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Acid-producing Klebsiella bacterium and uses thereof

A technology of Klebsiella and sodium bromate, applied in the field of bioengineering, can solve the problems of high cost, drop of pH value of fermentation broth, difficult operation, etc., and achieve the effect of improving utilization rate, simple operation and less workload

Inactive Publication Date: 2007-10-31
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cost of the chemical synthesis method of 2,3-butanediol is much higher than the biological method, and compared with the biological method, the chemical method is cumbersome and difficult to operate, so it has been difficult to achieve large-scale industrial production, and its use has no fully developed
The production of these organic acids not only reduces the conversion rate of the substrate, but also causes the pH value of the fermentation liquid to drop during the fermentation process, which is not conducive to the growth of microorganisms, affects the metabolism of bacterial cells, and is not conducive to 2,3-butanediol. biosynthesis of

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Step 1: Treat the strain CCTCC NO: M 207023 with ultraviolet light and diethyl sulfate (DES).

[0042] Take 4 tubes of Klebsiella oxytoca CCTCC NO: M 207023 slant (the slant medium is the LB solid medium) that has grown plumply after 12 hours of cultivation, wash the bacterial lawn with 10 mL of sterile saline, and pour it into a large sterile test tube. Shake the test tube on an oscillating mixer for 30s to break up the bacterial mass, centrifuge at 8000r / min for 15min, discard the supernatant to make a bacterial suspension, count directly under a microscope with a hemocytometer, and adjust the cell concentration to 10 8 pcs / mL, as the bacteria solution to be treated, take 5mL of the bacteria solution to be treated and add it to a sterile petri dish with a diameter of 6cm, put it into a sterile magnetic stirrer, stir and irradiate it under a 15w ultraviolet lamp at a distance of 30cm for 2min Take 5mL of the bacterial suspension and inoculate it into a 250mL Erlenmeyer...

Embodiment 2

[0047] Step 1: Same as Example 1

[0048] Step 2: Fermentation Verification

[0049] Pick a single colony that survived in the selective solid medium, continue to streak culture on the same selective solid medium, after continuous culture for 5 generations and check the genetic stability, pick a single colony and inoculate it in the fermentation medium Carry out fermentation (see general description for fermentation conditions), ferment for 54 hours, get 5mL of fermented liquid 8000r / min centrifugation after 15min, use HPLC to analyze the concentration of 2,3-butanediol and by-product acetic acid and lactic acid in the supernatant, and treatment Compared with the previous results, the results are shown in Table 2. The results showed that under the same fermentation conditions, the 2,3-butanediol yield of strain CCTCC M 207023 was as high as 40.53g / L after treatment, which was 4.86% higher than that before treatment, and the corresponding by-product organic acid lactic acid an...

Embodiment 3

[0052] Step 1: Same as Example 1

[0053] Step 2: Fermentation Verification

[0054]Pick a single colony that survived in the selective solid medium, continue to streak culture on the same selective solid medium, after continuous culture for 5 generations and check the genetic stability, pick a single colony and inoculate it in the fermentation medium Carry out fermentation (see general description for fermentation conditions), ferment for 54 hours, get 5mL of fermented liquid 8000r / min centrifugation after 15min, use HPLC to analyze the concentration of 2,3-butanediol and by-product acetic acid and lactic acid in the supernatant, and treatment Compared with the previous results, the results are shown in Table 2. The results showed that under the same fermentation conditions, the 2,3-butanediol yield of strain CCTCC M 207023 was as high as 40.8g / L after treatment, which was 5.56% higher than that before treatment, and the corresponding by-product organic acid lactic acid and ...

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PUM

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Abstract

The invention discloses a generating acid klebsiella and appliance, which is characterized by the following: setting the preserved number at CCTCC NO: M 207023; adopting ultraviolet and diethyl sulfate; composite-treating strain CCTCC NO: M 207023; seeding on selective solid medium with bromide of sodium and sodium bromate; culturing; sorting single bacterial colony; yeasting; preparing 2, 3-butylene glycol. This invention possesses simple operation, low cost, high efficiency and short period, which can increase receiving ratio of 2, 3-butylene glycol.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a Klebsiella oxytoca, a method for using the Klebsiella oxytoca to biosynthesize 2,3-butanediol and reducing the output of by-product organic acids in the process, And the application of the Klebsiella oxytoca in the biosynthesis process of 2,3-butanediol. Background technique [0002] 2,3-Butanediol is widely used in many fields such as chemical industry, food, fuel and aerospace (Syu M J. Appl Microbiol Biotechnol, 2001, 55(1): 10-18). Its dehydration product, methyl ethyl ketone, is a solvent with a low boiling point, which can be used in coatings, adhesives, lubricants, dyes, inks and other industries, and can also be used as an intermediate for organic synthesis of fragrances and antioxidants; after esterification The dehydration product 1,3-butadiene is an important petrochemical basic organic raw material and synthetic rubber monomer; the octane isomer formed after c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/18C12R1/22
Inventor 黄和纪晓俊李霜杜军练敏高振胡南
Owner NANJING UNIV OF TECH
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