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Rapid screening method for dihydroxypropanone high-yield fungus

A high-yield technology for dihydroxyacetone and hydroxyacetone, applied in biochemical equipment and methods, microbe measurement/inspection, fermentation, etc., can solve problems such as blanks, and achieve the effects of reducing labor, improving labor efficiency, and simple operation

Inactive Publication Date: 2008-07-23
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although there are many microorganisms that convert glycerol into dihydroxyacetone, there are relatively few high-yielding bacteria that are actually used to produce DHA, and the reports on the rapid screening of microorganisms that convert glycerol into dihydroxyacetone are blank.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) Preparation of Fehling's reagent:

[0038] A liquid with 34.6gCuSO 4 ·5H 2 O was dissolved in 200 mL of water, and 0.5 mL of concentrated H 2 SO 4 , after mixing, dilute to 500mL with water,

[0039] B liquid uses 173g potassium sodium tartrate (KNaC 4 h 4 o 6 4H 2 (2) and 71g NaOH solid are dissolved in 400mL water, then diluted into 500mL solution,

[0040] When in use, take equal amounts of liquid A and liquid B and mix them for immediate use;

[0041] (2) Preparation of medium:

[0042] Plate medium, multi-well plate medium and slant medium preparation: the plate medium, multi-well plate medium and slant medium are solid medium with the same composition, and its components are as follows: glycerol 20g, yeast powder 2g, Agar 18g, water 1000mL, pH value 5.0. It is prepared as follows: add 18g of agar to 1000mL of water under heating conditions to dissolve it, then add 20g of glycerin and 2g of yeast powder, sterilize at 121°C for 20min, and when cooled ...

Embodiment 2

[0054] (1) Preparation of Fehling's reagent:

[0055] Liquid A: use 34.6g CuSO 4 ·5H 2 O was dissolved in 200 mL of water, and 0.5 mL of concentrated H 2 SO 4 , after mixing, dilute to 500mL with water;

[0056] Solution B: use 173g potassium sodium tartrate (KNaC 4 h 4 o 6 4H 2 (2) and 71g NaOH solid are dissolved in 400mL water, then diluted into 500mL solution;

[0057] When in use, take equal amounts of liquid A and liquid B and mix them for immediate use;

[0058] (2) Preparation of medium:

[0059] Plate medium, multi-well plate medium and slant medium preparation: the plate medium, multi-well plate medium and slant medium are solid medium with the same composition, and its components are as follows: glycerol 30g, yeast powder 3g, Agar 20g, water 1000mL, pH value 6.0; prepare as follows: Add 20g agar to 1000mL water under heating condition to dissolve it, then add 30g glycerin and 3g yeast powder, sterilize at 121°C for 20min, cool to 60°C, In a sterile enviro...

Embodiment 3

[0070] (1) Preparation of Fehling's reagent:

[0071] A liquid with 34.6g CuSO 4 ·5H 2 O was dissolved in 200 mL of water, and 0.5 mL of concentrated H 2 SO 4 , after mixing, dilute to 500mL with water,

[0072] B liquid uses 173g potassium sodium tartrate (KNaC 4 h 4 o 6 4H 2 (2) and 71g NaOH solid are dissolved in 400mL water, then diluted into 500mL solution,

[0073]When in use, take equal amounts of liquid A and liquid B and mix them for immediate use;

[0074] (2) Preparation of medium:

[0075] Plate medium, multi-well plate medium and slant medium preparation: the plate medium, multi-well plate medium and slant medium are solid medium with the same composition, and its components are as follows: glycerin 50, yeast powder 5, Agar 20, water 1000mL, pH value 7.0, prepared as follows: Add 20g of agar to 1000mL of water under heating conditions to dissolve it, then add 50g of glycerin and 5g of yeast powder, sterilize at 121°C for 20min, cool to 60°C,

[0076] In...

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Abstract

The invention relates to a fast screening method for dihydroxyacetone high-yielding strain, which is characterized in that a waiting colony is inoculated in the culture medium of perforated distribution plate until a single colony is grown; the fehling reagent is added to test; if a brown-red halo spot is shown around the single colony, the single colony is a positive strain; the positive strain is fetched to be inoculate to the inclined culture medium, and cultivated at 28 to 37 DEG C in 1 to 2 days; the inclined strain of the positive strain is obtained; the inclined strain is inoculated in the fermentation medium, and the zymotic fluid is cultivated by oscillation; the content of DNA in the zymotic liquid is determined; the DNA content of the zymotic fluid obtained by fermentation is higher than the DNA content of the 2 g / L positive strain, which is dihydroxyacetone high-producing strain. The fast screening method for dihydroxyacetone high-yielding strain has the advantages of simple operation, visual and convenient judgment to result, fast and high-flux screening of microbial strain well producing dihydroxyacetone, significantly decreased amount of labor, and increased labor efficiency.

Description

(1) Technical field [0001] The invention relates to a rapid screening method for dihydroxyacetone high-yielding bacteria, which belongs to the field of industrial microbe engineering. (2) Background technology [0002] Dihydroxyacetone, also known as 1,3-dihydroxyacetone (1,3-Dihydroxyacetone) (hereinafter referred to as DHA), is the simplest ketose, white powdery crystal with sweet taste, easily soluble in water and ethanol , acetone and ether and other organic solvents. This chemical has a molecular weight of 90.08. [0003] DHA has a wide range of uses and a large market capacity. The research on the production of dihydroxyacetone by biotransformation of glycerol is of great significance. Compared with chemical synthesis, the production of DHA by microbial transformation has many advantages: strong specificity, mild reaction conditions, and substrate utilization. High, high conversion rate, simple production process and other advantages. [0004] There are many patents...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12P7/26
Inventor 郑裕国胡忠策薛亚平沈寅初
Owner ZHEJIANG UNIV OF TECH
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