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Method for improving concentration of 1,3-propanediol produced by microbial fermentation

A technology for microbial fermentation and propylene glycol, applied in 1 field, can solve problems such as affecting cell growth and product synthesis, inability of extracellular nutrients to enter into cells, affecting the harvest of fermentation products, etc., and achieves a simple, effective and economical fermentation process, simple control, and ease of use. The effect of industrial application

Inactive Publication Date: 2008-12-17
SOUTHEAST UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The regulation of cell permeability is an important way of microbial metabolic regulation. The absorption of external substances or the secretion of metabolites need to be transported through the cell membrane. In addition, there is a periplasmic space between the cell wall and the cell membrane, which contains proteases, nucleases and transport substances. If there is a barrier between the cell wall and the cell membrane, the intracellular synthetic metabolites cannot be secreted, affecting the harvest of fermentation products, or the extracellular nutrients cannot enter the cell, which also affects cell growth and product synthesis, resulting in a decrease in yield

Method used

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  • Method for improving concentration of 1,3-propanediol produced by microbial fermentation
  • Method for improving concentration of 1,3-propanediol produced by microbial fermentation

Examples

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Effect test

Embodiment 1

[0039] Bacteria: Klebsiella pneumoniae (K.pneumoniae)

[0040] 400ml of fermentation medium containing 45g / L industrial glycerin was placed in a 1L shake flask, and a volume of 40ml of secondary seed solution was inserted, and the temperature was 37°C, and the shaker was cultivated at 180rpm. Add ampicillin respectively at 8 hours of fermentation, so that the content of ampicillin in the fermentation liquid is respectively 0-500 mg / L. Samples were taken at intervals, and the cell concentration was measured by spectrophotometer turbidimetry at a wavelength of 650nm. The cell concentration, that is, the amount of bacteria, was expressed as OD; the concentration of 1,3-propanediol was analyzed by gas chromatography. 24 hours to end fermentation.

[0041] The impact of the amount of ampicillin added on the output of 1,3-propanediol in table 1

[0042]

[0043] Embodiment one has studied in fermenting glycerol production 1,3-propanediol technology, adds ampicillin, concentrati...

Embodiment 2

[0046] 400ml of fermentation medium containing 45g / L industrial glycerol was placed in a 1L shake flask, and a volume of 40ml of secondary seed liquid was inserted, the temperature was 37°C, and the shaker was cultivated at 180rpm. Add 200mg / L ampicillin at 4h, 8h, 12h, 16h and 20h and adjust the pH value. Two parallel samples were averaged, and the experimental results were as follows (Table 2).

[0047] The influence of table 2 ampicillin adding time on 1,3-propanediol output

[0048]

[0049] It can be seen from the above table that the addition of ampicillin at different stages of fermentation has a greater impact on the output of 1,3-propanediol, and the addition of ampicillin in the early and middle stages of fermentation is unfavorable to the biosynthesis of 1,3-propanediol. During this period, the stable growth period has not yet been reached, and the cell wall synthesis of most proliferating cells is incomplete or even the entire cell wall synthesis is blocked, re...

Embodiment 3

[0053] 5L fermenter, liquid volume 3.8L, insert 200ml secondary seed liquid. Control the temperature at 37°C, adjust the pH with 4mol / L NaOH to maintain it at 6.8-7.0, the stirring speed at 300r / min-450r / min, the ventilation rate at 0.4vvm-2.8vvm, and add glycerin to stabilize the concentration at 20g / L- 30g / L, add ampicillin at 12h, 15h, 18h, and 21h respectively, adding 50mg / L each time, and take samples every 4h in the middle to measure the biomass, glycerol concentration and product 1,3-propanediol concentration. After 30 hours of fermentation, the concentration of 1,3-propanediol increased by 43.7% compared with that without adding ampicillin.

[0054] Through the above experiments, it can be proved that in the metabolic pathway of glycerol fermentation to produce 1,3-propanediol, adding antibiotics to increase cell permeability can effectively promote cell growth, and the process of increasing the concentration of 1,3-propanediol, the final product of fermentation, is si...

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Abstract

The invention provides a method for increasing the concentration of 1, 3-propanediol in biological fermentation production, particularly relating to a method for adding an antibiotic, reducing transfer resistance when nutrition and metabolites enter or exit a cell, effectively promoting cell growth and increasing the concentration of the 1, 3-propanediol in the biological fermentation production. The technical process includes the steps that: a secondary seed culture solution is fed into a fermentation culture solution, meanwhile, the antibiotic which can improve cell permeability is added into the fermentation liquor, and the antibiotic is added to change the cell permeability so as to reduce the transfer resistance when the nutrition and the metabolites enter or exit the cell, thus improving the growth capability and the productivity of the thalli and increasing the concentration of the product, 1, 3-propanediol; furthermore, microbes are impelled to excrete the metabolites outside the cells, which decreases the accumulation of the metabolites in the cells, is favorable to eliminate the inhibition of products and by-products, especially reduces the inhibition of the 1, 3-propanediol to cell growth and cell catalytic activity, and at last, the concentration of the fermentation product, 1, 3-propanedio, is improved by 10 percent to 70 percent. The technical process is easy and simple, with low production cost.

Description

technical field [0001] The invention relates to a method for preparing 1,3-propanediol by microbial fermentation, in particular to a method for adding antibiotics, reducing the transfer resistance of nutrients and metabolites into and out of cells, effectively promoting cell growth and increasing the concentration of 1,3-propanediol produced by microbial fermentation , belongs to the technical field of metabolic synthesis of 1,3-propanediol. Background technique [0002] 1,3-propanediol (1,3-PD for short) is a colorless or light yellow viscous liquid at room temperature. 1,3-propanediol reacts with terephthalic acid to polymerize polyester fiber polytrimethylene terephthalate (PTT), which is currently a hot spot in the development of new synthetic fiber varieties. The chemical structure of 1,3-propanediol is HOCH 2 CH 2 CH 2 OH, because it has an odd number of carbons and the difunctional groups are located at both ends, it can be polymerized with other substances to for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/18C12R1/22C12R1/01C12R1/145
Inventor 吴敏苗茂栋郑颖平
Owner SOUTHEAST UNIV
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