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High-efficiency preparation method of Iturin A and homologue of Iturin A

A technology of iturin and its homologues, applied in the field of microorganisms, can solve the problems of increasing the difficulty of fermentation process, high production cost, and difficulty in realizing industrialized production, and achieves the effects of reducing foam, increasing yield and increasing concentration

Inactive Publication Date: 2015-06-10
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It carried out a 5L tank fermentation study. As a result, in the fermentation broth of the wild strain SD142, the yield of Iturin A and its homologues reached 3.8g / L; The yield reached 6.7g / L; but the production cost of this method is still on the high side, and with the increase of nitrogen source, there are disadvantages such as serious foaming in fermentation and increasing the difficulty of fermentation process, so it is difficult to realize industrial production

Method used

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  • High-efficiency preparation method of Iturin A and homologue of Iturin A
  • High-efficiency preparation method of Iturin A and homologue of Iturin A
  • High-efficiency preparation method of Iturin A and homologue of Iturin A

Examples

Experimental program
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Effect test

Embodiment 1

[0070] Use 12 1000mL triangular flasks, each containing 300mL medium A (soluble starch 1.0% glycerin 0.6%, glucose 5.0%, peptone 1.0%, yeast extract 0.5%, magnesium sulfate 0.5%, potassium dihydrogen phosphate 0.5%, sodium chloride 0.3%), sterilized at 120°C for 30 minutes, inoculated with activated Bacillus subtilis (Bacillus subtilis) ZK-H28 bacterial solution after cooling, placed at 30°C, and shaken the flask for 18 hours. Inoculate the cultured strain solution into a 100L fermenter with 50L sterilized medium B (medium B: corn flour 2.0%, maltose 1.0%, sucrose 0.5%, glycerol 0.5%, glucose 1.0%, peptone 1.0%, soybean flour hydrolyzate 5.0%, yeast extract 0.5%, fish bone meal 1.0%, potassium dihydrogen phosphate 0.5%, ammonium nitrate 0.5%, magnesium sulfate 0.2%, sodium chloride 0.3% ), at a temperature of 28°C-30°C, after aeration and stirring for 5-15 hours, the amino acid feeding solution (g / mL) (Lys1.0x10 -5 , Ser1.0x10 -4 , Gln2.0x10 -4 ,Asp5.0x10 -4 , Asn1.0x10 -...

Embodiment 2

[0075] Use 10 1000mL triangular flasks, each containing 300mL medium A (soluble starch 2.0%, glycerin 0.5%, glucose 5.0%, peptone 3.0%, yeast extract 0.3%, magnesium sulfate 0.6%, potassium dihydrogen phosphate 0.6%, chloride Sodium 0.4%), sterilized at 120°C for 30 minutes, inoculated with activated Bacillus subtilis (Bacillus subtilis) ZK-H6 (CGMCC No.2897) bacteria solution after cooling, placed at 30°C, shake flask cultured for 18 hours . Inoculate the cultured strain solution into a 100L fermenter with 50L sterilized medium B (medium B: corn flour 2.0%, maltose 1.0%, sucrose 0.5%, glycerol 0.5% %, glucose 1.0%, peptone 1.0%, soybean flour hydrolyzate 5.0%, yeast extract 0.5%, fish bone meal 1.0%, potassium dihydrogen phosphate 0.5%, ammonium nitrate 0.5%, magnesium sulfate 0.2%, sodium chloride 0.3%) , at a temperature of 28°C-30°C, after aeration and stirring for 5-15 hours, the amino acid feeding solution (g / mL) (Lys 1.0x10 -5 , Ser 1.0x10 -4 , Gln 2.0x10 -4 , Asp 5...

Embodiment 3

[0080] Use 12 1000mL triangular flasks, each containing 300mL medium A (soluble starch 4.0%, glycerin 1.0%, glucose 3.0%, peptone 1.0%, yeast extract 0.5%, magnesium sulfate 0.5%, potassium dihydrogen phosphate 0.5%, chloride Sodium 0.3%), sterilized at 120°C for 30 minutes, inoculated with activated Bacillus subtilis ZK-H28 bacterial solution after cooling, placed at 30°C, and shaken the flask for 18 hours. Inoculate the cultured strain solution into a 100L fermenter with 50L sterilized medium B (medium B: 3.0% corn flour, 1.0% maltose, 1.5% sucrose, glucose 1.0%, peptone 2.0%, soybean flour hydrolyzate 4.0%, yeast extract 0.5%, fish bone meal 1.0%, potassium dihydrogen phosphate 0.5%, ammonium nitrate 0.5%, magnesium sulfate 0.2%, sodium chloride 0.3%), at 28 ℃-30℃, aeration and stirring for 3-15 hours after fermentation, add amino acid feeding solution (g / mL) intermittently (Lys 0.5x10 -5 , Ser 1.5x10 -4 , Gln5.0x10 -5 , Asp 5.0x10 -4 , Asn 2.0x10 -4 , Glu 7.0x10 -5 ,...

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Abstract

The invention belongs to the technical field of microbes and particularly relates to a high-efficiency preparation method of Iturin A and a homologue of Iturin A. The method comprises the following steps: culturing a Bacillus strain and a genetically modified bacterium thereof in a primary fluid medium, and then inoculating a secondary fluid medium with the primary fluid medium as a seed solution, wherein the strain can generate Iturin A and the homologue of Iturin A; after inoculating the secondary fluid medium with the seed solution, performing fermentation culture and the feed-batch culture of amino acid and the like; and collecting Iturin A and the homologue thereof from a fermentation culture solution after fermentation is over. The method provided by the invention has the characteristics of high production efficiency, easy industrialization and the like.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a method for efficiently preparing iturin A (Iturin A) and its homologues. Background technique [0002] Iturin A (Iturin A) and its homologues are mainly derived from secondary metabolites of Bacillus strains such as Bacillus.subtilis, which have broad-spectrum disease resistance and are not easy to develop drug resistance And other advantages, it is considered to be a natural antibacterial substance with high efficiency to inhibit plant and animal pathogenic bacteria. In particular, Iturin A has strong antifungal properties and also inhibits the activity of some bacteria. [0003] Usually Bacillus (Bacillus) strains such as Bacillus subtilis (B.subtilis) produce two types of secondary metabolites, one is biosurfactin (Surfactin) with biosurfactant function, and the other is antifungal activity Iturin A and its homologues. [0004] Iturin substances are a m...

Claims

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Application Information

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IPC IPC(8): C12P21/04C12R1/07C12R1/125C12R1/10
Inventor 谭红钟娟周金燕彭文璟杨杰肖亮张智舒丹罗笛
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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