Production technology of 1,3-propylene glycol by using glycerol fermentation method

A technology of propylene glycol and glycerin, which is applied in the direction of fermentation, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of low product concentration, low conversion rate of glycerin, and long production cycle, so as to reduce production costs and shorten fermentation time , Improve the effect of substrate conversion rate

Inactive Publication Date: 2013-06-12
XUZHOU KAIMIKE NEW MATERIALS
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AI Technical Summary

Problems solved by technology

[0004] At present, there are problems such as low product concentration, long production cycle and low conversion rate of glycerol in the production of 1,3-propanediol by biosynthesis. Therefore, using cheap and easy-to-obtain raw materials to prepare 1,3-propanediol under mild conditions is beneficial for improving 1, The preparation and expanded application of 3-propanediol and downstream products are of special significance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 1) Construction of bio-brick component Pet-sis

[0018] The genome of Micromonas eineua was extracted, and the primers required for synthetic PCR were designed according to the gene sequence of Micromonas eineua biosynthesis gene cluster (FJ160413) published by NCBI.

[0019] The upstream primer sisI1 is: CCC AAGCTT TCACACCGAGATCTGC (HindIII);

[0020] The downstream primer sisI2 is: CCG GAATTC ATGGAACCAGGACTTG (EcoR I).

[0021] To a 200 μL eppedoff tube, add 200 pmol of dNTPs, 20 μmol of upstream and downstream primers, about 1 ng of template DNA, 5 U Fast Pfu DNA polymerase, 10 μL of 5×Fast Pfu enzyme buffer, and then add sterile distilled water to a total volume of 50 μL. The reaction parameters of the Biometra TProfessional PCR instrument were pre-denaturation at 94°C for 5 min, denaturation at 94°C for 45 s, annealing at 66.8°C for 45 s, extension at 72°C for 90 s, and 30 cycles, and then extension at 72°C for 5 min to obtain PCR products.

[0022] Extract the p...

Embodiment 2

[0041] The composition and culture conditions of each medium are the same as above.

[0042] 1) Fermentation process:

[0043] Fermentation culture: Inoculate the expanded seed liquid with 9% inoculum in a 5L fermenter equipped with 2L fermentation medium, the stirring speed is 140rpm, and pass nitrogen into the fermenter during the fermentation process to maintain the anaerobic state of the fermentation system condition.

[0044] pH control: automatic adjustment to 6.4 with a concentration of 2mol / L NaOH solution.

[0045] Feeding control: according to the proportional relationship between glycerol consumption and lye consumption in batch fermentation, a mixed solution is prepared with the ratio (mass ratio) of glycerol (after conversion): NaOH=8:1, and the mixed solution is used to replace the simple solution when feeding. When adjusting the pH, add glycerol in a flow-in manner, and switch back to simple NaOH solution after feeding, so as to continue to consume the residua...

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Abstract

The invention discloses a production technology of 1,3-propylene glycol (PDO) by using a biological fermentation method, belonging to the technical field of biochemical engineering. The production technology comprises the following steps of: preparing 2'-dehydroxylase gene (sisII) from 3',4'-double dehydroxylase gene of sisI in micromonospora inyoensis through genetic recombination, DNA (deoxyribonucleic acid) rearrangement and the like; cloning to obtain sisII; constructing the sisII to a pET30a expression vector, and performing heterologous expression in E.coli BL21 to construct engineering bacteria BL21-sis for synthesizing 1,3-propylene glycol through glycerol dehydration; performing immobilization by a supramolecular self-assembly template method; fermenting to synthesize 1,3-propylene glycol in a glycerol-containing culture medium; and obtaining the product 1,3-propylene glycol through fermentation liquid filtration, organic solvent extraction and the like. The method for preparing 1,3-propylene glycol has the advantages of short production period, simple equipment, environmental friendliness, easiness in operation and control, cheap raw materials and the like, facilitates industrial production and has great practical value.

Description

technical field [0001] The invention belongs to the technical field of biochemical industry, and relates to a method for preparing 1,3-propanediol by a glycerol fermentation method. Background technique [0002] 1,3-propanediol is a chemical raw material with broad application prospects. It is mainly used as a monomer of polyester and polyurethane, as well as a solvent, antifreeze or protective agent. In addition, 1,3-propanediol can also be used in the synthesis of plasticizers, detergents, preservatives, and emulsifiers, and can also be used as components in products such as cosmetics, printer inks, cleaning agents, stabilizers, and fuel cell fuels, etc. additives to improve product performance. As an intermediate in medicine and organic synthesis, 1,3-propanediol can be used in industries such as food, cosmetics and pharmaceuticals. 1,3-propanediol is also used as an intermediate to produce polyol polyester and as a carbon chain extender, and can also be used to prepare...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/18C12N15/70C12N1/21C12R1/19C12R1/29
Inventor 蔡佩君赵云拾冲谭静萍李荣李成军赵明光
Owner XUZHOU KAIMIKE NEW MATERIALS
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