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Lactobacillus delbrueckiisubsp.lactis and preparation method of bacteriocin thereof capable of resisting Listeria monocytogenes

A technology of Lactobacillus delbrueckii and lactic acid subspecies, applied in the field of microorganisms, to achieve the effect of simple extraction process, simple operation and convenient source

Active Publication Date: 2012-10-24
BEIJING UNIV OF AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant report on the preparation method of Lactobacillus delbrueckii subspecies bacteriocin in domestic invention patents and literature

Method used

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  • Lactobacillus delbrueckiisubsp.lactis and preparation method of bacteriocin thereof capable of resisting Listeria monocytogenes
  • Lactobacillus delbrueckiisubsp.lactis and preparation method of bacteriocin thereof capable of resisting Listeria monocytogenes
  • Lactobacillus delbrueckiisubsp.lactis and preparation method of bacteriocin thereof capable of resisting Listeria monocytogenes

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Experimental program
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Embodiment 1

[0029] The percentages in the following examples are volume percentages unless otherwise specified. Example 1. Screening and identification of lactic acid bacteria strain zhang-LF with anti-Listeria monocytogenes activity

[0030] 1. Screening of lactic acid bacteria strain zhang-LF with anti-Listeria monocytogenes activity

[0031] (1) Preliminary screening of lactic acid bacteria strain zhang-LF with anti-Listeria monocytogenes activity

[0032] Take 1 g of the ground dried fish product powder and put it into 9 mL of sterile saline, shake it fully for 20 min to make a suspension, and dilute it 10 times to 10 -5 ~10 -8 Spread the sample suspensions of each dilution on the MRS selective medium containing calcium carbonate, incubate at 37°C for 24 hours, and then carry out Gram staining, depending on the individual shape and purity. Transfer to inoculate in MRS liquid medium, enrich the bacteria for 2 to 3 generations at 37°C, centrifuge to get the fermentation supernatant, ...

Embodiment 2

[0068] Embodiment 2, Lactobacillus delbrueckii lactic acid subspecies zhang-LF bacteriocin extraction method

[0069] 1. The fermentation process conditions of bacteriocin

[0070] Medium components (w / v) for culturing Lactobacillus delbrueckii lactic acid subsp. zhang-LF producing bacteriocin: tryptone 1%, beef extract 1%, yeast extract powder 0.5%, triammonium citrate 0.2%, glucose 2%, Tween-80 0.1%, sodium acetate 0.5%, dipotassium hydrogen phosphate 0.2%, magnesium sulfate 0.05%, manganese sulfate 0.025%, 0.07MPa sterilization for 20min.

[0071] Fermentation process conditions for producing bacteriocin: fermentation temperature 37° C., fermentation time 20 h, initial pH of the culture medium 6.0, inoculum size 1%.

[0072] 2. Extraction method and potency analysis of bacteriocin in fermentation broth

[0073] (1) Extraction method of bacteriocin The purpose of separation and purification of bacteriocin is to extract the target protein and purify it to reach the required...

Embodiment 3

[0082] Example 3, a simple and effective purification method of Lactobacillus delbrueckii lactic acid subspecies zhang-LF bacteriocin

[0083] 1. Purification method of Lactobacillus delbrueckii lactic acid subsp. zhang-LF bacteriocin

[0084]The crude bacteriocin was redissolved in citric acid-sodium citrate buffer at pH 3.0, and the bacteriocin was purified by SP SepharoseFast Flow cation exchange chromatography; the elution condition of cation exchange chromatography was: the flow rate was 1ml / min, pH 3.0 citric acid-sodium citrate buffer containing 0.1-0.5NaCl for linear gradient elution, collect the solution with a large absorption peak at 280nm; put the collected solution into a pretreated dialysis bag at 4°C Dialyze in deionized water for 24 hours to remove salt, and the dialysate is the purified product of bacteriocin.

[0085] 2. Evaluation of bacteriocin purification effect

[0086] The total protein content was determined by Coomassie Brilliant Blue G-250 staining...

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Abstract

The invention relates to a kind of Lactobacillus delbrueckiisubsp.lactis and a preparation method of a bacteriocin thereof capable of resisting Listeria monocytogenes, and the bacteriocin is suitable for the antisepsis and freshness of meat and meat products, milk and dairy products, fruits and vegetables, ready-to-use food, and the like. In the preparation method, the Lactobacillus delbrueckiisubsp.lactis zhang-LFCGMCC No.4393 is utilized and an adsorption-desorption method depending on the pH value and the cationic exchange chromatography are utilized to prepare the bacteriocin, thus the titer is increased by 32 times and the purity is increased by 25.23 times; and the bacteriocin has high antibacterial activity to Listeria monocytogenes and better thermal stability and acidic and alkaline stability and can be degraded by human protease, thus the bacteriocin is a natural and safe biopreservative. The invention provides a simple and effective preparation method of the Lactobacillus delbrueckiisubsp.lactis bacteriocin capable of resisting the activity of Listeria monocytogenes; and the extraction and purification methods are simple, the cost is lower, the demand on equipment is low and the method is suitable for industrial production.

Description

technical field [0001] The invention relates to a preparation method of Lactobacillus delbrueckii lactic acid subspecies and anti-Listeria monocytogenes bacteriocin in the field of microorganisms. The bacteriocin is used as a natural preservative and is suitable for meat and meat products, milk and dairy products, fruits and vegetables, Improving food safety in ready-to-eat foods. Background technique [0002] During the process of processing, storage, transportation, etc., food is easily contaminated by harmful microorganisms, resulting in spoilage or food poisoning. Adding preservatives to inhibit the growth of microorganisms is one of the important methods for food preservation today. Among the existing antiseptic methods, chemical preservatives are potentially harmful to the human body, while physical sterilization has a certain impact on the sensory and nutritional quality of food. As people pursue "more natural" and "minimally processed" foods, scholars have been stim...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P21/00C07K1/18C07K1/14C12R1/225
Inventor 张红星刘慧刘丽谢远红
Owner BEIJING UNIV OF AGRI