Method for producing haematochrome by using Bacillus subtilis

A technology of Bacillus subtilis and red pigment, which is applied in the field of purification and refinement of crude red pigment products, can solve the problems of large demand and waste of raw materials, and achieve the effects of high pigment yield, easy operation and good reproducibility

Inactive Publication Date: 2015-02-11
XUZHOU UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional extraction method using red plants as raw materials has a huge demand for raw materials and is easy to cause waste
Most of the reported microorganisms used to produce red pigment are fungi, such as Monascus, but fungi are limited by growth conditions and time. In this regard, researchers have explored a way to use bacteria (such as Bacillus subtilis) to produce The production process of red pigment is researched in its liquid fermentation and its optimization of fermentation conditions and the extraction and purification of fermentation products, so as to provide a simple and feasible production method and purification method for the development of natural red pigment

Method used

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  • Method for producing haematochrome by using Bacillus subtilis
  • Method for producing haematochrome by using Bacillus subtilis
  • Method for producing haematochrome by using Bacillus subtilis

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1: Screening of high-yield red pigment strains

[0035] (1) Enrichment culture:

[0036]Put 1g of soil sample (collected in humus-rich soil) into a 250ml Erlenmeyer flask filled with 80ml of sterile water, add sterile glass beads, and vibrate in a shaker at 150r / min for 20min. Take 3-4ml of the shaken emulsion, add it into 100ml enrichment medium, and place it in a constant temperature shaker at 35°C with a rotation speed of 170r / min for 48h. Enrichment medium: 5ml bran extract, 0.3g beef extract, 1g peptone, 0.5g NaCl, K 2 HPO 4 0.1g, MgSO 4 ·7H 2 O0.02g, H 2 O95mL, pH7.2, sterilized at 121°C for 20min.

[0037] (2) Primary screening:

[0038] The enriched culture solution was diluted stepwise to 10 -4 、10 -5 、10 -6 , take 0.2ml and spread it on the primary screening solid medium plate, and when the plate is air-dried until there is no flowing liquid on the surface, cultivate it in a constant temperature incubator at 32-37°C for 30-36 hours, then l...

Embodiment 2

[0050] Example 2: Morphological characteristics and physiological and biochemical characteristics of high-yield red pigment strain HJ

[0051] Bacterial strain described in the present invention, after beef extract peptone culture medium plate culture 18h, observe bacterial colony shape, surface is rougher, edge is not neat, and bacterial colony color is light red, and Gram's staining is positive. The shape of the bacteria is short rod rod. The cell size is 0.65μm×2.7μm, can form endophytic spores, and the cell has motility; the cellulolysis test is negative; in the sugar fermentation experiment, the strain can use soluble starch, glucose, maltose and sucrose as the only In other physiological and biochemical tests, the strain was positive for contact enzymes and VP assays, could hydrolyze starch, liquefy gelatin, and utilize citrate without producing fluorescent pigments, and the methyl red test was positive. The specific morphological, physiological and biochemical characte...

Embodiment 3

[0055] Embodiment 3: Shake flask fermentation method produces red pigment

[0056] 1. Preparation of fermentation strains: Streak the pre-screened and preserved Bacillus subtilis strains on the beef extract peptone medium plate, and then place them upside down in a constant temperature incubator at 37°C for 15-24 hours for activation.

[0057] The configuration method of the solid medium is: beef extract 0.3g, peptone 1.0g, agar powder 1.5g, add distilled water to 100mL, pH7.2, 121°C, sterilize for 20min.

[0058] 2. Prepare the seed solution of Bacillus subtilis: pick a single colony from the activated strain in step 1, insert it into a 250mL Erlenmeyer flask with 30mL seed medium, and culture it on a shaker at 35-37°C for 18-25h , speed 160-200r / min.

[0059] The composition of the seed medium: peptone 1g, beef extract 0.5g, yeast powder 0.5g, NaCl 0.5g, K 2 HPO 4 0.02g, add distilled water to 100ml, pH6.8-7.2, sterilize at 121°C for 20min.

[0060] 3. Shake flask fermen...

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Abstract

The invention belongs to the field of microbial fermentation, and relates to a process method for preparing haematochrome by using Bacillus subtilis through liquid fermentation and a technique for purifying and refining haematochrome crude products. A strain used in the method provided by the invention is Bacillus subtilis HJ with a Preservation No. of CGMCC (China General Microbiological Culture Collection Center) No. 5701. The method for producing the haematochrome through fermentation, provided by the invention, comprises the following steps: (1) preparation of a seed solution, namely, carrying out activated cultivation on a strain Bacillus subtilis with a Preservation No. of CGMCC No. 5701 so as to obtain the seed solution; (2) inoculating the seed solution to a fermentation medium to carry out fermentation, wherein the fermentation refers to shaking flask fermentation or deep liquid ventilating fermentation; and (3) extracting the haematochrome in Bacillus subtilis fermentation liquor. The yield of the haematochrome prepared in a shaking flask fermentation mode reaches more than 1.8 g / L. The yield of the haematochrome prepared in a deep liquid ventilating fermentation mode is 2.4 g / L.

Description

Technical field: [0001] The invention relates to the field of microbial fermentation, in particular to a process for preparing red pigment by liquid fermentation of Bacillus subtilis and a purification technology for crude red pigment. Background technique: [0002] Natural pigments can not only be used as food additives for food dyeing, some of them also have important physiological health functions, and are widely used in the cosmetics industry, textile, clothing, and home textile industries. my country's huge natural pigment research and development team has enabled my country's food industry to develop rapidly since the reform and opening up. After years of hard work, my country's food coloring industry has risen and formed a certain scale, becoming an indispensable industry in the future food industry. The traditional production of natural pigments from animals and plants has many disadvantages, but the use of microbial resources to produce natural pigments has low cos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P1/04C12R1/125
Inventor 高兆建杜永凯唐仕荣张建萍陈尚龙王东星张从省周刚
Owner XUZHOU UNIV OF TECH
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