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A prodigiosin-producing bacterium and its production method and application

A technology for prodigiosin and bacteria to produce prodigiosin is applied in the field of microbial fermentation, and can solve the problems of lowering prodigiosin fermentation production efficiency, difficulty in realizing continuous prodigiosin fermentation, large amount of organic solvent, and the like. Improved fermentation yield, good inhibitory effect, and short fermentation period

Active Publication Date: 2021-03-26
XUZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above steps need to consume a large amount of organic solvent, complicated operation and high energy consumption
The use of organic solvents will not only pollute the environment, but also kill the bacteria, making it difficult to realize the continuous fermentation of prodigiosin and reducing the fermentation production efficiency of prodigiosin

Method used

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  • A prodigiosin-producing bacterium and its production method and application
  • A prodigiosin-producing bacterium and its production method and application
  • A prodigiosin-producing bacterium and its production method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Isolation and identification of the bacterial strain BWL1001 of embodiment 1

[0028] 1.1 Isolation of strain BWL1001

[0029] Soil samples were collected from farmland in Tongshan District, Xuzhou City, Jiangsu Province.

[0030] The collected soil samples were gradually diluted with 0.9% sterile saline, and then the dilutions were sequentially spread on the R2A solid medium (the composition of the R2A solid medium was as follows: soluble starch 0.5 g / L, glucose 0.5 g / L , yeast powder 0.5 g / L, tryptone 0.5 g / L, acid hydrolyzed casein 0.5 g / L, MgSO 4 •7H 2 O 0.05 g / L, K 2 HPO 4 0.3g / L, agar powder 20 g / L), put the coated medium upside down and put it in a constant temperature incubator at 28°C, and observe a red colony after culturing for 24 hours. After the strain was purified by the four-section line separation method, it was stored in a -80°C refrigerator with 20% glycerol solution.

[0031] 1.2 Strain identification

[0032] (1) Morphological identification ...

Embodiment 2

[0035] Example 2 Transformation of soybean oil production and in-situ extraction of prodigiosin

[0036] The specific implementation is as follows:

[0037] Step 1, preparation of seed solution: strain Serratia marcescens BWL1001 was inoculated into the seed liquid medium, and the components of the seed liquid medium were as follows: peptone 10 g / L, sodium chloride 10 g / L, magnesium sulfate heptahydrate 0.5 g / L, dipotassium hydrogen phosphate 0.3 g / L; 28°C, 200 rpm shaker constant temperature culture for 12-16 h;

[0038] Step 2, production of prodigiosin by fermentation: inoculate the seed liquid obtained in step 1 into the fermentation medium at a ratio of 1% (v / v), and the components of the fermentation medium are as follows: peptone 10 g / L, sodium chloride 10 g / L, magnesium sulfate heptahydrate 0.5 g / L, dipotassium hydrogen phosphate 0.3 g / L, soybean oil 10% (w / v); then 28°C, 200 rpm shaker constant temperature fermentation culture for 24-48 hours;

[0039] Step 3, in-...

Embodiment 3

[0043] The prodigiosin product obtained by fermentation was used for algal inhibition experiment analysis, in which Microcystis aeruginosa ( Microcystis aeruginosa ) were cultured in TAP liquid medium, and different concentrations (0.1 μg / mL, 0.2 μg / mL, 0.5 μg / mL, 1 μg / mL, 3 μg / mL) of prodigiosin were added, and after 48 h, the The concentration of algal cells in the culture medium. The calculation method of algae inhibition rate is as follows: (C0-Cp) / C0*100%, where C0 represents the concentration of algae cells in the culture medium without adding prodigiosin; concentration of algal cells.

[0044] The result is as Figure 4 As shown, it can be seen that the prodigiosin showed a good inhibitory effect on the growth of Microcystis aeruginosa, and when the product dosage of prodigiosin reached more than 1 μg / mL, Microcystis aeruginosa ( Microcystis aeruginosa ) has an algae inhibition rate of more than 80%, indicating that the prodigiosin product can be used to prepare ...

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Abstract

The invention discloses a prodigiosin-producing bacterium and its production method and application. The prodigiosin-producing bacterium is Serratia marcescens BWL1001, the preservation number is CGMCC No.18953, and it produces prodigiosin The method comprises: the preparation of seed liquid; the seed liquid is transferred to the fermentation medium with soybean oil as carbon source by the ratio of 1% (v / v), 28 ℃ constant temperature shaker culture 24-48 hours obtains fermentation liquid Centrifuge the fermentation broth, collect the upper oil phase, add 1 to 3 times the volume of acidic methanol with pH=3 to the oil phase, oscillate and mix, let stand for 30 minutes, centrifuge, collect the acidic methanol phase, remove the solvent to obtain bleeding A red pigment product, the prodigiosin exhibits a good inhibitory effect on the growth of Microcystis aeruginosa. The bacterial strain used in the invention has fast growth rate, short fermentation cycle, high prodigiosin output and low production cost, and plays an important role in promoting the industrial application of prodigiosin.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a prodigiosin producing bacterium and its production method and application. Background technique [0002] Prodigiosin (Prodigiosin) is a natural red pigment containing a three-pyrrole ring structure, which has a variety of biological activities: it can selectively and effectively inhibit the growth of various cancer cells, and has weak toxicity to normal cells; Prodigiosin can effectively inhibit the rejection caused by organ transplantation, and can be used to treat autoimmune diseases; prodigiosin and its derivatives have a strong killing effect on malaria parasites and trypanosoma; they have broad-spectrum bactericidal effects, It can effectively kill a variety of harmful bacteria and fungi; prodigiosin has efficient anti-algae activity, can kill planktonic algae that cause red tides and water blooms, and shows great potential in water pollution control; in add...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P17/16A01N43/36A01P13/00C12R1/43
CPCA01N43/36C12P17/165C12N1/205C12R2001/43
Inventor 刘伟杰孙地刘聪田雁宁杨静朱静榕蒋虹
Owner XUZHOU NORMAL UNIVERSITY
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