Dedicated bacterial strains for producing indigo blue pigments

An indigo pigment and production technology, applied in microorganism-based methods, bacteria, microorganisms, etc., can solve the problems of long strain culture period, low pigment secretion capacity, unstable enzyme, etc., and achieve constant synthesis capacity, simple operation, and production. Efficient effect

Inactive Publication Date: 2013-06-12
XUZHOU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the reported microbial catalysis also has many problems in the production and application of indigo pigment: such as the long culture period of the strain, low pigment secretion ability, unstable enzyme in the reaction medium, etc. strains are significant

Method used

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  • Dedicated bacterial strains for producing indigo blue pigments
  • Dedicated bacterial strains for producing indigo blue pigments
  • Dedicated bacterial strains for producing indigo blue pigments

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The screening identification of embodiment 1 producing indigo pigment strain

[0030] 1. Sample collection

[0031] Soil samples were collected from the sludge near the sewage treatment plant in Xuzhou City, Jiangsu Province.

[0032] 2. Isolation and screening of bacterial strains On September 12, 2011, soil samples were collected from the sludge of Kuihe River, Xuzhou City, Jiangsu Province, and the soil samples were collected from Dagao Zhaojian.

[0033] Prepare solid primary screening medium (unit g / 100ml): yeast extract 0.2, NaCl 0.2, (NH 4 ) 2 SO 4 0.2, NH 4 NO 3 0.1,K 2 HPO 4 0.01, MgSO 4 ·7H 2 O 0.01, FeSO 4 ·7H 2 O0.004, agar powder 1.5, pH7.5. The prepared culture is based on 121°C, sterilized for 15 minutes, and poured onto the plate. Indole was made into an indole solution with a final concentration of 0.5g / 100ml with ethanol, and the sterilized medium was cooled to about 50°C, and 3ml of indole solution was added per 100ml of medium, mixed w...

Embodiment 2

[0039] The fermentation culture of embodiment 2 bacterial strains and the extraction of indigo pigment crude product

[0040] 1. Culture of strains

[0041] Prepare seed liquid medium (unit g / 100ml): peptone 1, glucose 0.5, yeast powder 0.5, NaCl 0.2, (NH 4 ) 2 SO 4 0.2, NH 4 NO 3 0.1,K 2 HPO 4 0.01, MgSO 4 ·7H 2 O 0.01, pH7.5. The prepared culture is based on 121°C and sterilized for 15 minutes. Inoculate a single colony of the strain JWDL into the above medium, fill a 250ml Erlenmeyer bottle with 40ml of liquid, and culture with shaking at 32°C and 180r / min for 20h.

[0042] Preparation of fermentation medium (g / L): peptone 0.5, glucose 0.5, yeast powder 0.5, (NH 4 ) 2 5O 4 0.25, MgSO 4 ·7H 2 O0.01, FeSO 4 ·7H 2 O0.04, NH 4 NO 3 0.1, pH7.5. The prepared culture is based on 121°C and sterilized for 15 minutes. Inoculate the cultured seed liquid into the sterilized fermentation medium according to the inoculum amount of 4%, fill the volume of liquid in ...

Embodiment 3

[0045] The structural identification of embodiment 3 pigment

[0046] 1. Thin-layer chromatography (TLC) analysis: the prepared crude indigo sample was dissolved in N,N-dimethylformamide (DMF). The sample and the standard sample were respectively spotted on the thin electrode of TLC. The developing solvent is methanol-acetone / 1:1 (v / v) solvent system. After fully unfolding, blow dry with a hair dryer. The result is as figure 2 As shown, a blue spot appears at the same position on the chromatographic plate.

[0047] 2. Ultraviolet-visible broad-spectrum scanning analysis: the indigo sample dissolved in N,N-dimethylformamide (DMF) is the same as the standard indigo sample, and the ultraviolet-visible (250nm-700nm) broad-spectrum scanning is performed. The result is as image 3 As shown, the prepared sample is basically consistent with the scanning broad-spectrum absorption curve of the standard sample, and both show absorption peaks at 285nm and 600nm. This proves that th...

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Abstract

The invention discloses dedicated bacterial strains for producing indigo blue pigments. The invention relates to provide bacillus subtilis bacterial strains for producing the indigo blue pigments. The manufacturing method includes the following steps: (1) bacterial strains fermental cultivation: after strains of the bacillus subtilis CGMCC NO.5698 are cultivated on LB solid medium with the temperature 30-37 Celsius degrees, a single colony is put into seed solution culture medium which are sterilized, the single colony is cultivated for 46-60 hours with the constant temperature of 26-34 Celsius degrees, the rotating speed of a shaker is 200r/min; the seed solution is put into liquid fermentation medium which is sterilized and then cultivated for 48-60 hours with the constant temperature, the rotating speed of the shaker is 200r/min; (2) picking up the bacillus subtilis indigo blue pigments. The dedicated bacterial strains for producing the indigo blue pigments have the advantages that the used bacillus subtilis bacterial strains are capable of efficiently combining indigo blue in the liquid state fermentation medium, production efficiency is high. Requirements of the dedicated bacterial strains are low to carbon sources and nitrogen sources, and genetic stability is good, and operation is simple.

Description

technical field [0001] The present invention relates to a Bacillus subtilis producing indigo pigment. Background technique: [0002] Indigo is a bright and stable blue pigment. Structurally, indigo is an aromatic compound mainly used for dyeing of fabrics, such as jeans are obtained by dyeing textiles with indigo. In addition to being used as a dye, indigo-indirubin (an indigo-like pigment) also has important applications in the pharmaceutical industry, and can have important therapeutic effects on some diseases, including cancer-related diseases, Alzheimer's disease, and delayed hypersensitivity. In the food industry, indigo can be added to food as a kind of food coloring additive. The traditional indigo production is mainly extracted from plants, and the leaves of some plants such as woad, Polygonum indigo, horse blue, and wild green tree are rich in indigo pigment. Indigo extracted from plants has high safety and less pollution, but its yield and purity are low, which ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/16C12R1/125
Inventor 高兆建纪伟陈宏伟候进慧孙会刚陈尚龙巫永华
Owner XUZHOU UNIV OF TECH
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