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Microbial cell conversion method for producing 2-keto-D-gluconic acid

A microbial cell and gluconic acid technology, which is applied in the field of microbial cell transformation to produce 2-keto-D-gluconic acid, can solve the problems of large waste residue treatment, large heat release, and affecting enzyme activity, and achieve simple separation and purification. The effect of saving production cost and high conversion rate

Active Publication Date: 2014-03-12
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a method for the production of 2-keto-D-gluconic acid by microbial cell transformation, thereby solving the long reaction time and low yield of the chemical synthesis method of 2-keto-D-gluconic acid in the prior art , The reaction conditions are harsh, the production cost of the enzymatic method is high, the generation of hydrogen peroxide affects the enzyme activity, and the fermentation method has the defects of long fermentation period, large heat release, high extraction cost, and large amount of waste residue treatment

Method used

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  • Microbial cell conversion method for producing 2-keto-D-gluconic acid
  • Microbial cell conversion method for producing 2-keto-D-gluconic acid

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Effect test

Embodiment 1

[0041] Transfer the aqueous solution containing 200g / L glucose into a 7L fermenter, add 60g / L resting cells, and react under the conditions of 30°C, air ventilation rate 8L / min, and stirring speed 600rpm, and adjust with 2M NaOH solution during the reaction process. The pH was maintained at 6.0, and the glucose in the supernatant was completely converted in about 21 hours. The yield of 2-KGA (sodium salt) was 97.75%, the concentration could reach 234.6g / L, and the space-time yield was 11.17g / L / h.

Embodiment 2

[0043] Transfer the aqueous solution containing 250g / L glucose into a 7L fermenter, add 60g / L resting cells, and react under the conditions of 30°C, air flow rate 8L / min, stirring speed 600rpm, and adjust with 2M NaOH solution during the reaction process. The pH was maintained at 6.0, and the glucose in the supernatant was completely converted in about 34 hours. The yield of 2-KGA (sodium salt) was 94.74%, the concentration could reach 284.22g / L, and the space-time yield was 8.36g / L / h.

Embodiment 3

[0045] Transfer to an aqueous solution containing 200g / L sodium gluconate in a 7L fermenter, add 30g / L resting cells, and react under the conditions of 30°C, air ventilation 8L / min, and stirring speed 600rpm, and in the supernatant about 15h The conversion of gluconic acid is complete, the concentration of 2-KGA (sodium salt) can reach 198.2g / L, and the space-time yield is 13.2g / L / h.

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Abstract

The invention provides a microbial cell conversion method for producing 2-keto-D-gluconic acid. The method comprises the steps of A. preparing resting cells with a genetic engineering strain of gluconobater oxydans DSM2003 as a strain, wherein the genetic engineering strain is gluconobater oxydans of overexpressing gluconic acid dehydrogenase, and the gene of the gluconic acid dehydrogenase comes from the gluconobater oxydans and has a nucleotide sequence as shown by SEQ ID NO:1; B. preparing a conversion liquid containing glucose or gluconic acid, wherein the concentration of the glucose is 200-250g / L, and the concentration of the gluconic acid is 200-300g / L; and C. adding 30-60g / L of resting cells into the conversion liquid for converting so as to obtain the 2-keto-D-gluconic acid. The method has the advantages that the conversion rate is high, the product is single, the separation and purification are simple, the production cost is saved, the cells are easy to recycle and are reusable, and the like.

Description

technical field [0001] The invention relates to the field of microbial cell transformation, in particular to a method for producing 2-keto-D-gluconic acid through microbial cell transformation. Background technique [0002] 2-keto-D-gluconic acid (2-keto-D-gluconic acid, 2-KGA) is one of the organic acids produced on a large scale in the world, mainly used as D-isoascorbic acid (EA) and its salts (such as D-sodium erythorbate, EN), and EA and EN, as isomers of vitamin C, have strong antioxidant properties. As a metabolizable antioxidant, their antiseptic effect greatly exceeds that of vitamin C. Vitamin C, and the price is less than half of vitamin C, the application prospect is very extensive. At present, EN has been widely used as a new type of food antioxidant, antiseptic and fresh-keeping agent in meat, fruit, beverage and other foods. 2-KGA also has a variety of industrial uses, such as feed additives, detergent accelerators, cement plasticizers and photographic devel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/58C12N15/74C12N1/21C12R1/01
Inventor 林金萍魏东芝李克非
Owner EAST CHINA UNIV OF SCI & TECH
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