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Method for 3-hydroxyl ethylformic acid fermentation production

A technology of hydroxypropionic acid and sodium propionate, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., to achieve the effects of short growth cycle, simple medium and low cost

Inactive Publication Date: 2009-06-17
青岛生物能源与过程研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the patent, Cargill designed 3 pathways to produce 3-hydroxypropionate from glucose through pyruvate fermentation, including lactic acid pathway, acetyl-CoA pathway and β-alanine pathway, and laboratory experiments proved the feasibility of these processes , but the stability of the target gene in the cell, the expression level and activity of the enzyme, the tolerance of the host cell to the

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The following are percentages by weight.

[0047] Dissolve yeast powder 0.5%, peptone 1.2%, sodium propionate 0.4%, potassium dihydrogen phosphate 0.05%, magnesium sulfate 0.05% in 750ml of water, adjust pH to 5.70 with 5mol / L NaOH solution or 3mol / L hydrochloric acid solution Afterwards, sterilize.

[0048] Take by weighing 0.003% ferric sulfate heptahydrate of the total amount of the culture medium and add 40ml of water to dissolve it, add 4ml of hydrochloric acid with a concentration of 1 / 5 (v / v) to dissolve it, make the solution clearer, filter it with a sterile filter membrane, and dissolve it with sterilized The above solutions of bacteria were mixed under aseptic conditions.

[0049] Mix the sterilized sucrose with the above-mentioned cooled medium to obtain 5% sucrose, namely to obtain the fermentation medium.

Embodiment 2

[0051] (1) Seed culture of Candida rugosa

[0052] The cultivation time is 20 hours, the cultivation temperature is 30°C, and the components and weight contents of the seed medium are:

[0053] Glucose 2%, yeast powder 0.4%, peptone 1.5%, dipotassium hydrogen phosphate 0.06%, magnesium sulfate 0.05%, iron sulfate 0.002%. Adjust the pH to 5.5 with NaOH or HCl solution.

[0054] (2) Fermentation culture

[0055] The volume of the initial fermentation medium is 1.8L, and the composition of the initial medium used is as follows:

[0056] Sucrose 5%, yeast powder 0.5%, peptone 1.2%, sodium propionate 0.4%, potassium dihydrogen phosphate 0.05%, magnesium sulfate 0.05%, iron sulfate 0.002%. Adjust the pH to 5.5 with NaOH or HCl solution.

[0057] The concentration of each component of the feeding solution is:

[0058] Feed 1: 40wt% sucrose aqueous solution;

[0059] Feed 2: industrial amino acid powder aqueous solution 10wt% (adding in the second stage)

[0060] Inoculate the ...

Embodiment 3

[0065] 3-Hydroxypropionic Acid Production by Candida rugosa in Raw Medium

[0066] Medium components: glucose 5%, yeast powder 0.4%, peptone 1.2%, dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.05%, iron sulfate 0.005%. Adjust the pH to 5.5 before sterilization.

[0067] Insert the Candida rugosa seed solution with an inoculum of 5% by volume, cultivate it on a shaker at 30°C at 300rpm, add NaOH and sulfuric acid to control the pH value of the culture solution to 5.3±2.0, and the sugar is consumed in 50 hours, measuring 3 -Hydroxypropionic acid yield is 6g L -1 .

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Abstract

The invention discloses a method for producing 3-hydracrylic acid by fermentation comprising inoculating candida rugosa seeds in a culture medium, stirring and controlling the pH value to 5.3+ / -0.2; adding a outcome promoting factor, a sugar aqueous solution and a amino acid aqueous solution with a pH of 4.7+ / -0.2; finally adding another sugar aqueous solution with a pH of 4.2+ / -0.2, fermenting and culturing to obtain the fermentation solution containing 3-hydracrylic acid. The method of the invention employs a static optimization method combining a single factorial experiment, an orthogonal experiment and response surface experiment, optimizes the culture medium and the 3-hydracrylic acid production by pH sectional regulation, enables the yield of the 3-hydracrylic acid to be greatly improved to more than 28g / L, the conversion of the sugar to be 0.32g / g, the production ability to be 0.54g / Lh, and has an industrial popularization and application prospect.

Description

technical field [0001] The invention relates to a method for producing 3-hydroxypropionic acid, in particular to a method for fermenting and producing 3-hydroxypropionic acid by adopting Candida rugosa. Background technique [0002] 3-Hydroxypropionic acid (3-HP for short, the same below) is a three-carbon achiral organic acid, which is isomers with lactic acid and has two functional groups, hydroxyl and carboxyl. 3-Hydroxypropionic acid is colorless, odorless, oily, miscible with various organic solvents such as water, alcohol, ether, etc., and is widely used in the production of coatings, adhesives, water treatment chemicals and personal care products; at the same time, 3-hydroxypropionic acid Acid is an important chemical intermediate emerging in recent years. It is dehydrated to generate acrylic acid, oxidized to generate malonic acid, esterified with alcohol to generate ester, and reduced to generate 1,3-propanediol. [0003] At present, the preparation method of 3-hyd...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/42C12R1/72
Inventor 朱建航辜旭辉刘寅
Owner 青岛生物能源与过程研究所
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