Prodigiosin producing strain and method for producing prodigiosin through fermentation of prodigiosin producing strain

A prodigiosin and bacteria-producing technology, which is applied in the field of fermentation engineering, can solve the problems of long fermentation cycle and lack of high-quality strains of active prodigiosin.

Active Publication Date: 2013-12-11
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a prodigiosin-producing bacterium and a method for fermenting and producing prodigiosin, so as to solve the problems of lack of high-quality strains and long fermentation period in the fermentation preparation of active prodigiosin

Method used

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  • Prodigiosin producing strain and method for producing prodigiosin through fermentation of prodigiosin producing strain
  • Prodigiosin producing strain and method for producing prodigiosin through fermentation of prodigiosin producing strain
  • Prodigiosin producing strain and method for producing prodigiosin through fermentation of prodigiosin producing strain

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Embodiment 1

[0025] Example 1: Screening and isolation of prodigiosin producing bacteria

[0026] Collect the soil in the coastal area of ​​Laishan District, Yantai City, Shandong Province, take 1g of the sample, add 100ml of sterile deionized water, apply LB solid medium plate after gradient dilution, incubate at 28°C for 24h to 48h, and screen red colonies. A prodigiosin-producing strain P11 was obtained after repeated domestication, separation and purification. Among them, the composition of LB solid medium is: tryptone (Tryptone) 10g / L, yeast powder (Yeast extract) 5g / L, sodium chloride (NaCl) 10g / L, the balance is water; pH7.0~8.0, Autoclave at 121°C for 20 minutes, and when cooled to 50-60°C, invert the plates.

[0027] Observed on the LB solid medium plate, the bacterial colony is red, distributed in a circular shape, with a moist surface, and Gram staining is negative. The bacterium was preserved in CGMCC, General Microorganism Center of China Committee for Culture Collection of ...

Embodiment 2

[0029] Embodiment 2: the cultivation of P11 seed liquid

[0030]The seed medium adopts LB liquid medium, and its composition is that 1L medium contains: trypton (trypton) 10g, yeast powder (yeast extract) 5g, sodium chloride (NaCl) 10g, the balance is water, pH7.0~ 8.0. Take 10-100 μL of the prodigiosin-producing bacterium P11 stored at -80°C with a concentration of 50-70wt% glycerin bacteria and inoculate it into 5-10 mL of seed medium. Culture conditions: 28-30°C, 150-250 rpm (rev / min), culture time 12-24 hours, until OD600nm reaches 0.6-1.2, observe the production of pigment with naked eyes, and obtain seed liquid.

Embodiment 3

[0031] Embodiment 3: the fermentation condition optimization of P11

[0032] The fermentation medium adopts the improved inorganic salt medium. 1L medium contains: Sodium chloride (NaCl) 0.05g Potassium dihydrogen phosphate (KH 2 PO 4 ) 1g, dipotassium hydrogen phosphate (K 2 HPO 4 ) 3g, ammonium chloride (NH 4 Cl) 1 g, magnesium sulfate (MgSO 4 ) 1g, ammonium nitrate (NH 4 HO 3 ) 2g, fructose or glucose 20g, yeast powder 0.5g, the rest is water, pH8.0.

[0033] Culture conditions: Inoculate the seed liquid obtained through seed medium cultivation into the fermentation medium at 0.5-2wt%, avoid light, cultivate at a temperature of 24-28°C, rotate at a speed of 250-360rpm, and cultivate for 20-26 hours. until the bacterial liquid produces obvious red color ( figure 1 ). Measure the OD600nm absorbance value of the bacterial liquid at an interval of 1 to 2 hours. At the same time, take an appropriate amount of the bacterial liquid and centrifuge it at 10000 to 12000 rpm...

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Abstract

The invention belongs to the technical field of fermentation engineering and in particular relates to a prodigiosin producing strain and a method for producing prodigiosin through fermentation of the prodigiosin producing strain. The prodigiosin producing strain is acid producing klebsiella (Klebsiella oxytoca), is preserved on June 24th, 2013 in CGMCC (China General Microbiological Culture Collection Center) and has the preservation number of CGMCCNo.7816 and the preservation unit address of No.3 No.1 Courtyard, West Beichen Road, Chaoyang District, Beijing, Institute of Microbiology of Chinese Academy of Sciences. The prodigiosin producing strain can produce the prodigiosin through fermentation, an improved inorganic salt culture medium is adopted and is cultured away from light for 20-26 hours at a temperature of 24-28 DEG C and a rotating speed of 250-360rpm, and pigment yield can be 70-90mg/L. The produced prodigiosin is sensitive to natural light and ultraviolet rays and has an inhibitory activity to staphylococcus aureus, salmonella paratyphi and tritirachium album. The prodigiosin producing strain has a good application prospect in fermentation production of the prodigiosin and the fields of tumour suppression, food, cosmetics and the like.

Description

technical field [0001] The invention belongs to the technical field of fermentation engineering, in particular to a prodigiosin producing bacterium and a method for producing prodigiosin by fermentation. Background technique [0002] Prodigiosins (PG) is a general term for a class of natural red pigments with a tripyrrole ring structure. Current research shows that this kind of pigment generally has antibacterial, antitumor and immunosuppressive activities, and has good application prospects in industrial dyeing and red tide suppression. Especially in the aspect of anti-tumor, because of its specificity against cancerous tissue and low toxicity to normal cells, it has become a hot spot in anti-tumor research. The rapid and efficient preparation of active prodigiosin is one of the bottlenecks restricting its large-scale research and application. [0003] Prodigiosin is a metabolic by-product of some bacteria or Streptomyces. Most of its producing bacteria are isolated from...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/16C12R1/22
Inventor 焦绪栋秦松李莉莉
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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