Prodigiosin high-producing strain and production method thereof

A technology for prodigiosin and its production method, which is applied to high-yield prodigiosin strains and its production field, can solve the problems of high cost, complex synthesis process, failure to implement large-scale chemical synthesis of Prodigiosin, etc., to reduce adverse effects, Effect of reducing energy consumption and noise

Active Publication Date: 2014-06-11
GUANGDONG PHARMA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In the 1960s, Rapoport et al. established the whole process of Prodigiosin chemical synthesis. In 1996, Alessio et al. proposed a new preparation method. However, due to the complex synthesis process and high cost, large-scale chemical synthesis of Prodigiosin has not been implemented.
However, after multiple rounds of screening, the high-yield strains were cultivated for more than 10 generations, and their pigment synthesis ability was found to be unstable or decreased.

Method used

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  • Prodigiosin high-producing strain and production method thereof
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  • Prodigiosin high-producing strain and production method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Isolation and identification of strains

[0035]1) Enrichment medium (g / L): tryptone 10, yeast extract 5, NaCl 7.5, phenol 100 mg, pH 7.0; inorganic salt medium (g / L): ammonium nitrate 1, dihydrogen phosphate Sodium 0.5, dipotassium hydrogen phosphate 0.5, magnesium sulfate 0.2, calcium chloride 0.1, sodium chloride 0.2, pH 7.0. Add 2% agar powder to the corresponding solid medium, and add an appropriate amount of phenol to the phenol-containing medium.

[0036] 2) Inoculate 5 mL of sewage into 45 mL of enriched medium containing phenol, and incubate at 30°C and 130 r / min for 72 h. Take 1 mL each of the acclimatized bacterial solution and spread it on the inorganic salt medium plate containing phenol and incubate for 72 h at 30°C. The final concentrations of phenol are 100 mg / L, 300 mg / L, 500 mg / L, 700 mg / L respectively , 1000 mg / L. The colonies with good growth conditions were picked and separated by streaking on the enrichment medium plate, cultured at 30°C for 72 ...

Embodiment 2

[0045] Embodiment 2 Batch fermentation mode

[0046] Primary shake flask culture (250 mL Erlenmeyer flask): sucrose 19 g / L, peptone 10 g / L, sodium chloride 3 g / L, magnesium sulfate 2.4 g / L, liquid volume 50 mL, shaking speed 130 rpm, The pH was 6, and cultured at 30°C for 20-24 h.

[0047] Secondary shake flask culture (2 L Erlenmeyer flask): 1%-5% peanut powder medium, liquid volume 50 mL, shaking speed 130 rpm, pH 6, culture at 30°C for 20-24 h.

[0048] Three-stage fermentation culture (50 L fermenter): 1%-5% peanut powder medium, loading coefficient 0.6, stirring speed 200 r / min, dissolved oxygen 20%-40%, pH 6, 5% inoculum size, Incubate at 25-30°C for 30-40 hours. The yield of prodigiosin reached 6-8 g / L.

Embodiment 3

[0049] Embodiment 3 Batch fermentation mode

[0050] (1) Seeds on slant: Take the slant of the test tube or the glycerol seeds frozen at -80°C, inoculate it on the slant of the LB test tube, and incubate in a biochemical incubator at 30°C for 20-24 h.

[0051] (2) According to the 5% inoculum amount, the three-stage culture method is used for amplified culture.

[0052] Primary shake flask culture (250 mL Erlenmeyer flask): sucrose 19 g / L, peptone 10 g / L, sodium chloride 3 g / L, magnesium sulfate 2.4 g / L, liquid volume 50 mL, shaking speed 130 rpm, The pH was 6, and cultured at 30°C for 20-24 h.

[0053] Secondary shake flask culture (2 L Erlenmeyer flask): sucrose 19 g / L, peptone 10 g / L, sodium chloride 3 g / L, magnesium sulfate 2.4 g / L, glycerol 5 g / L, liquid volume 50 mL , shaking speed 130 rpm, pH 6, cultured at 30°C for 20-24 h.

[0054] Tertiary fermentation culture (50 L fermenter): 19 g / L sucrose, 10 g / L peptone, 3 g / L sodium chloride, 2.4 g / L magnesium sulfate, 10 ...

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Abstract

The invention discloses a prodigiosin high-producing strain and a production method of the prodigiosin high-producing strain. The prodigiosin high-producing strain is named as serratiamarcescenszl3 and is stored in China Typical Culture Collection Center with a number of CCTCCNO: M201209 on April 4, 2012 in Wuhan University, Wuhan, China. The prodigiosin high-producing strain disclosed by the invention has the advantages of being fast in growth speed, stable in synthesis of prodigiosin, and high in expression index; after being cultured for 30 to 40 hours in 1 to 5% of a peanut powder culture medium on a 50L fermenting tank at 25 to 30 DEG C, the prodigiosin has a yield up to 6 to 8g / , and purity is more than 98%. With the adoption of a technology of fermenting in batch, fed-batch fermentation technical parameters, an optimized technology of agitating and digesting and a chromatography refine purifying technology provided by the invention, guidance is provided for industrial and scale production of the prodigiosin; and the prodigiosin can be applied to bio-pharmaceuticals and pharmaceutical chemical engineering fields.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a prodigiosin high-yielding bacterial strain and a production method thereof. Background technique [0002] Prodigiosins is a general term for a class of natural red pigments with a methoxypyrrole ring structure, which can be produced by a variety of actinomycetes and bacteria. Including prodigiosin (Prodigiosin) and its analogs and derivatives, such as Prodigiosin25-C, metacyclo Prodigiosin, Prodigiosen, cycloProdigiosin, desmethoxy prodigiosin and undecyl prodigiosin, etc. Prodigiosin and its derivatives can be used for dyeing textiles, killing algae that cause red tides and water blooms, and also have various biological activities such as antibacterial, antifungal, antimalarial, antitumor, and immunosuppressive. At present, people are most interested in the immunosuppressive activity of this substance and the effect of tumor cell apoptosis. For example, in 2007, the patented pro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P17/16C12R1/43
Inventor 周林朱爽
Owner GUANGDONG PHARMA UNIV
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