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Staphylococcus cohnii and method for preparing 5-aminolevulinic acid by using staphylococcus cohnii

A technology of aminolevulinic acid and staphylococcus, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of high cost, low yield, cumbersome chemical synthesis steps, etc., and achieve the effect of preventing decomposition

Inactive Publication Date: 2012-12-12
FUJIAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to cumbersome chemical synthesis steps, difficult to control by-products, low yield and other prominent problems, resulting in high cost and environmental pollution

Method used

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  • Staphylococcus cohnii and method for preparing 5-aminolevulinic acid by using staphylococcus cohnii
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  • Staphylococcus cohnii and method for preparing 5-aminolevulinic acid by using staphylococcus cohnii

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Experimental program
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Embodiment 1

[0019] Sampling was taken from different litter materials of a piggery in the northern suburbs of Fuzhou, each soil sample was weighed about 10 g, suspended in 0.9% NaCl solution, filtered with 8 layers of gauze to remove larger impurity particles, and inoculated to LB plates containing 50 ug / mL nystatin were cultured at 30°C for 24 hours, single colonies were selected, and single colonies were obtained by streaking on the LB plate, then D-glucose was used as the single carbon source for shake flask fermentation, and the fermentation broth was started for 12 hours. The fermentation broth was sampled every 3 hours, and the concentration of 5-aminolevulinic acid was quantitatively determined by spectrophotometry in the centrifuged supernatant. After preliminary screening, 4 strains of ALA-producing bacteria were isolated. Combined with the ALA production capacity, LA10 was finally selected as the starting strain.

[0020] Table 1 ALA production capacity of each strain

[0021]...

Embodiment 2

[0029] 1) with Staphylococcus kohnii ( Staphylococcus cohnii ) FJAT-13685 was cultured at 28 °C for 24 h in the activation medium, the activation medium was tryptone 10 g / L, yeast extract 4 g / L, glucose 2 g / L, magnesium sulfate 1 g / L, phosphoric acid Sodium hydrogen disodium 1 g / L, agar 1.8 g / L, initial pH 6.5;

[0030] 2) Pack 50 mL of culture medium in a 250 mL Erlenmeyer flask, sterilize, cool, and inoculate activated colonies according to conventional methods. After inoculation, culture in a shaker at 30°C and 180 r / min for 18 h; the seed medium is 5 g of glucose / L, yeast extract 10 g / L, tryptone 5 g / L, NaCl 2 g / L, disodium hydrogen phosphate 1.5 g / L, sodium dihydrogen phosphate 1.5 g / L; initial pH 6.5, prepared with deionized water ;

[0031] 3) Fill 50 mL of fermentation medium in a 250 mL Erlenmeyer bottle, sterilize and cool according to conventional methods, and insert the seed fermentation liquid into the fermentation medium according to the inoculum amount of 3%...

Embodiment 3

[0034] 1) with Staphylococcus kohnii ( Staphylococcus cohnii ) FJAT-13685 was cultured at 28°C for 24 h in the activation medium, the activation medium was tryptone 10 g / L, yeast extract 4 g / L, glucose 2 g / L, magnesium sulfate 1 g / L, hydrogen phosphate Disodium 1 g / L, agar 1.8 g / L, initial pH 6.5;

[0035] 2) Pack 50 mL of culture medium in a 250 mL Erlenmeyer flask, sterilize, cool, and inoculate activated colonies according to conventional methods. After inoculation, culture in a shaker at 30°C and 180 r / min for 18 h; the seed medium is 5 g of glucose / L, yeast extract 10 g / L, tryptone 5 g / L, NaCl 2 g / L, disodium hydrogen phosphate 1.5 g / L, sodium dihydrogen phosphate 1.5 g / L; initial pH 6.5, prepared with deionized water ;

[0036] 3) Put 50 mL of fermentation medium in a 250 mL Erlenmeyer flask, sterilize and cool according to conventional methods, and insert the seed fermentation liquid into the fermentation medium according to the inoculation amount of 1%, and inocula...

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Abstract

The invention relates to a staphylococcus cohnii FJAT-13685 and a method for preparing 5-aminolevulinic acid by fermenting the staphylococcus cohnii FJAT-13685, wherein the staphylococcus cohnii FJAT-13685 is preserved with the preservation number of CGMCC No.6323 and the preservation date of July 10, 2012 in the CGMCC (China General Microbiological Culture Collection Center). In the invention, through the adoption of the staphylococcus cohnii FJAT-13685 as a strain and a fermentation medium formed by a carbon source, a nitrogen source, an inorganic salt and a buffer agent, the yield of the 5-aminolevulinic acid can reach 19.8mg / L under an optimized culture condition.

Description

technical field [0001] The invention belongs to the field of biochemical industry, and relates to a strain of staphylococcus kohnii obtained by screening and a method for preparing 5-aminolevulinic acid through culture and fermentation. Background technique [0002] 5-Aminolevulinic acid (5-aminolevulinic acid, ALA) is a precursor substance for the biosynthesis of tetrapyrrole compounds (porphyrin, chlorophyll, heme and vitamin B12) in organisms, and is a key intermediate in the biological regulation of synthesis of tetrapyrrole substances It is widely found in microorganisms, plants and animal cells. [0003] As a photodynamic agent (photodynamic agent, PDT), ALA has a wide range of uses in agrochemicals and medical fields. In the field of agriculture, ALA has multiple functions such as insecticidal, weeding, increasing plant stress resistance and promoting plant growth, and is easy to degrade without residue, non-toxic to humans and animals, and has become a very promisi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P13/00C12R1/44
Inventor 张龙涛朱育菁刘波李昂刘景
Owner FUJIAN ACAD OF AGRI SCI
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