Method for fermentation preparation of Ectoine

A technology of microbial fermentation and concentration, applied in the field of microbial fermentation, can solve the problems of high price of transcription inducer, low synthesis amount, and no obvious improvement of Ectoine fermentation yield.

Active Publication Date: 2008-12-03
DALIAN MARITIME UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this study solved the problem of Ectoine secretion, the fermentation yield of Ectoine was different from that of non-secreting strains because of the slow growth of cells, long culture time, reaching the highest OD value of growth in about 90 hours, and the low total synthesis of Ectoine. Compared with "bacterial milking technology", it has not been significantly improved
In addition, the synthetic and secreted Ectoine genetic engineering Escherichia coli constructed by Schubert et al., the Ectoine fermentation yield was 0.8g / L / d (Continuous synthesis and excretion of compatible solute ectoine by a transgenic, nonhalophilic bacterium., Appl.Environ.Microbiol. , 73, 3343-3347, 2007), except that the fermentation yield of Ectoine is low, the price of the inducer for initiating transcription in the medium of genetically engineered E. coli is expensive
[0005] The common shortcoming of the Ectoine fermentation method in the prior art is that cell growth is slow, and Ectoine synthesis amount is low
Through high-density fermentation, although higher cell densities can be achieved, fermentation yields cannot be significantly increased due to slow cell growth and low synthesis

Method used

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  • Method for fermentation preparation of Ectoine
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  • Method for fermentation preparation of Ectoine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Strain: H. salina DSM 5928;

[0034] Medium: L-monosodium glutamate 30g / L, KH 2 PO 4 3g / L, K 2 HPO 4 9g / L, MgSO 4 ·7H 2 O0.4g / L, MnSO 4 4H 2 O 0.01g / L, NaCl 60g / L, pH7.2, 0.2μm filter sterilization;

[0035] Cultivation method: After the bacterial strain was cultured in a test tube containing 5 mL of the above-mentioned medium at 30°C and on a shaker (120rpm) for 24 hours, it was transferred to 30mL of the above-mentioned medium (300mL Erlenmeyer flask) by 1% (volume) inoculation amount ), cultured at 30°C with a shaker at 120rpm, and sampled for 36 hours to measure the concentration of Ectoine inside and outside the cells;

[0036] Results: According to molecular weight and structure identification and analysis, there was secreted Ectoine in the culture medium; the concentration of Ectoine in cells was 0.56g / L, the concentration of Ectoine in culture medium was 0.68g / L, and the secretion rate was 55%.

Embodiment 2

[0038] Bacterial strains: Halomonas (Halomonas) and several other microorganisms, see Table 1;

[0039] Culture medium condition and culture method are as embodiment 1; Experimental result is shown in table 1:

[0040] Table 1

[0041]

[0042] (to be continued)

[0043] (Continued from Table 1)

[0044]

[0045] It can be seen from the results in Table 1 that under the same culture conditions, not all Ectoine synthetic strains can secrete Ectoine.

Embodiment 3

[0047] This example examines the effect of adding yeast extract on the synthesis and secretion of Ectoine by H. salina DSM 5928.

[0048] In this embodiment, on the basis of Example 1, 0 to 10 g / L of yeast extract is added to the medium, and Ectoine is induced and synthesized according to the culture conditions of Example 1, and the total amount of Ectoine synthesized (intracellular Ectoine amount and secretion The sum of Ectoine amount) and secretion amount, the results are shown in the appendix figure 1 shown.

[0049] It can be seen that the total synthesis and secretion of Ectoine decreased with the increase of yeast extract concentration, especially when the yeast extract concentration reached 10g / L, H.salina DSM 5928 only secreted a small amount of Ectoine.

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Abstract

The invention relates to a method for fermenting and generating Ectoine by utilizing microorganisms, which belongs to the microorganism fermentation engineering filed. The method comprises the following steps: fermenting a plurality of strains of Halomonas, which can secrete Ectoine, in a liquid culture medium with glutamic acid mono sodium as a sole carbon nitrogen source and the NaCl concentration being 15 to 120g/L; inducing synthesis and enaling Ectoine to be secreted; and recycling the Ectoine in a fermentation system. The method ensures that the fermentation productivity of the Ectoin reaches 6.4 to 7.9g/L/d, which is 2.1 to 2.4 times of that of the prior art, and products are mostly secreted into the culture medium, so as to simplify the recycling process of the products, and to be favor of realizing continuous culture. Compared with the prior art, the NaCl concentration in the culture medium is reduced by 1/2 to 2/3, thus being favor of reducing the corrosion of equipment, and reducing the downstream processing cost.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation and relates to a method for producing Ectoine by microbial fermentation. Background technique [0002] In 1985, Galinski discovered the compensatory solute Ectoine, namely 1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid (1,4,5,6-tetrahydro-2 -methyl-4-pyrimidine carboxylic acid). Subsequent studies have found that Ectoine, as the main compensatory solute, is ubiquitous in moderately halophilic bacteria. Ectoine can protect enzymes, DNA, cell membranes and whole cells under adverse environments such as high temperature, freezing and drying. Therefore, Ectoine has great application value and broad application prospects in the fields of cosmetics, biological preparations, enzyme preparations and pharmaceuticals. [0003] In the past ten years, the process research of efficient production of Ectoine, first of all, is to solve the high-density culture of cells, the first techn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/12C12R1/01
Inventor 张苓花郎亚军
Owner DALIAN MARITIME UNIVERSITY
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