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Gluconobacter oxydans and method for preparing ketoxylose using the same

A technology of Gluconobacter oxydans and xylulose, which is applied in the fields of fermentation engineering and enzyme engineering, can solve the problems such as non-recyclable utilization, unfavorable product separation and purification, and increased production cost.

Active Publication Date: 2009-07-22
SHANDONG TIANLI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is mainly because: on the one hand, cells are in a free state and cannot be recycled; on the other hand, it makes the downstream process very difficult, which is not conducive to the separation and purification of products, and the production cost is greatly increased.

Method used

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  • Gluconobacter oxydans and method for preparing ketoxylose using the same
  • Gluconobacter oxydans and method for preparing ketoxylose using the same
  • Gluconobacter oxydans and method for preparing ketoxylose using the same

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Incline medium: glucose 30g / L, yeast extract 10g / L, beef extract 5g / L, agar 20g / L.

[0045] Shake flask medium: glucose 30g / L, yeast extract 25g / L, beef extract 5g / L, D-arabitol 10g / L, KH 2 PO 4 5g / L.

[0046] Cultivate Staphylococcus oxidans CGMCC No.2709 on a slant medium at 30°C for 24 hours, then put a ring of this bacteria in the shake flask medium, cultivate it at 30°C for 10 hours, and the shake flask rotates at a speed of 200r / min. The content of D-ara is 20g / L, and the enzyme production is 2.5~5.0U / mL. It is transformed by free cells. Weigh 1g of wet cells after centrifugation and add them to 10mL of 30g / L D-ara solution (the amount of cells added is calculated as per 100g of cells transformed 1L of 30g / L D-ara solution), under the condition of 30°C, stir and oscillate in a shaker flask (rotating speed 120r / min) to convert and produce D-xylulose. The reaction time is 9h. After the reaction is terminated, the substrate conversion rate and product concentratio...

Embodiment 2

[0048] With 30g / L maltose as the carbon source, 20g / L yeast extract, and 10g / L bean cake powder as the nitrogen source, the other components of the medium and the cell culture conditions are the same as in Example 1, and a 1.5% sodium alginate colloid solution is prepared, and the physiological After mixing the bacterial suspension prepared with saline evenly, add 8% diatomaceous earth for adsorption, and drop 2% CaCl with a syringe 2 Solidify in the solution to make spherical immobilized cells with a diameter of about 3-4mm, and put them in a refrigerator at 4°C for several hours, then pour off the supernatant, wash twice with distilled water, and replace 1g of the centrifuged wet cells with seaweed 5 g of immobilized cells were obtained by embedding in sodium phosphate, and transformed into 10 mL of 50 g / L D-arabitol solution in a shake flask at 30°C (rotating speed: 200 r / min). The transformation time for each batch was 13 hours, and 30 batches were transformed. , the avera...

Embodiment 3

[0050] Glucose 225g, yeast extract 112.5g, beef extract 22.5g, D-arabitol 45g, KH 2 PO 4 22.5g, pH 5.0, dilute to 4.5L with water, make culture medium, put into 7.5L stirring fermenter, steam sterilize at 121℃ for 15min. Prepare seed medium: glucose 30g / L, yeast extract 10g / L, beef extract 5g / L. Put Gluconobacter oxydans CGMCCNo.2709 in the seed medium one by one, and culture it at 30°C for 8 hours to obtain the seed liquid. Put the seed liquid into the cooled fermentation medium according to the inoculation amount of 3% of the volume of the fermentation liquid, and cultivate it at 30°C. (Ventilation 0.7vvm, stirring speed 600r / min) 48h. The fermented liquid is subjected to bacterial filtration treatment in an ultrafilter, and the molecular weight cut-off of the ultrafiltration membrane is 10 6 Da, the operating pressure is 0.2MPa, the temperature is controlled at 50°C, and the membrane surface velocity is 4m / s. After immobilizing the 100g bacterium obtained by interceptio...

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Abstract

The invention discloses a Gluconobacter oxydans, which is classified and named as Gluconobacter oxydans NH-10, and preserved in China General Microbiological Culture Center of Microbial Culture Collection Management Committee with the preservation number of CGMCC No.2709. The invention further discloses a D-xylulose preparation method that uses the Gluconobacter oxydans. The strain obtained by screening can transform D-Arabitol so as to generate D-xylulose in high efficiency, the D-xylulose preparation method reaches the highest D-Arabitol transformation ratio of 99.5 percent (w / w) and the D-xylulose yield ratio of 95 percent, the D-xylulose concentration of a conversion fluid can reach 95g / L, and the conversion fluid hardly contains other ingredients.

Description

technical field [0001] The invention belongs to the technical fields of fermentation engineering and enzyme engineering, and relates to a D-arabitol dehydrogenase-producing microorganism Gluconobacter oxydans and a method for using it to produce D-xylulose. Background technique [0002] Xylulose (xylulose) is a ketopentose. L-xylulose is excreted in the urine of pentoseuria. In vivo, the L-type and D-type xylulose are first reduced to xylitol, and then reoxidized to convert each other. D-xylulose reacts with ATP to generate D-xylulose-5-phosphate and enters the pentose phosphate pathway. In addition, glucose forms D-xylulose-5-phosphate through 6-phosphogluconic acid→D-ribulose-5-phosphate, which is an important intermediate in the pentose phosphate pathway of another glycolysis pathway. In addition, D-xylulose can be used as a material for the production of xylitol, which is one of the important sweeteners in the food industry and other fields. Xylitol is a natural five...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/02C12R1/01
Inventor 徐虹朱宏阳李莎赵敏蔡恒
Owner SHANDONG TIANLI PHARMA
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