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New method for preparing Iturin A and homolugues thereof

A technology of iturin and homologues, applied in the field of microorganisms, can solve the problems of difficulty in realizing industrial production, high production costs, and increasing the difficulty of fermentation process, and achieve the effects of increasing production, increasing concentration, and reducing foam

Inactive Publication Date: 2010-09-15
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production cost of this method is still relatively high, and there are disadvantages such as severe foaming in fermentation and increased difficulty of fermentation process with the increase of nitrogen source, so it is difficult to realize industrial production

Method used

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  • New method for preparing Iturin A and homolugues thereof
  • New method for preparing Iturin A and homolugues thereof
  • New method for preparing Iturin A and homolugues thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Use 10 1000mL Erlenmeyer flasks, each containing 300mL medium A (glucose 2.0%, peptone 3.0%, beef extract 0.5%, magnesium sulfate 0.35%, potassium dihydrogen phosphate 0.3%), sterilize at 120°C for 30 minutes, after cooling The activated Bacillus subtilis ZK-H6 bacterial solution was inoculated, placed at a temperature of 30° C., and shaken for 18 hours. Inoculate the cultured strain solution into 50L sterilized medium B, 10g / L adsorption immobilized cell material (polylactic acid microbody, or macromolecular material microcapsules such as PVA, or Sawdust) and 2% viscous agent (dextrin, or xanthan gum, or sodium alginate, or sodium carboxymethylcellulose (CMC)) in a 100L fermenter, (medium B: starch 2.0% lactose 1.0% sucrose 1.0% glycerol 0.5% glucose 1.0% peptone 1.0% peanut flour 5.0% soybean flour hydrolyzate 2.0% biological nitrogen 3.0% potassium dihydrogen phosphate 0.1% sodium chloride 0.5% iron sulfate 0.5%). At a temperature of 28°C-30°C, after 8-15 hours of a...

Embodiment 2

[0064]Use 12 1000mL Erlenmeyer flasks, each containing 300mL medium A (glucose 2.0%, peptone 3.0%, beef extract 0.5%, magnesium sulfate 0.35%, potassium dihydrogen phosphate 0.3%), sterilize at 120°C for 30 minutes, after cooling The activated Bacillus subtilis ZK-H6 bacterial solution was inoculated, placed at a temperature of 30° C., and shaken for 18 hours. Inoculate the cultured bacterial seed liquid into 50L sterilized medium B, 15g / L adsorption immobilized cell material (natural fiber material, or polylactic acid microbody, or sawdust) and 5 % viscous agent (agar, or soluble starch, or sodium alginate, or sodium carboxymethylcellulose (CMC)) in a 100L fermenter, (medium B: starch 3.0%, lactose 2.0%, glycerol 0.6%, molasses 3.0% Glucose 1.0%, Peptone 0.1%, Peanut flour hydrolyzate 5.0%, Soybean flour hydrolyzate 2.0%, Biological nitrogen 2.0%, Potassium dihydrogen phosphate 0.1%, Sodium chloride 0.5%, Iron sulfate 0.5%), at 28°C-30°C, aeration After stirring and fermenti...

Embodiment 3

[0069] Use 15 1000mL Erlenmeyer flasks, each containing 300mL medium A (glucose 3.0%, peptone 3.0%, beef extract 0.8%, magnesium sulfate 0.35%, potassium dihydrogen phosphate 0.3%), sterilize at 120°C for 30 minutes, after cooling The activated Bacillus subtilis ZK-H6 bacterial solution was inoculated, placed at a temperature of 30° C., and shaken for 18 hours. Inoculate the cultured strain solution into 50L sterilized medium B, 30g / L adsorption immobilized cell material (polylactic acid microbody, or macromolecular material microcapsules such as PVA, or Corn cob, or sawdust) and 8% viscous agent (soluble starch, or dextrin, or sodium alginate) in a 100L fermenter, (medium B: starch 2.0%, lactose 2.5%, glycerin 1.5%, waste molasses 3.0%, glucose 1.0 % peptone 0.5%, peanut powder 5.0%, soybean powder 2.0%, biological nitrogen 4.0%, potassium dihydrogen phosphate 0.1%, sodium chloride 0.5%, iron sulfate 0.5%), at 28°C-30°C, aerated and stirred for 10-15 hours Afterwards, medium...

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Abstract

The invention belongs to the technical field of microorganisms, particularly relates to a method for preparing Iturin A and homolugues thereof. The method comprises the following steps of: culturing bacillus strains and genetic improved strains thereof into a first-stage liquid culture medium, wherein the bacillus strains and genetic improved strains thereof are capable of producing the Iturin A and homolugues thereof; then, using as seed liquid and inoculating into a second-stage liquid culture medium; adding a certain proportion of adsorption and fixing cell material and thickening agent into the second-stage liquid culture medium before inoculating the seed liquid; and carrying out fermentation culture and feeding and supplying culture on the second-stage liquid culture medium after inoculating the seed liquid; and finally collecting the Iturin A and homolugues thereof from the fermentation culture liquid after the fermentation is finished. The invention has the advantages of simple process, convenient operation, high yield, easy industrialization, and the like.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a preparation method of iturin A (Iturin A) and its homologues. Background technique [0002] Iturin A (Iturin A) and its homologues are mainly derived from the secondary metabolites of Bacillus strains such as Bacillus subtilis, which have broad-spectrum disease resistance and are not easy to produce 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 (Iturin A) and its homologues. [0004] Iturin is a mixture o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04C12R1/07C12R1/10C12R1/125
Inventor 谭红钟娟周金燕李志东张晓勇杨杰肖亮付雯
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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