Pediococcus acidilactici capable of degrading fumonisins and application of pediococcus acidilactici

Abstract

The invention discloses pediococcus acidilactici capable of degrading fumonisins and application of the pediococcus acidilactici, the name of the pediococcus acidilactici is HAO2018, and the preservation number of the pediococcus acidilactici is CGMCC (China General Microbiological Culture Collection Center) 16658. The pediococcus acidilactici disclosed by the invention has a function of degrading fumonisins, is relatively high in degradation capacity, and can be independently used as a detoxification agent. The fumonisins degradation efficiency of the strain can reach 78.74%. In an M17 culture medium, the strain has a good growth rate. The optimal conditions of the strain for detoxification of fumonisins are as follows: the inoculum size is 2%, the initial pH is 6.5, the temperature is 37 DEG C, and the culture time is 36 hours. The pediococcus acidilactici disclosed by the invention has the capability of efficiently degrading fumonisins, and has a good application prospect in the industries of biodegradable feeds, food production and processing and the like.

Description

Technical Field

[0001] The invention relates to a Pediococcus acidilactici strain capable of degrading fumonisin and application of the strain in degrading fumonisin. Background Art

[0002] Fumonisins are mainly water-soluble secondary metabolites produced by Fusarium moniliforme, Fusarium verticillioides and Fusarium vesiculosus under certain conditions, mainly in the following categories: A, B, C, P, etc. Among them, fumonisins FB 1 The pollution range is the widest, the toxicity is the strongest, and the pollution level is the highest. Fumonisin can not only induce apoptosis and lesions in animals, but also induce liver cancer, esophageal cancer, lung cancer and other cancers in humans. Fumonisin has been listed as a Class 2B carcinogen by the International Agency for Research on Cancer. Therefore, effective prevention and control of fumonisin pollution is an urgent problem to be solved.

[0003] Studies have found that many microorganisms have a degrading effect on fumonisins. After the bacteria combine with fumonisins to form a complex, it is then excreted from the body, reducing the contact between the toxin and the gastrointestinal tract, reducing the absorption of the toxin, and protecting the body from harm. Compared with physical and chemical methods, microbial biological methods for degrading fumonisins have the advantages of being pollution-free and highly specific and have attracted much attention. However, microbial degradation of fumonisins still faces the problem of low efficiency and the inability of the strain to be directly applied.

[0004] The present invention discloses a strain of Pediococcus acidilactici that can effectively degrade fumonisin, which belongs to probiotics. After 2 hours of treatment, the 2018 strain has a fumonisin degradation capacity of 75.96%, and a 12-hour degradation efficiency of 78.74%, which has good application prospects in the application of degrading fumonisin. The use of probiotics such as lactic acid bacteria to adsorb fumonisin has good application prospects. In the food industry and feed processing industries, probiotics that can adsorb fumonisin are added. It has good application prospects in both the development of new biodegradable bacterial agents and biodegradable sterile preparations. Summary of the invention

[0005] The purpose of the present invention is to provide a strain of Pediococcus acidilactici with strong ability to degrade fumonisin, and application thereof in degrading fumonisin.

[0006] The lactic acid Pediococcus HAO2018 provided by the present invention is determined to belong to lactic acid Pediococcus through morphological and nucleic acid sequence analysis, and was deposited in the General Microbiological Center of China Microbiological Culture Collection Administration on October 30, 2018, with a deposit number of CGMCC No.16658. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Figure 1 This is a colony morphology diagram of Pediococcus acidilactici HAO2018 in the present invention;

[0008] Figure 2 This is the morphological diagram of Pediococcus acidilactici HAO2018 observed by Gram staining in the present invention;

[0009] Figure 3 This is the degradation rate of fumonisin by Pediococcus acidilactici HAO2018 at different times in the present invention. Detailed implementation manners

[0010] The following further illustrates the present invention in conjunction with implementation examples. The following are narrative implementations and are not restrictive. The protection scope of the present invention cannot be limited by the following examples.

[0011] The raw materials used in the present invention are all conventional commercially available products without special instructions. The methods used in the present invention are all conventional methods in the art without special instructions. The masses of various substances used in the present invention are all conventional masses for use.

[0012] A Pediococcus acidilactici capable of degrading fumonisin, characterized in that: the strain is named HAO2018, the classification name: Pediococcus acidilactici, the preservation number: CGMCC No. 16658, the preservation date: October 30, 2018, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, the preservation unit: China General Microbiological Culture Collection Center.

[0013] Preferably, the colonies of the Pediococcus acidilactici on the solid plate are white, round, with a smooth and moist surface, neat and raised edges, and reach the late logarithmic phase after being cultured in M17 medium for 24 hours under the condition of constant temperature at 37°C.

[0014] Example 1 Strain identification

[0015] (1) Materials and methods

[0016] M17 liquid medium (Qingdao Haibo Biotechnology Co., Ltd.), M17 solid medium (Qingdao Haibo Biotechnology Co., Ltd.), fumonisin standard FB 1 (Qingdao Prubio Biotechnology Co., Ltd.), phosphate buffered saline (PBS), and the medium was sterilized at 115°C for 20 minutes.

[0017] (2) Strain culture

[0018] The strain HAO2018 was inoculated into M17 solid medium and anaerobically cultured at 37°C for 24 hours. The colony morphology was observed to be white, round, with a smooth and moist surface, neat edges, and raised. The strain HAO2018 was inoculated into M17 liquid medium with an inoculation amount of 2%, and statically cultured for 36 hours.

[0019] (3) Strain identification

[0020] The obtained strain was subjected to PCR amplification of 16S rRNA, the PCR product was subjected to DNA sequence sequencing, and the obtained 16S rDNA sequence was subjected to BLAST homology alignment in the GenBank database to determine that the bacterial species was Pediococcus acidilactici.

[0021] Example 2 Study on the degradation ability of Pediococcus acidilactici HAO2018 to fumonisin

[0022] (1) Fumonisin standard stock solution: Weigh an appropriate amount of fumonisin standard FB 1 , dissolve it with 50% acetonitrile aqueous solution, and prepare a fumonisin FB 1 standard solution with a concentration of 50 μg / mL, and store it at -4°C;

[0023] (2) Sample preparation: Mix 45 μL of the bacterial liquid after fermentation of Pediococcus acidilactici HAO2018 with 5 μL of the fumonisin FB 1 standard stock solution, react in a water bath at 37°C in the dark for 20 min, and inactivate at 100°C for 10 minutes to terminate the reaction after the reaction;

[0024] (3) Sample derivatization: Add 200 μL of 50% acetonitrile water and 250 μL of OPA derivatization solution to the reaction solution to make the final concentration of fumonisin FB 1 10 μg / ml, mix well, filter through a 0.22 μm filter membrane, and perform HPLC detection within 2 minutes. By comparing with the peak map of the fumonisin FB 1 standard product, the degradation activity of Pediococcus acidilactici HAO2018 was determined.

[0025] Fumonisin degradation rate: It is expressed by the ratio of the reduced peak area m 1 of fumonisin FB 0 to the peak area m of the fumonisin FB 1 standard product;

[0026] Degradation situation: After 2 hours of treatment, the degradation ability of strain 2018 to fumonisin reached 75.96%. After 4, 6, and 12 hours of treatment, the degradation efficiency was relatively stable. The degradation efficiency at 4 hours was 76.21%, the degradation efficiency at 6 hours was 78.59%, and the degradation efficiency at 12 hours was 78.74%. Pediococcus lactis HAO2018 could stably and rapidly degrade fumonisin.

Claims

1. A Pediococcus acidilactici capable of degrading fumonisins, characterized in that: The strain was named HAO2018, the classification name: Pediococcus acidilactici, the preservation number: CGMCC No.16658, the preservation date: October 30, 2018, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, the preservation unit: General Microbiology Center of China Culture Collection Administration.

2. The bacterial agent, feed additive or animal feed of Pediococcus acidilactici HAO2018 according to claim 1.

3. The drug for Pediococcus acidilactici HAO2018 according to claim 1; the drug is a drug that inhibits fumonisin.

4. Use of Pediococcus acidilactici HAO2018 according to claim 1 in reducing fumonisin contamination in food or feed.

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