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Method for preparing bacteriocin and feruloyl esterase through solid-state fermentation of bacillus coagulans

A technology of Bacillus coagulans and ferulic acid esterase is applied in the field of solid-state fermentation of Bacillus coagulans to prepare bacteriocin and ferulic acid esterase, and can solve the problem of high cost of production raw materials, limited application of ferulic acid esterase, low yield, etc. question

Inactive Publication Date: 2021-06-08
XUZHOU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The problems of high cost of raw materials and low output faced by the ferulic acid esterase production industry have gradually become prominent, which greatly limits the application of ferulic acid esterase in various industries

Method used

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  • Method for preparing bacteriocin and feruloyl esterase through solid-state fermentation of bacillus coagulans
  • Method for preparing bacteriocin and feruloyl esterase through solid-state fermentation of bacillus coagulans
  • Method for preparing bacteriocin and feruloyl esterase through solid-state fermentation of bacillus coagulans

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A method for cultivating bacillus coagulans, comprising the steps of:

[0035] Step 1. Soil sample suspension

[0036] Take 50 soil samples from different garlic plantation soils in Quanshan District, Xuzhou City, Jiangsu Province, weigh 10g of each soil sample, place in a triangular flask equipped with glass beads and 100mL sterile water, and use a fermentation shaker at 120r / Rotate and oscillate for 1 hour to mix evenly, then place the flask in a shaking water bath at 80°C, and incubate with shaking for 35 minutes to obtain a soil sample suspension;

[0037] Step 2. Enrichment culture

[0038] Draw 2mL of the suspension from the Erlenmeyer flask of the soil sample suspension in Step 1 into a 250ml Erlenmeyer flask containing 30ml of enriched medium, culture it on a shaking table at 180r / min for 2 days at 35°C, and put the above culture solution at 5000r / min centrifuged for 15min, collected the bacteria under aseptic operation, then added sterile water to fully blo...

Embodiment 2

[0059] A method for preparing bacteriocin and ferulic acid esterase by solid-state fermentation of bacillus coagulans, the specific steps are as follows:

[0060] Step 1, preparation of liquid starter

[0061] a) Bacteria activation: Take the Bacillus coagulans strain preserved in glycerol and put it on the beef extract peptone medium for the first time, culture it at 35°C for 40 hours, pick a single colony from the above medium and repeat the culture on the new beef extract peptone again Streak again on the base, cultivate under the same conditions as above, and obtain a single colony for future use;

[0062] b) First-level seed liquid medium culture: pick a single colony from the medium for the second streak culture to the seed liquid medium, and the culture conditions are: 15mL seed liquid medium in a 150mL triangular bottle, pH 7, inoculate one Ring the bacteria, culture in a shaking incubator at 35°C at 180r / min for 18 hours, and obtain the first-grade seed liquid;

[0...

Embodiment 3

[0076] Separation and extraction of bacteriocin and ferulic acid esterase from solid koji:

[0077] Step one, koji extraction

[0078] Put the fermented yeast koji in the extraction tank, and add phosphate buffer solution with a concentration of 0.3mol / L and pH 6.5, which is 4.5 times the volume of the yeast koji, fully stir at 35°C for 25 minutes, and stand at the same temperature for 40 minutes; Separation of the extraction solution, adding 2.5 times the volume of fermented koji with a concentration of 0.3mol / L, pH 6.5 phosphate buffer solution, fully stirring for 15 minutes, standing still for 35 minutes for the second extraction, mixing the two extraction solutions, and then passing through the plate The frame filter removes impurities after filtering;

[0079] Step 2: Separation of bacteriocin and ferulic esterase by ultrafiltration

[0080] The pH of the extract was adjusted to 6.0 with hydrochloric acid or sodium hydroxide solution, and the bacteriocin and ferulic est...

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Abstract

The invention discloses a method for preparing bacteriocin and feruloyl esterase through solid-state fermentation of bacillus coagulans. The method comprises the following steps of step 1, preparing a liquid leavening agent; step 2, preparing a solid-state fermentation culture medium; step 3, solid state fermentation; step 4, separating and extracting bacteriocin and feruloyl esterase from a solid-state koji material; step 5, separating bacteriocin and feruloyl esterase by ultrafiltration; and step 6, spray drying preparation of feruloyl esterase and bacteriocin. The bacteriocin disclosed by the invention has a wide antibacterial spectrum, has a relatively strong inhibition effect on common pathogenic bacteria such as Listeria monocytogenes, bacillus cereus, staphylococcus aureus and other gram-positive bacteria, and can inhibit gram-negative bacteria including salmonella paratyphi, pseudomonas aeruginosa, shigella dysenteriae and escherichia coli; and the coarse material culture medium is adopted as a fermentation raw material, the process is simple, the enzyme activity is high, the production cost of unit enzyme activity is low, and low-cost large-scale production of feruloyl esterase is facilitated.

Description

technical field [0001] The invention relates to a method for preparing bacteriocin and ferulic esterase by solid-state fermentation of bacillus coagulans, belonging to the technical field of biological preparations. Background technique [0002] Antibiotics are often used in the treatment or prophylaxis of food animals, or to promote growth (feed additives). The misuse and inappropriate use of antibiotics in the food and feed industry has led to the emergence of antibiotic-resistant strains, posing a serious threat to human health. Therefore, there is an urgent need to produce safe and effective new antibacterial substances to replace antibiotics used in the feed industry. In recent years, bacteriocins have attracted much attention because they can be used not only as feed additives but also as alternatives to antibiotics. Bacteriocins are ribosome-synthesized antimicrobial peptides that are not lethal to producing cells. These polypeptides are generally active against sp...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/18C12P21/02C07K14/32C07K1/34C12R1/07
CPCC07K14/32C12N1/20C12N9/18C12Y301/01073
Inventor 张娜丁飞鸿陈腾张传丽高兆建宋玉林耿云龙赵宜峰焦魏张建萍
Owner XUZHOU UNIV OF TECH
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