a bacterial solution) and/or a solid microbial inoculum (for example, a lyophilized microbial inoculum), preferably a solid microbial inoculum. The addition of excipients to the bacterial preparation in any dosage form is well known to those skilled in the art and will not be described in detail herein.
In addition, in the process of research, the inventor of the invention finds that although the bacillus subtilis provided by the invention is screened under the condition of high concentration of vomitoxin (at least 50ppm), the bacillus subtilis has a certain degradation effect on other mycotoxin pollutants (such as vomitoxin, aflatoxin, fumonisin, ochratoxin, T2 toxin and the like) commonly seen in soil.
In a third aspect, the invention also provides an application of the bacillus subtilis and/or the microbial inoculum in degrading vomitoxin, preferably an application in degrading vomitoxin in grain, oil and/or feed.
In a fourth aspect, the present invention also provides a method for degrading emetic toxin, wherein the method comprises: and (3) contacting the bacillus subtilis and/or the microbial inoculum with a vomitoxin-polluted sample to degrade the vomitoxin in the sample.
According to the invention, the vomitoxin-contaminated sample may be vomitoxin-contaminated grain oil and/or feed.
Preferably, the vomitoxin is present in an amount of at least 1ppm, more preferably at least 50ppm, even more preferably at least 100ppm, even more preferably at least 150ppm, even more preferably at least 200ppm, based on the total weight of the vomitoxin-contaminated sample. In the present invention, "ppm" means "μ g/mL" when the sample to be treated is a liquid; when the sample to be treated is a solid, "ppm" means "μ g/g".
In the present invention, the term "grain and oil" refers to a general term for grains, beans and other grains and oils, and finished products and semi-finished products thereof, and particularly to products that can be eaten by humans. For example, the grain oil may be a grain oil product that is edible to humans and is common in the art, and specifically, the grain oil may include at least one of grains and agricultural byproducts thereof, oil and fat products, wines, milks and products thereof, and the like.
In the present invention, the term "feed" refers to the general term of food for animals raised in agriculture or animal husbandry. For example, the feed may be a food commonly used in the art for feeding animals, and in particular, the feed may include: a) cereals, for example, small grain cereals (such as wheat, barley, rye, oats, and combinations thereof) and/or large grain cereals such as maize or sorghum; b) by-products from cereals, such as corn gluten meal, distillers dried grains with solubles (DDGS), wheat bran, wheat middlings, rice bran, rice hulls, oat hulls, palm kernel, and citrus pulp; c) ensiling the feed; d) proteins from the following sources: such as soy, sunflower, peanut, lupin, pea, broad bean, cotton, canola, fish meal, dried plasma protein, meat and bone meal, potato protein, whey, copra, sesame; e) oils and fats obtained from plant and animal sources; f) minerals and vitamins.
In the present invention, the grain or feed may further comprise a physiologically acceptable carrier, wherein the physiologically acceptable carrier is at least one selected from the group consisting of: maltodextrin, limestone (calcium carbonate), cyclodextrin, wheat bran or wheat component, rice or rice bran, sucrose, starch, Na2SO4And talc and mixtures thereof.
In the present invention, the formulation of the bacillus subtilis and/or the microbial inoculum is not particularly limited, and it can be prepared into different formulations according to the intended use, and corresponding excipients and other ingredients are added, for example, the bacillus subtilis and/or the microbial inoculum can be in liquid and/or solid state. Wherein, the addition of excipients to the dosage forms is well known to those skilled in the art, and will not be described in detail herein.
According to the present invention, the number of cells in the Bacillus subtilis and/or the microbial inoculum is at least 10 per 1g of the sample5CFU, preferably, the number of viable bacteria in the Bacillus subtilis and/or the microbial inoculum is at least 105And (4) CFU. In the present invention, the cell count can be measured according to GB 4789.2-94.
According to the present invention, the conditions of the contacting may include: the temperature is 25-55 ℃, the pH value is 3-9, and the time is 1-48 h; preferably, the temperature is 30-40 ℃, the pH value is 6-8, and the time is 12-36 h.
In the present invention, when the sample is a solid, the pH value is determined according to the method of GB/T12456-.
In a fifth aspect, the present invention provides a kit comprising the bacillus subtilis and/or the microbial inoculum of the present invention.
Examples
The present invention will be described in detail below by way of examples.
In the following preparations, preparation comparative examples, examples and comparative examples:
vomitoxin standards were purchased from Sigma, and the remaining experimental materials, unless otherwise specified, were purchased from conventional biochemical stores.
LB liquid medium: 10g of peptone, 5g of yeast powder, 10g of sodium chloride and deionized water are added to the solution to reach a constant volume of 1L, and the pH value is adjusted to 7.
LB solid medium: 10g of peptone, 5g of yeast powder, 10g of sodium chloride, 16g of agar powder and deionized water, wherein the volume is constant to 1L, and the pH value is adjusted to 7.
The strain provided by the invention is Bacillus subtilis, and is preserved in the common microorganism center of China general microbiological culture Collection center (address: No. 3 of West Lu No.1 of North Chen of the Korean-Yang district, Beijing, institute of microbiology, China academy of sciences, postal code: 100101) 24 days 10.24 days 2016 (the abbreviation of the preservation unit is CGMCC), and the preservation number is CGMCC NO. 13139.
The reference strain 1 is Bacillus subtilis (CGMCC NO. 7344) published in CN103243047A and purchased from CGMCC.
The reference strain 2 is Bacillus licheniformis (Bacillus licheniformis) with the preservation number of CGMCC NO.1.807, purchased from CGMCC.
The cell count was measured according to GB 4789.2-94.
Determining the content of vomitoxin according to the determination immunoaffinity column purification-high performance liquid chromatography of deoxynivalenol in GB/T30956-2014 feed.
The degradation rate (%) of vomitoxin (mass of vomitoxin in the sample before reaction-mass of vomitoxin in the sample after reaction)/mass of vomitoxin in the sample before reaction × 100%.
Preparation example 1
The bacillus subtilis with the preservation number of CGMCC NO.13139 provided by the invention is activatedThen, the cells were inoculated in an LB liquid medium sterilized at 121 ℃ for 15min at an inoculum size of 1 vol%, and cultured with shaking at 160rpm and 37 ℃ for a period of time such that the cell concentration in the cells was (1. + -. 0.1). times.105CFU/mL. Simultaneously preparing into dry powder, wherein the total viable count contained in each gram of dry powder is (1 + -0.1) x 105CFU。
Preparation of comparative example 1
According to the method of preparation example 1, except that Bacillus subtilis (Bacillus subtilis) having a preservation number of CGMCC NO.7344 disclosed in CN103243047A was used in place of the Bacillus subtilis having a preservation number of CGMCC NO.13139 provided by the present invention.
Preparation of comparative example 2
According to the method of preparation example 1, except that Bacillus licheniformis (Bacillus licheniformis) having a preservation number of CGMCC NO.1.807 was used in place of the Bacillus subtilis having a preservation number of CGMCC NO.13139 as provided in the present invention.
Example 1
This example illustrates the ability of bacillus subtilis to degrade emetic toxin provided by the present invention.
Firstly, measuring degradation time
Putting 900 mu L of the bacterial liquid cultured in the preparation example 1 into a 1.5mL centrifuge tube, adding the vomitoxin standard solution, and uniformly mixing to obtain a mixed solution, wherein the final concentration of the vomitoxin in the mixed solution is 50 ppm.
The mixture was reacted at 37 ℃ and pH 7, and 20. mu.L of the reaction sample was taken for 1 hour, 12 hours, 24 hours, 36 hours, and 48 hours, respectively, to detect the presence of vomitoxin. The results are shown in Table 1.
The results in Table 1 show that the reaction time of 24 hours can achieve a vomitoxin degradation rate of 90% or more. Therefore, 24h was used as the reaction time in the following experiment.
Measurement of thermal stability
Treating the cultured bacterial liquid at 40 deg.C, 45 deg.C, 50 deg.C and 55 deg.C, pH 7 for 30min, respectively placing 900 μ L of the treated bacterial liquid in 1.5mL centrifuge tube, adding vomitoxin standard solution, and mixing to obtain mixed liquid with final concentration of 50ppm of vomitoxin. Then, the mixture was reacted at 37 ℃ and pH 7 for 24 hours, and after completion of the reaction, 20. mu.L of the sample was used for detecting the vomitoxin residue. The results are shown in Table 2.
As can be seen from the results in Table 2, the bacterial solution still has high emetic toxin degradation activity when stored at 40-55 ℃ for 30 min.
③ determination of acid-base stability
Treating the cultured bacterial liquid at pH of 2, 3, 4, 5, 6, 7, 8 and 9 respectively at 37 deg.C for 30min, placing 900 μ L of the treated bacterial liquid in a 1.5mL centrifuge tube, adding vomitoxin standard solution, and mixing to obtain a mixed solution, wherein the final concentration of vomitoxin in the mixed solution is 50 ppm. Then, the mixture was reacted at 37 ℃ and pH 7 for 24 hours, and after completion of the reaction, 20. mu.L of the sample was used for detecting the vomitoxin residue. The results are shown in Table 3.
The results in Table 3 show that the bacterial liquid has high emetic toxin degradation activity even when stored for 30min at a pH of 3-9.
Comparative example 1
The measurement was carried out in the same manner as in example 1 except that the bacterial suspension prepared in preparation example 1 was used instead of the bacterial suspension used in example 1.
The results of the degradation time measurement are shown in table 1, the results of the thermal stability measurement are shown in table 2, and the results of the acid-base stability are shown in table 3.
Comparative example 2
The measurement was carried out in the same manner as in example 1 except that the bacterial suspension prepared in preparation example 2 was used instead of the bacterial suspension used in example 1.
The results of the degradation time measurement are shown in table 1, the results of the thermal stability measurement are shown in table 2, and the results of the acid-base stability are shown in table 3.
TABLE 1
TABLE 2
TABLE 3
Test example 1
10g of the dry powder obtained in preparation example 1 was mixed with 1kg of corn flour contaminated with 200ppm of vomitoxin, 1kg of distilled water was added, and three repetition was performed, and after uniform mixing, detoxification was performed for 1 day at 37 ℃ and pH 7. And then determining the content of vomitoxin in each sample after 1 day of treatment according to the determination immunoaffinity column purification-high performance liquid chromatography of deoxynivalenol in GB/T30956-2014 feed, and calculating the degradation rate. The degradation rate of vomitoxin in the corn flour is calculated to be 90.2%. Also, the addition of dry powder did not have any effect on the palatability of the corn meal.
The results of feeding pigs with the treated corn meal for 14 days show that the diet, drinking water and daily activities of the pigs are normal during the period, and the weight of the pigs always keeps on a stable increasing trend. The above results show that the dry powder obtained in preparation example 1 is added to corn flour for pig feeding, and is safe.
Test example 2
According to the method of test example 1, except that the content of vomitoxin in the corn flour was 100ppm, the degradation rate of vomitoxin was detected to be 95.2%. Also, the addition of dry powder did not have any effect on the palatability of the corn meal.
The results of feeding pigs with the treated corn meal for 14 days show that the diet, drinking water and daily activities of the pigs are normal during the period, and the weight of the pigs always keeps on a stable increasing trend. The above results demonstrate the safety of adding dry powder to corn meal for pig feeding.
Test example 3
According to the method of test example 1, except that the content of vomitoxin in the corn flour was 50ppm, the degradation rate of vomitoxin was 100%. Also, the addition of dry powder did not have any effect on the palatability of the corn meal.
The results of feeding pigs with the treated corn meal for 14 days show that the diet, drinking water and daily activities of the pigs are normal during the period, and the weight of the pigs always keeps on a stable increasing trend. The above results demonstrate the safety of adding dry powder to corn meal for pig feeding.
Test comparative example 1
The procedure of test example 1 was followed, except that the dry powder obtained in preparation comparative example 1 was used in place of the dry powder used in example 1. The degradation rate of the vomitoxin is 18.1 percent through detection.
Test comparative example 2
The procedure of test example 1 was followed except that the dry powder obtained in preparation comparative example 2 was used in place of the dry powder used in example 1. The detected degradation rate of the vomitoxin is 16.8%.
Compared with the results of the comparative examples 1-2, the bacillus subtilis provided by the invention can effectively and quickly degrade vomitoxin in grain, oil and/or feed, and has high temperature and acid-base stability.
As can be seen from the comparison of the results of the above test example 1 and the test comparative examples 1-2, even when the bacillus subtilis provided by the present invention is applied to grain oil and/or feed with vomitoxin content as high as 50ppm or more, the bacillus subtilis can degrade the vomitoxin efficiently and rapidly, and has no influence on the palatability of the grain oil and/or feed (i.e., there is substantially no difference in taste before and after treatment); meanwhile, the bacillus subtilis is added into grain oil and/or feed polluted by vomitoxin, so that the bacillus subtilis has safety. Therefore, the bacillus subtilis has a good application prospect.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various features described in the above embodiments may be combined in any suitable manner without departing from the scope of the invention. The invention is not described in detail in order to avoid unnecessary repetition.
In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.
Claims (13)
1. Bacillus subtilis (B.subtilis)Bacillus subtilis) The bacillus subtilis is characterized in that the preservation number of the bacillus subtilis is CGMCC number 13139.
2. A bacterial agent comprising the Bacillus subtilis (Bacillus subtilis) of claim 1Bacillus subtilis）。
3. The microbial agent according to claim 2, wherein the microbial agent contains a living cell of the Bacillus subtilis.
4. The microbial inoculum according to claim 2 or 3, wherein the microbial inoculum is a liquid microbial inoculum or a solid microbial inoculum.
5. The microbial inoculum according to claim 4, wherein the microbial inoculum is a solid microbial inoculum.
6. Use of the bacillus subtilis of claim 1 and/or the bacterial agent of any one of claims 2 to 5 for degrading emetic toxin.
7. The use of claim 6, wherein the Bacillus subtilis and/or the microbial inoculum is used for degrading vomitoxin in grain oil and/or feed.
8. A method of degrading emetic toxin, the method comprising: contacting the bacillus subtilis of claim 1 and/or the bacterial agent of any one of claims 2 to 5 with a vomitoxin-contaminated sample to degrade the vomitoxin in the sample.
9. The method of claim 8, wherein the vomitoxin-contaminated sample is vomitoxin-contaminated grain oil and/or feed.
10. The method according to claim 8 or 9, wherein the number of cells in the Bacillus subtilis and/or the microbial agent is at least 10 relative to 1g of the sample5CFU。
11. The method of claim 8 or 9, wherein the conditions of the contacting comprise: the temperature is 25-55 deg.C, and pH is 3-9.
12. The method of claim 11, wherein the conditions of the contacting comprise: the temperature is 30-40 deg.C, and pH is 6-8.
13. A kit comprising the Bacillus subtilis of claim 1 and/or the microbial agent of any one of claims 2 to 5.
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| CN111328962B (en) * | 2018-12-19 | 2022-10-21 | 吉林中粮生化有限公司 | Method for degrading vomitoxin in corn deep processing process |
| CN109913388B (en) * | 2019-03-20 | 2019-12-31 | 中粮集团有限公司 | Composite microbial inoculum for improving corn soaking effect and application thereof |
| CN109744367A (en) * | 2019-03-22 | 2019-05-14 | 东莞市澹一生物科技有限公司 | The method and feedstuff of resource utilization wheat vinasse production feedstuff |
| CN109744368A (en) * | 2019-03-22 | 2019-05-14 | 东莞市澹一生物科技有限公司 | Utilize the method and feedstuff of wheat vinasse production feedstuff |
| CN111304119B (en) * | 2020-02-21 | 2021-12-07 | 江苏澳华生物科技研究院有限公司 | Feeding bacillus subtilis for degrading fumonisins and application thereof |
| CN111763641A (en) * | 2020-07-01 | 2020-10-13 | 中国农业科学院植物保护研究所 | Bacillus subtilis capable of efficiently degrading vomitoxin and application thereof |
| CN112458026A (en) * | 2020-12-15 | 2021-03-09 | 江苏奥迈生物科技有限公司 | Bacillus subtilis capable of efficiently degrading vomitoxin and application thereof |
| CN113980838B (en) * | 2021-09-30 | 2023-07-28 | 安阳中农大生物技术有限公司 | Bacillus subtilis for efficiently and directionally expressing bacteriocin M6 and application thereof |
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