Method for effectively producing 1,3-propanediol by modifying permeability of cell membrane

A propylene glycol and permeability technology, applied in 1 field, can solve the problems of long production cycle, low bacterial volume, low production intensity, etc., and achieve the effects of improving production efficiency, maintaining cell growth, and increasing permeability

Inactive Publication Date: 2008-07-30
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0012] However, in the existing processes, there are products and by-products that inhibit cell growth and cell catalytic activity, resulting in low bacterial mass in the fermentation process, long production cycle, and low production intensity.

Method used

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  • Method for effectively producing 1,3-propanediol by modifying permeability of cell membrane

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[0036] Bacteria: Klebsiella pneumoniae

[0037] Solid slant medium: beef extract 3g / L, MgSO 4 ·7H 2 O 0.2g / L, peptone 5g / L, NaCl 10g / L, agar 20g / L, glycerin 20g / L.

[0038] Seed medium: yeast extract 2g / L, MgSO 4 ·7H 2 O 0.2g / L, citric acid 0.42g / L, K 2 HPO 4 1.6g / L, K 2 HPO 4 3.4g / L, NH 4 Cl 3.0g / L, glycerol 30g / L, CaCl 2 0.5g / L.

[0039] Fermentation medium: glycerol 20-65g / L, K 2 HPO 4 ·3H 2 O 1.36g / L, NH 4 Cl 3.0g / L, yeast extract 2.0g / L, MgSO 4 ·7H 2 O 0.2g / L, CaCl 2 2.0g / L iron solution 1mL / L, inorganic ion solution 2mL / L, V B12 Solution 1mL / L, cobalt solution 1mL / L. Inorganic ions contain: CuCl 2 , H 3 BO 3 , MnCl 2 , Na 2 MoO 4 , NiCl 2 , ZnSO 4 , the content of trace elements is about 0.01-50mg / L.

[0040] Secondary seed preparation: take the fresh solid slant strain of Klebsiella pneumoniae, inoculate it in the seed medium, cultivate it on a shaker at 35°C at 150rpm for 24h, then inoculate it into the seed medium, and cultivate it at 35°...

Embodiment 1

[0044] 400ml of fermentation medium containing 45g / L industrial glycerin was placed in a 1L shake flask, and a volume of 40ml of secondary seeds was inserted, and the temperature was 37°C, and the shaker was cultivated at 180rpm. Add the concentration of 0%, 0.05%, 0.1%, 0.5%, 0.6%, 0.8%, 1% OP-10 respectively in the fermentation 8h, take samples at intervals, measure the cell concentration by spectrophotometer turbidimetry, the wavelength is 650nm , the cell concentration, that is, the amount of bacteria, is expressed as OD; the concentration of 1,3-propanediol is analyzed by gas chromatography. 24 hours to end fermentation.

[0045] Embodiment 1 has studied in fermenting glycerol to produce 1,3-propanediol technology, adds non-ionic surfactant OP-10, concentration is from 0~1.0% scope, different OP concentration is to Klebsiella pneumoniae growth and 1,3 - Effect of Propylene Glycol Yield. Figure 1 shows that at low concentrations, OP-10 mainly exhibits a promoting effect ...

Embodiment 2

[0048] 400ml of fermentation medium containing 50g / L industrial glycerol was placed in a 1L shake flask, and a volume of 40ml of secondary seeds was inserted, and the temperature was 37°C, and the shaker was cultivated at 180rpm. 0.1% OP-10 was added at 4h, 8h, 12h and 16h of fermentation, and the fermentation was finished in 24 hours. The concentration of 1,3-propanediol was analyzed by gas chromatography.

[0049]

[0050] Table 1 Fermentation 4h, 8h, 12h, 16h respectively added concentration of 0.1% OP-10 shake flask number

[0051] As can be seen from Figure 3, the addition of OP at different stages of fermentation has a greater impact on the output of 1,3-propanediol, and the effect of adding OP in 8-16 hours is the best, and the maximum output of 1,3-propanediol has increased by 12%. .

[0052] Through the above experiments, it can be proved that in the process of producing 1,3-propanediol by fermentation of glycerol, the method of increasing the permeability of the...

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Abstract

The invention relates to a method of effectively producing 1, 3-propylene glycol by changing the membrane permeability, which adds nonionics to change the membrane permeability, effectively promotes the cell growth, and increases the concentration of 1, 3-propylene glycol which is the finished product of fermentation. The utility model has the technical process that as follows: adding the second-class germ culture into the fermentation culture; adding nonionics which can improve the membrane permeability to the fermentation culture; the nonionics reduce the resistance of the oxygen and nutrient substance from entering the cell, promote the cell growth, enhance the secreting of metabolite to the extra-cellular area, reduce the accumulation of metabolite in cell, and facilitate the elimination of inhibition of product and sub product, in particular the inhibition of cell growth and cell catalytic activity by 1, 3-propylene glycol. The concentration of the fermentation product--1, 3-propylene glycol is increased by 10 to 70 percent. With simple technical process and low production cost, the invention is easy to be industrially applied.

Description

technical field [0001] The invention is a method for preparing 1,3-propanediol through biotechnology, in particular to a method for adding nonionic surfactants to change the permeability of cell membranes and effectively increasing the concentration of 1,3-propanediol, which belongs to the biocatalysis of 1,3-propanediol The field of synthetic technology. Background technique [0002] In organic synthesis reactions, 1,3-propanediol, as a bifunctional organic molecule, has a wide range of applications. It can replace ethylene glycol and butanediol, and can be used as a monomer for synthesizing polyester and polyurethane, as well as a solvent, antifreeze or protective agent, etc. It can also be used for synthesizing medicine and as an intermediate in organic synthesis. [0003] Its main purpose is to manufacture the important monomer raw material of new polyester fiber polytrimethylene terephthalate (PTT) with excellent performance. PTT is a high-grade clothing fiber materia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/18C12R1/22C12R1/01
Inventor 吴敏马全红
Owner SOUTHEAST UNIV
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