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Method for synthesizing 1,3-dioxyacetone by glycerol fermentation

A technology of dihydroxyacetone and glycerol, applied in 1 field, can solve the problems such as difficulty in improving the yield of DHA, lysing and inactivation of fermentation cells

Inactive Publication Date: 2011-11-23
XUZHOU HENGYUAN BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the main problem of this method is that when the concentration of the substrate glycerol and the product DHA in the medium is too high, high osmotic pressure is generated, which makes the fermentation cells lyse and inactivate, thus making it difficult to increase the yield of DHA.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Bacillus licheniformis isolation medium : Distilled water 1000mL, glycerin 5g, NaCl 5g, yeast powder 5g, glucose 20g, agar powder 15g, adjust pH to 7.0, sterilize at 0.1MPa for 20min, cool to 35℃~45℃, add Nile Red 2mL / L (0.30mg Nile Red dissolved in 100mL dimethyl sulfoxide), poured into a Petri dish under sterile conditions, cooled and set aside.

[0016] Nutrient rich medium : Distilled water 1000mL, yeast powder 10g, agar powder 10g, glycerin 3g, (NH 4 ) 2 SO 4 5g, adjust the pH to 7.0, and sterilize at 0.1MPa for 10min.

[0017] Product fermentation medium:

[0018] Preparation of Phosphate Buffer : Distilled water 1000mL, NaH 2 PO 4 .12H 2 O 8.95g, KH 2 PO 4 1.5g, sterilized at pH7.00.1MPa, 15min-20min, for later use.

[0019] Enrichment medium for genetically engineered bacteria is LB medium : The fermentation medium is the LB medium that adds 15.0% glycerol, and air is blown in as the oxidant of the genetically engineered bacteria; the culture te...

Embodiment 2

[0024] The composition and culture conditions of each medium are the same as above.

[0025] Glycerin fermentation : Pre-culture is in the L-test tube, add 5ml nutrient-enriched medium with aseptic operation, and insert a single colony of Bacillus licheniformis B-05571 with a sterile toothpick. Cultivate at 33°C and 120r / min for 20h. Inoculate 0.5 ml of the pre-culture into a 500 ml Erlenmeyer flask containing 100 ml of nutrient-enriched medium, and cultivate for 30 h at 30°C with shaking at 120 r / min. Sterile centrifuge at 6000*g for 10min at 4°C, discard the supernatant, shake and mix with sterile phosphate buffer in a centrifuge tube, then centrifuge at 6000*g for 12min at 4°C, discard the supernatant. Inoculate 0.5 ml of the pre-culture into a 500 ml Erlenmeyer flask containing 100 ml of nutrient-enriched medium, and incubate at 35°C for 24 hours with shaking at 130 r / min. Sterile centrifuge at 6000*g for 10min at 4°C, discard the supernatant, vortex and mix with steri...

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Abstract

The invention relates to the synthesis of a 1,3-dioxyacetone. A method for synthesizing 1,3-dioxyacetone by glycerol fermentation comprises the following steps: performing the gene recombination and DNA rearrangement technology on bacillus licheniformis B-05571 separated and screened from acid soil, integrating a dehydrogenase gene into an Escherichia coli JM109, constructing industrial engineering bacteria of 1,3-dioxyacetone synthesized by glycerol dehydrogenization, immobilizing by using a supermolecule self-assembly template, then fermenting in a culture medium containing glycerol to synthesize 1,3-dioxyacetone. The product 1,3-dioxyacetone can be obtained by the steps of filtering fermentation liquor, extracting with an organic solvent and recrystallizing with a combined solvent. The synthesis process route is simple and convenient, the product is high in quality and low in cost, and the total mass yield is higher than 75%.

Description

[0001] Technical field: the invention belongs to the technical field of bio-organic synthesis, and relates to a bio-organic synthesis method of 1,3-dihydroxyacetone. Background technique [0002] 1,3-Dihydroxyacetone is an important chemical and biochemical raw material, a synthetic intermediate of medicine and pesticide, and a multifunctional food additive, with a wide range of uses. The synthesis methods of dihydroxyacetone mainly include precious metal catalytic oxidation method and microbial method. The synthesis method of heavy metal catalyzed oxidation of glycerol, because the fatal shortcoming of poor catalytic selectivity cannot be overcome, causes the conversion rate of glycerol to be lower than 40%, the productive rate of DHA is lower than 25%. The industrial production method currently utilizes microbial batch fermentation. This method is to use the dehydrogenase produced by the bacteria to carry out dehydrogenation reaction with glycerol as the substrate when the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/26C12N1/21C12N15/53C12R1/19
Inventor 傅尧郭庆祥曹文化封立刚腾道路邓晋赵红李玉玲徐清张颖
Owner XUZHOU HENGYUAN BIOENG
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