A Strain of Lactococcus lactis CAMT22361 Degrading t-2 Toxin

A technology of CAMT22361 and Lactococcus lactis, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., can solve the problems of secondary pollution, chemical reagent residues, etc., and achieve good application prospects

Active Publication Date: 2019-08-09
山东宝来利来生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally speaking, this type of method is better than the physical method, but there may be some residual chemical reagents, causing secondary pollution

Method used

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  • A Strain of Lactococcus lactis CAMT22361 Degrading t-2 Toxin
  • A Strain of Lactococcus lactis CAMT22361 Degrading t-2 Toxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Isolation and identification of strains

[0018] Isolation of lactic acid bacteria: Rinse the surface of sandworms with sterilized seawater, dry them, weigh 25g of intestinal samples and cut them into pieces. Diluted by 10-fold dilution method, and spread on MRS solid medium containing 1.5% calcium, observed after culturing at 25 °C for 3-5 days, picked the colonies with calcification circles, purified, and then carried out Gram staining. and catalase test. Gram-positive and catalase-negative strains were identified as lactic acid bacteria, and slanted was stored.

[0019] Screening of lactic acid bacteria degrading T-2 toxin: inoculate the isolated lactic acid bacteria into the modified MRS liquid medium containing 50 ng T-2 toxin per ml, (modified MRS medium formula g / L: peptone 10 g, beef extract 10g, K 2 HPO 4 2g, Diammonium Citrate 2g, Sodium Acetate 5g, Glucose 20g, Tween-80l mL, MgSO 4 ·7H 2 O 0.58g, MnSO 4 ·4H 2 O0.25g, distilled water 1000mL,...

Embodiment 2

[0024] 1. Activation of Lactococcus lactis CAMT22361

[0025] In an ultra-clean operating table, the above-mentioned Lactococcus lactis CAMT22361 was picked and inoculated into the MRS liquid medium (pH 6.2-6.4) improved in Example 1, and cultivated in a shaking table at 37°C for 24 hours to obtain an activated bacterial liquid.

[0026] 2. Inoculation of fermentation broth

[0027] In an ultra-clean operating table, take a certain amount of activated bacterial solution, centrifuge at 10,000 r / min for 2 min, discard the supernatant, wash the pellet with sterile saline twice, and resuspend to the original volume. Use sterile water as a blank control to measure OD. 600 nm, and diluted to absorbance value of 0.3 with sterile physiological saline to prepare bacterial suspension, and inoculated the bacterial suspension on the modified MRS liquid medium at an inoculum of 1%, so that the initial bacteria in the broth medium were The number is about 10 8 CFU / mL.

[0028] 3. Addit...

Embodiment 3

[0047] Example 3: Degradation effect of Lactococcus lactis CAMT22361 on T-2 toxin in feed of Penaeus vannamei

[0048] 1. Preparation of Lactococcus lactis CAMT22361 bacterial suspension

[0049] In an ultra-clean operating table, pick Lactococcus lactis CAMT22361 stored on a slanted surface and inoculate it in MRS broth medium, and cultivate at 37°C for 24-28 hours, so that the final concentration of the bacterial suspension reaches 10 8 CFU / mL.

[0050] 2. Preparation of T-2 toxin venom

[0051] Accurately weigh 1 mg of T-2 toxin standard (Enzo, USA, purity ≥ 98%), dissolve it in acetonitrile, dilute to 1 mL, and prepare 1 mg / mL T-2 toxin venom.

[0052] 3. Preparation of poisonous bait for Penaeus vannamei

[0053] Prawn basic feed (33% imported fish meal, 4% squid meal, 18% soybean meal, 8% peanut meal, 4% shrimp bran, 1% mineral premix, 22% high-gluten flour, 1% fish oil, 3% soybean lecithin %, premix 5.7%, multivitamin 0.3%, crushed until all feeds pass through a 40-...

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Abstract

The invention belongs to the technical field of toxin biological treatment, and specifically discloses a strain of Lactococcus lactis capable of degrading T-2 toxin ( Lactococcus lactis ) CAMT22361, the Lactococcus lactis Lactococcus lactis CAMT22361 was deposited in China Center for Type Culture Collection (CCTCC) on May 13, 2015, and the deposit number is CCTCC NO: M 2015295. The Lactococcus lactis Lactococcus lactis The degradation rate of T-2 toxin can reach 61% after the fermentation broth of CAMT22361 reacts with T-2 toxin for 72 hours. When it is applied to remove T‑2 toxin in feed, the degradation rate can reach more than 50%. Lactococcus lactis of the present invention Lactococcus lactis CAMT22361 has the ability to efficiently and rapidly degrade T-2 toxin, and has a good application prospect in the treatment of T-2 toxin in biodegradable feed.

Description

technical field [0001] The invention belongs to the technical field of toxin biological treatment, and more particularly relates to a strain of Lactococcus lactis CAMT22361 that degrades T-2 toxin. Background technique [0002] T-2 toxin is 4,15-diacetoxy-8-(isovaleryloxy)-12,13-epoxy trichothecene-9-en-3 alcohol, which belongs to sesquiterpenes, It is a typical representative of trichothecene toxins. Toxic secondary metabolites mainly produced by fungi such as Fusarium sporotricoides, Fusarium equiseti, Fusariumpose, and Fusarium acuminatum. T-2 toxin can have toxic effects on various tissues and organs of humans and animals through different ways, especially to actively proliferating cells such as bone marrow, liver, lymphocytes and mucosal epithelial cells, which can cause teratogenicity and even carcinogenicity. In 1973, the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO) classified T-2 toxin as the most dangerous s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L5/20C12R1/46
CPCC12N1/205C12R2001/46
Inventor 刘颖徐春厚王雅玲孙力军
Owner 山东宝来利来生物工程股份有限公司
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