Method using klebsiella pneumoniae to produce xylonic acid

A technology of pneumonia bacillus and xylonic acid, which is applied in the biological field to achieve the effects of high production intensity, high final product concentration and high conversion rate

Active Publication Date: 2017-05-24
SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Method using klebsiella pneumoniae to produce xylonic acid
  • Method using klebsiella pneumoniae to produce xylonic acid
  • Method using klebsiella pneumoniae to produce xylonic acid

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Embodiment 1

[0025] The gene recombination method is used to inactivate the Klebsiella pneumoniae gluconate dehydrogenase gene to realize the inactivation of the glucose dehydrogenase activity.

[0026] Klebsiella pneumoniae in this embodiment adopts the bacterial strain of numbering CGMCC 1.6366 (this bacterial strain is also referred to as TUAC01 or ACO1), and the bacterial strain of CGMCC 1.6366 has been disclosed in the authorized invention patent ZL201210093993, and also in the open literature (Wei Dong, Wang This strain has also been disclosed in Min, Shi Jiping, Hao Jian. Red recombinase assisted gene replacement in Klebsiellapneumoniae. Journal of Industrial Microbiology & Biotechnology. 2012 39:1219-1226). This strain is a strain used to produce 1,3-propanediol, 2,3-butanediol, acetoin and 2-ketogluconic acid. The bacterium was isolated from soil, and the isolation process and traits are described in open literature (Hao Jian, et al. Isolation and characterization of microorganism...

Embodiment 2

[0044] The Klebsiella pneumoniae bacterial strain Kp-gad that the gluconate dehydrogenase activity obtained in wild-type bacterial strain Klebsiella pneumoniae and embodiment 1 is eliminated - Shake flask batch fermentation experiments were performed.

[0045] The Klebsiella pneumoniae bacterial strain Kp-gad that the gluconate dehydrogenase activity obtained in the wild-type bacterial strain Klebsiella pneumoniae CGMCC 1.6366 and embodiment 1 is eliminated - They were respectively inoculated into 250ml Erlenmeyer flasks containing 50ml xylose acid fermentation medium, the rotation speed of the shaker cabinet was 200 rpm, and the aerobic fermentation was carried out at a constant temperature of 25°C.

[0046] The components of the fermentation medium are: xylose 5g / L, corn steep liquor 0.5g / L, calcium carbonate 1g per bottle.

[0047] After culturing for 24 hours, the components in the fermentation broth were determined. It is determined by liquid chromatography, and the com...

Embodiment 3

[0052] The Klebsiella pneumoniae bacterial strain Kp-gad that the gluconate dehydrogenase activity obtained in wild-type bacterial strain Klebsiella pneumoniae and embodiment 1 is eliminated - Performing shake flask batch fermentation experiments

[0053] Kp-gad obtained in the wild-type bacterial strain Klebsiella pneumoniae CGMCC 1.6366 and embodiment 1 - Inoculate into 250ml Erlenmeyer flask respectively, wherein the fermentation medium of the mixture of 50ml glucose and xylose is housed, shake the bottle cabinet rotating speed 200 revs per minute, constant temperature 45 ℃ carry out aerobic fermentation culture.

[0054] The medium components are: glucose 10g / L, xylose 5g / L, yeast extract 0.5g / L, calcium carbonate 1g per bottle.

[0055] After culturing for 24 hours, the components in the fermentation broth were determined. The product was detected by the method in Example 2. Gluconic acid and gluconate are not distinguished in the present invention, and xylose and xylo...

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Abstract

The invention discloses a method using klebsiella pneumoniae to produce xylonic acid. The method comprises the following steps of using xylose as raw material, and enabling the klebsiella pneumoniae to convert the xylose into xylonic acid under the aerobiotic condition; furthermore, utilizing the klebsiella pneumoniae with deactivation of gluconate dehydrogenase to convert the xylose into the xylonic acid, so as to improve the conversion rate from xylose into xylonic acid; furthermore, utilizing a two-step fermenting method, enabling a fermenting liquid to maintain the neutral pH (potential of hydrogen) condition in the first fermenting phase, enabling the fermenting liquid to maintain the acid fermenting condition in the second fermenting phase, and fermenting to produce the xylonic acid. The method has the advantages that the primer conversion rate is higher, the production intensity is higher, and the product final concentration is higher; by using the glucose and xylose mixture as the raw material, the gluconic acid and the xylonic acid can be simultaneously produced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for producing xylonic acid by Klebsiella pneumoniae. Background technique [0002] Xylose is the main constituent monomer of hemicellulose, and the utilization of xylose has been a hot spot that people have paid attention to in recent decades. Xylose acid is a derivative of xylose. Xylose acid is similar in structure and properties to gluconic acid. It can replace gluconic acid in many applications. Xylonic acid is the raw material for the production of 1,2,4-butanetriol and copolyamides. In addition, xylonic acid can also be used as a concrete additive, and its water-reducing effect is better than gluconic acid. [0003] The production of xylonic acid is mostly produced by chemical or biosynthetic methods using xylose as raw material. The chemical method to generate xylonic acid is mainly oxidized by a catalyst. For example, in a strong alkaline environment, ...

Claims

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Application Information

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IPC IPC(8): C12P7/58C12R1/22
Inventor 郝健王晨红王德信史吉平姜标
Owner SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
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