Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Bacillus subtilis C3 and its anti-Listeria monocytogenes bacteriocin preparation method

A technology of Bacillus subtilis and Listeria monocytogenes, which is applied in the field of microbiology, can solve problems such as no related reports, and achieve the effect of simple extraction process, wide sources and efficient purification method

Active Publication Date: 2013-09-25
BEIJING UNIV OF AGRI
View PDF6 Cites 12 Cited by
  • 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 Bacillus subtilis bacteriocin against Listeria monocytogenes in domestic invention patents and literature

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Bacillus subtilis C3 and its anti-Listeria monocytogenes bacteriocin preparation method
  • Bacillus subtilis C3 and its anti-Listeria monocytogenes bacteriocin preparation method
  • Bacillus subtilis C3 and its anti-Listeria monocytogenes bacteriocin preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, the preparation of bacillus subtilis C3 bacteriocin crude extract

[0026] 1. Preparation of Bacillus subtilis C3 fermentation broth

[0027] (1) Preparation of seed solution

[0028] Seed medium formula (W / V, mass volume ratio concentration): glucose 1%, peptone 1%, NaCl 1%, KH 2 PO 4 0.15%, MgS0 4 ·7H 2 O0.15%, natural pH;

[0029] Seed culture conditions: transfer the activated Bacillus subtilis into the seed medium according to the inoculum amount of 1%, the culture temperature is 37°C, the liquid volume is 75mL / 250mL, shake the shaker at 150r / min for 18h, and the number of viable bacteria is about 10 9 CFU / mL.

[0030] (2) Preparation of fermentation broth

[0031] Fermentation medium formula (W / V, mass volume ratio concentration): glucose 1%, peptone 1.5%, yeast extract powder 1.5%, KH 2 PO 4 0.1%, MgSO 4 ·7H 2 O0.15%, NaCl0.5%, pH7.0;

[0032] Fermentation conditions: transfer the seed liquid into the fermentation medium according to the...

Embodiment 2

[0036] Example 2, Bacillus subtilis C3 bacteriocin crude extract antibacterial properties and physicochemical properties

[0037] During the test, the antibacterial properties and physicochemical properties of the crude extract of C3 bacteriocin were determined by the tube-and-disk method.

[0038] Tube-and-dish method: first dilute the indicator strain to 10 7 CFU / mL, mix with the corresponding solid medium that has been heated and melted, pour about 15mL into a plate, place a sterile Oxford cup gently after solidification, take 100μL of Bacillus subtilis C3 crude extract into the Oxford cup, and incubate at 37°C After 12 hours, observe the inhibition zone, and measure the diameter of the inhibition zone with a vernier caliper, and the reading is accurate to 0.01mm.

[0039] 1. The antibacterial properties of the crude extract of Bacillus subtilis C3 bacteriocin

[0040] The antibacterial effect of table 1 Bacillus subtilis C3 bacteriocin crude extract

[0041]

[0042]...

Embodiment 3

[0062] Embodiment 3, Bacillus subtilis C3 bacteriocin purification method

[0063] 1. Purification of Bacillus subtilis C3 bacteriocin crude extract

[0064] (1) Step-by-step elution and purification by anion exchange chromatography

[0065] The crude extract of Bacillus subtilis C3 bacteriocin was filtered and sterilized with a 0.2 μm sterile filter, and then eluted and purified step by step by DEAE-Sepharose Fast Flow anion exchange chromatography, with a chromatographic column size of 10mm×200mm; buffer: A Solution 0.025mol / L pH7.5Tric-HCl, B solution contains 0.025mol / L pH7.5Tric-HCl with 0.4mol / L NaCl; stepwise elution step: 50min100%A solution→90min70%A solution+30%B solution → 40min 50% solution A + 50% solution B → 40min 100% solution B; the flow rate is 1mL / min, the sample volume is 5mL, the automatic collector is 5mL / tube, and the collected eluent is detected by the tube-and-disk method against Listeria monocytogenes Antibacterial activity, the test showed that the...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Diameteraaaaaaaaaa
Diameteraaaaaaaaaa
Diameteraaaaaaaaaa
Login to View More

Abstract

The name of the invention is a bacillus subtilis C3 and its anti-Listeria monocytogenes bacteriocin preparation method. The bacteriocin, used as a natural antiseptic, is suitable for low temperature meat and meat product and fruit and vegetable fresh-keeping, and can be used to raise safety of food. According to the invention, Bacillus subtilis C3 CGMCC No.9713 is utilized by ammonium sulfate precipitation, anion exchange chromatography and semi-preparative reversed phase liquid chromatography for fine purification. Thus, titer of the bacillus subtilis C3 bacteriocin is raised by 32 times, and purity is raised by 42.80 times. The bacteriocin has a very strong bacteriostatic activity of Listeria monocytogenes and good heat and pH stability, can be degraded by protease K, and is a natural safe biological preservative. The bacteriocin provided by the invention has advantages of wide sources of raw materials for production, simple extraction technology and efficient and stable purification method. The preparation method provided by the invention is suitable for industrial production.

Description

technical field [0001] The invention relates to a preparation method of Bacillus subtilis C3 and anti-Listeria monocytogenes bacteriocin in the field of microorganisms. As a natural preservative, the bacteriocin is suitable for keeping low-temperature meat and meat products, fruits and vegetables fresh, and can improve food safety. Background technique [0002] During the process of processing, storage, transportation, etc., food is easily contaminated by harmful microorganisms, resulting in spoilage or food poisoning. The method of adding preservatives to inhibit the growth of microorganisms has gradually become an important means of food preservation. Traditional meat products are mostly preserved by chemical preservatives, which have certain safety hazards. Improper use will have certain side effects. Long-term excessive intake will cause certain damage to consumers’ bodies and lead to environmental pollution. Therefore, the development of efficient, safe and environment...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N1/20C07K14/32C07K1/36C07K1/30C07K1/20C07K1/18C12R1/125
Inventor 谢远红刘慧张红星杨永青熊利霞高秀芝马晓丹
Owner BEIJING UNIV OF AGRI
Features
  • Generate Ideas
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com