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Prodigiosin (PG) producing strain as well as preparation method and application thereof

A prodigiosin and bacteria-producing technology, applied in the field of bioengineering, can solve the problems of lack of high-quality strains and long fermentation cycle, and achieve the effects of shortening the fermentation cycle, protecting activity, and increasing production

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

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a prodigiosin producing bacterium and its preparation method and application, 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 (PG) producing strain as well as preparation method and application thereof
  • Prodigiosin (PG) producing strain as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1: Screening and isolation of prodigiosin producing bacteria

[0024] Collect the seaside soil in Laishan District, Yantai City, Shandong Province, take 1g of the sample, add 100ml of sterile deionized water, apply the LB solid medium plate after gradient dilution, incubate at 28°C for 24h-48h, and screen the red colonies. A prodigiosin-producing strain P3 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; pH7.0~7.5, Autoclave at 121°C for 20 minutes, and when cooled to 50-60°C, invert the plates.

[0025] Observation on the plate showed that the colony was bright red, distributed in a corrugated shape, and Gram staining was negative. The bacterium was preserved in the General Microbiology Center CGMCC of the China Microbiological Culture Collection Management Committee on June...

Embodiment 2

[0027] Embodiment 2: Fermentation culture of prodigiosin producing bacteria P3

[0028]The seed culture medium adopts LB liquid culture medium, and its composition is that 1L culture medium contains: 10g tryptone, 5g yeast powder, 10g NaCl, the balance is water.

[0029] Take 10-100 μL of the prodigiosin-producing bacteria P3 concentration of 50-70wt% glycerol bacteria solution stored at -80°C, add it to 5-10mL LB liquid medium, and culture at 28-30°C for 12-24h to OD 600 0.6 to 1.0, use this as the seed solution for fermentation.

[0030] Fermentation medium adopts improved inorganic salt medium, 1L medium contains: sodium chloride (NaCl) 0.5 ~ 1g, potassium dihydrogen phosphate (KH 2 PO 4 ) 1g, dipotassium hydrogen phosphate (K 2 HPO 4 ) 3g, ammonium chloride (NH 4 Cl) 1~1.2g, magnesium sulfate (MgSO 4 ) 1g, ammonium nitrate (NH 4 HO 3 ) 2g, fructose or sucrose 20-50g, yeast powder 0.1-0.5g, the balance is water, pH7.0-8.0.

[0031] The seed liquid is inoculated int...

Embodiment 3

[0032] Embodiment 3: Preparation of prodigiosin P3PG

[0033] The fermented liquid described in Example 2 was respectively placed in a centrifuge tube at a temperature of 4-10° C., a rotation speed of 6000-10000 rpm, and centrifugation for 8-10 minutes. Add 1:1 to 1:2 (v / v) ethyl acetate to the obtained supernatant, mix well and let it stand for 10 to 20 minutes, use a separatory funnel to separate and save the organic phase; add 3 to 5 times the volume of Acetone, mix thoroughly, let stand for 5-10min, centrifuge at 6000-8000rpm for 3-5min at 4-10°C, and take the organic phase. The obtained ethyl acetate and acetone extract was distilled under reduced pressure at 50-60°C to 1-2% of the total volume, and freeze-dried to obtain the crude prodigiosin P3PG.

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Abstract

The invention relates to fermentation production and preparation of an active product, belongs to the field of bioengineering and in particular relates to a prodigiosin (PG) producing strain as well as a preparation method and an application thereof. The PG producing strain is Proteus Vulgaris and is preserved on June 24th, 2013 in CGMCC (China General Microbiological Culture Collection Center) and has the preservation number of CGMCCNo.7815 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 PG producing strain can produce PG through fermentation, and an improved inorganic salt culture medium is adopted and is cultured for 18-24 hours at a culture temperature of 20-28 DEG C and a rotating speed of 150-300rpm, and yield of PG can be 80-120mg / L. The produced prodigiosin has an inhibitory activity to staphylococcus aureus, salmonella paratyphi, tritirachium album and pseudomonas aeruginosa and has a good application prospect in bio-preparation of the PG.

Description

technical field [0001] The invention relates to the fermentation production and preparation of active products, belongs to the field of bioengineering, and specifically relates to a prodigiosin-producing bacterium and a preparation method and application thereof. Background technique [0002] Prodigiosins (PG) is a general term for a class of natural red pigments with a three-pyrrole ring, also known as prodigiosin and prodigiosin, with a molecular formula of C 20 h 25 N 3 O (M=323.1968), is a kind of fat-soluble red pigment. PG solid is a dark red pigment with green reflection, melting point 151-152°C, almost insoluble in water, easily soluble in organic solvents such as methanol, ethanol, ethyl acetate, acetone, chloroform and benzene. It is orange-yellow in alkaline or neutral solution, and red in acidic solution. Under different acid-base conditions, it has a unique maximum ultraviolet absorption wavelength: acidic conditions have characteristic absorption peaks betw...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/16C12R1/37
Inventor 焦绪栋秦松崔玉琳李莉莉
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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