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A method for increasing the biomass of Gluconobacter futau hd924

A technology of HD924 and Fotoglucose, applied in the field of bioengineering, can solve the problem of low biomass

Active Publication Date: 2019-09-03
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to overcome the defect of low biomass of Gluconobacter HD924 in the prior art in the process of fermenting dihydroxyacetone, and provide a method for increasing the biomass of Gluconobacter HD924

Method used

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  • A method for increasing the biomass of Gluconobacter futau hd924
  • A method for increasing the biomass of Gluconobacter futau hd924
  • A method for increasing the biomass of Gluconobacter futau hd924

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Experimental program
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Embodiment

[0012] In this example, G. frateurii HD924 was used as the experimental strain, and Bacillus lentus 168 was purchased from the American Type Culture Collection.

[0013] Determination of DHA by HPLC method: take 1 mL of fermentation broth, centrifuge at 12000 r / min for 10 min, take 0.5 mL of supernatant and dilute with distilled water to the linear range of determination (0-10 g / L), membrane filter, and then enter HPLC for determination. The DHA content in the fermentation broth was calculated by external standard method. Chromatographic separation conditions are: Aminex HPX-87H chromatographic column (300mm×7.8mm, 9μm), UV detector detection wavelength is 260nm, mobile phase: 8mmol / L sulfuric acid. The column temperature was 50 °C, the flow rate was 0.5 mL / min, and the injection volume was 30 μL.

[0014] Glycerin detection method: The glycerin content was determined by HPLC. The determination conditions are the same as DHA, and the detector is RI detector.

[0015] Coenzy...

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Abstract

The invention discloses a method for increasing the biomass of gluconobacter frateurii HD924, and belongs to the field of bioengineering. By overexpressing a glucose dehydrogenase (gdh) gene from Bacillus lentus 168 through a constitutive promoter sldAp in the gluconobacter frateurii HD924 and meanwhile adding L- glucose with the certain concentration into a culture medium, the dry cell weight ofrecombinant bacteria is increased by 21.2%. According to the recombinant gluconobacter frateurii overexpressing the gdh gene, the yield of dihydroxyacetone is remarkably increased, the production intensity is remarkably improved, the glycerinum utilization rate is remarkably increased, and the intracellular NADH concentration is remarkably increased.

Description

technical field [0001] The invention relates to a method for increasing the biomass of gluconobacter futau HD924, which belongs to the field of bioengineering. Background technique [0002] Gluconobacter frateurii (Gluconobacter frateurii) HD924 is a Gram-negative bacteria, obligate aerobic, belonging to the genus Gluconobacter. The preservation number of the bacterial strain in the General Microorganism Center of China Microbiological Culture Collection Management Committee is: CGMCC No.5397. The bacterial strain has excellent dihydroxyacetone production performance, and the dihydroxyacetone production can reach more than 170 g / L under aerobic conditions (CN, 201110388519). The strain has been industrially produced in Changxing Pharmaceutical Co., Ltd., and is one of the industrial strains producing dihydroxyacetone. Liu Yupeng et al. (Bioresource technology 2013, 142:384-389) found that the maximum biomass OD560 in the fermentation process of Gluconobacter fortoria HD924...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P7/26C12R1/01
Inventor 李华刘宇鹏肖延铭杨生玉杨卫华
Owner HENAN UNIVERSITY