A kind of method of producing propionibacterium bacteriocin by high-density fermentation

A technology of high-density fermentation and propionibacterium, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of relatively little research on nitrogen sources, and achieve the advantages of convenient transportation, industrial production, and increased production Effect

Active Publication Date: 2020-05-19
CHANGSHU INSTITUTE OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, a large number of studies are focused on the carbon source required for the growth of propionibacterium, and there are relatively few studies on nitrogen sources.

Method used

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  • A kind of method of producing propionibacterium bacteriocin by high-density fermentation
  • A kind of method of producing propionibacterium bacteriocin by high-density fermentation
  • A kind of method of producing propionibacterium bacteriocin by high-density fermentation

Examples

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Effect test

preparation example Construction

[0031] Preparation of indicator bacteria: Inoculate the activated E. coli into LB liquid medium, culture at 37°C and shake at 220r / min until the OD of the bacterial solution 600nm 0.2, and then diluted 100 times with sterile LB medium for use.

[0032] Preparation of distiller's grain supernatant: Distiller's grains were stirred at 30°C for 3 hours according to the mass ratio of distiller's grains to distilled water of 1:4, centrifuged in a centrifuge at 4200r / min for 5min and filtered to obtain distiller's grain filtrate.

[0033] Configuration of yeast solution: Dissolve 2g of yeast powder in 1000mL of distilled water to make 2g / L yeast solution.

[0034] The following examples carry out the mensuration of antibacterial activity according to the following method: fermented liquid is centrifuged 15min under the condition of 4 ℃, 6000r / min, gets supernatant, is adjusted to pH7.0 with 10% NaOH, then with 10% tartaric acid Adjust the pH to 5.5, and then perform ultrafiltration ...

Embodiment 1

[0036] 1. Seed culture: inoculate a single colony of Propionibacterium freudenreichii CS1420 into a modified PYG medium for activation, and culture anaerobically at 30°C for 2.5 days. Improved PYG medium formula: casein peptone 5.0g, peptone 5.0g, yeast extract 10.0g, beef extract 5.0g, glucose 5.0g, K2HPO4 2.0g, Tween 80 1.0ml, resazurin 1.0ml, saline solution ( Recipe attached below) 40.0ml, distilled water 950.0ml, hemin solution (recipe attached below) 10.0ml, vitamin K1 solution (recipe attached below) 0.2ml, L-cysteine ​​0.5g, pH 7.2. Salt solution: CaCl 2 2H 2 O 0.25g, MgSO 4 ·7H 2 O 0.5g, K 2 HPO 4 1.0g, KH 2 PO 4 1.0g, NaHCO 3 10.0g, NaCl 2.0g, distilled water 1.0L. Hemin solution: 50.0 mg of hemin, 1.0 ml of 1N NaOH, and 99.0 ml of distilled water. Vitamin K1 solution: vitamin K10.1ml, 95% ethanol 20.0ml.

[0037] 2. Expanded cultivation: transfer the seed solution obtained in the above step 1 into a Erlenmeyer flask equipped with SLB medium according t...

Embodiment 2

[0041] Same as Example 1, the difference is that the added concentration of carbon source is 2.5g / L.

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Abstract

The invention discloses a method for producing propionibacterium bacteriocin through high-density fermentation. The method comprises the following steps: 1) performing seed cultivation; 2) performingenlargement cultivation; and 3) inoculating the seed liquid subjected to enlargement cultivation in the step (2) into a 5 L fermentation tank with a 2 L SLB culture medium according to 10 percent inoculation quantity, performing shaking table fermentation at the fermentation temperature of 30 DEG C and the shaking table rotating speed of 100 rpm, introducing CO2 during fermentation to maintain thetank pressure to be 0.05 Mpa and control the pH to be 6.0, supplementing a carbon source and a nitrogen source separately every other 24 hours starting from the 96th hour of fermentation and continuously supplementing for 5 times, wherein 100 mL of carbon source and 100 mL of nitrogen source are supplemented separately each time, the carbon source is a mixed carbon source comprising 75 percent ofsodium lactate and 25 percent of glucose, the adding concentration of the carbon source is 2 to 4 g / L, the nitrogen source comprises a yeast solution and corn vinasse clear liquid, the volume ratio of the yeast solution to the corn vinasse clear liquid is 1:(10-50), and the mass ratio of the vinasse to the distilled water in the corn vinasse clear liquid is 1:4. Due to supplementation in batches,high-density, high-yield and high-concentration cultivation of propionibacterium is guaranteed.

Description

technical field [0001] The invention relates to a production method of Propionibacterium bacteriocin, in particular to a method for producing Propionibacterium bacteriocin by high-density fermentation. Background technique [0002] As people pay more and more attention to food safety issues, the development and application of natural preservatives are widely valued. Among them, the microbial source preservative nisin (Nisin) in natural preservatives was approved by FAO / WHO as early as 1969 as an efficient and safe natural food preservative; ε-polylysine (ε-PL) was approved in 2003 In October, it was approved by the FDA as a safe food preservative. The United States, South Korea and Japan have allowed the application of ε-PL in food preservation and preservation; lysozyme has also been widely used in food preservation such as meat products, aquatic products and dairy products. In addition, microbial preservatives such as kojic acid, natamycin, methanotrophin, and reuterin al...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/00C12N1/20C12R1/01
CPCC12N1/20C12P21/00
Inventor 郑丽雪王立梅齐斌王家皓徐田甜
Owner CHANGSHU INSTITUTE OF TECHNOLOGY
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