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Application of bacillus prodigiosus ITBB B5-1 to production of prodigiosins

A technology for the production of prodigiosin by Serratia marcescens, applied in the field of microorganisms, can solve the problems of low prodigiosin production and achieve broad application prospects

Active Publication Date: 2020-10-16
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the prodigiosin-producing Serratia marcescens strains reported so far have low prodigiosin production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: Serratia marcescens ITBB B5-1 of the present invention produces prodigiosin

[0039]1. Activate Serratia marcescens ITBB B5-1 in LB culture medium in the dark at 25°C for 2 days, take a single colony and inoculate it in a medium containing 100mL tryptone 15g / L, yeast extract 10g / L, sodium chloride 5g / L, pH value 7.0 liquid culture medium, shake culture at 250rpm under 25°C dark condition, cultivate for 12h, measure the OD value of the bacterial liquid under the condition of wavelength 600nm is 0.8, the bacterial liquid is used as the seed liquid.

[0040] 2. Inoculate the seed solution at a ratio of 3% in a liquid medium containing tryptone 15g / L, yeast extract 10g / L, sodium chloride 5g / L, and a pH value of 7.0. The liquid volume is 300mL / 1000mL. Shake flask culture was carried out at 250 rpm under dark conditions, the fermentation temperature was 22° C., and the fermentation time was 48 hours.

[0041] 3. Take 1mL of fermentation broth and centrifuge at ...

Embodiment 2

[0042] Embodiment 2: Serratia marcescens ITBB B5-1 of the present invention produces prodigiosin

[0043] 1. Activate Serratia marcescens ITBB B5-1 in LB culture medium in the dark at 25°C for 2 days, take a single colony and inoculate it in a medium containing 100mL tryptone 20g / L, yeast extract 12g / L, sodium chloride 5g / L, pH value 7.0 liquid culture medium, 25°C under dark conditions, shaking culture at 250rpm, culture for 15h, measure the OD of the bacterial solution 600 0.9 as the seed solution.

[0044] 2. Inoculate the seed solution at a ratio of 4% in a culture medium containing tryptone 20g / L, yeast extract 12g / L, sodium chloride 5g / L, and pH 7.0. Under the conditions of 250rpm shake flask culture, the fermentation temperature is 28°C, and the fermentation time is 72h.

[0045] 3. Take 1mL of fermentation broth and centrifuge at 12000g for 5min in a 1.5mL centrifuge tube, discard the supernatant, add 1mL of acidified ethanol to resuspend the cell pellet to extract p...

Embodiment 3

[0046] Embodiment 3: Serratia marcescens ITBB B5-1 of the present invention produces prodigiosin

[0047] 1. Activate Serratia marcescens ITBB B5-1 in LB culture solid medium under dark conditions at 25°C for 2 days, take a single colony and inoculate it in a medium containing 100mL tryptone 25g / L, yeast extract 15g / L, sodium chloride 5g / L, pH 7.0 liquid culture medium, 25°C under dark conditions, 250rpm shaking culture, culture to 18h, bacterial solution OD 600 The measurement was 1.0, and the bacterial liquid was used as the seed liquid.

[0048] 2. Inoculate the seed solution at a ratio of 5% in a culture medium containing tryptone 25g / L, yeast extract 15g / L, sodium chloride 5g / L, and pH 7.0. Under the conditions of 250rpm shake flask culture, the fermentation temperature is 25°C, and the fermentation time is 96h.

[0049] 3. Take 1mL of fermentation broth and centrifuge at 12000g for 5min in a 1.5mL centrifuge tube, discard the supernatant, add 1mL of acidified ethanol t...

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PUM

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Abstract

The invention relates to the field of microorganisms, in particular to an application of bacillus prodigiosus ITBB B5-1 to production of prodigiosins. The preservation No. of the bacillus prodigiosusis CGMCC No.7416. The prodigiosins are produced by the bacterial strain, the yield can achieve 9.8-15.5g in each liter of culture mediums, and the yield is notably higher than that of a bactria strainwhich is reported. The restriction of being low in yield of the prodigiosins by the bacillus prodigiosus bacterial strain is overcome, and the bacillus prodigiosus ITBB B5-1 has wide application prospects in the medicine markets.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to the application of Serratia marcescens ITBB B5-1 in the production of prodigiosin. Background technique [0002] Prodigiosins are a class of natural red pigments with a methoxypyrrole skeleton structure with 3 pyrrole rings, including Prodigiosin, Cycloprodigiosin, ten Undecylprodigiosin, Metacycloprodigiosin, Streptorubin B, etc. (You Zhongyu, Wang Yujie, Sun Shiqing, Liu Xiaoxia. Production of prodigiosin by microbial fermentation Research progress. Chinese Journal of Bioengineering, 2016, 32(10): 1332-1347). Among them, Prodigiosin has great potential application value in the medical field. Prodigiosin is an anticancer drug with high efficiency and low toxicity. Relevant studies have shown that the average half inhibitory concentration (IC 50 ) is 2.1umol / L, it has strong resistance to 60 kinds of cancer cell lines, and does not produce toxicity to normal cells. Prodigiosin ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/16C12N1/20C07D207/44C12R1/43
CPCC12P17/165C12N1/20C07D207/44Y02A50/30
Inventor 谭德冠张家明付莉莉马帅孙雪飘
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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