Method for transforming 1,3-propylene glycol into 3-hydroxypropionic acid by applying resting cells

A technology of hydroxypropionic acid and propylene glycol, applied in the field of bioengineering, can solve the problems of long production cycle, unstable bacterial species in fermentation method, and many side reactions, save energy and production costs, facilitate product separation and purification, and transform liquid components. simple effect

Active Publication Date: 2014-01-22
JIANGNAN UNIV
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  • Abstract
  • Description
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Problems solved by technology

The chemical synthesis method is very difficult to introduce functional groups at the carbon end, there are many side reactions, high process requirements, many by-products are difficult to separate and purify, the production cost is high, the operation is complicated, there are unsafe factors in the production process, and the output is low
[0004] The microbial fermentation method is mainly to construct engineering bacteria through genetic engineering. There are two main strategies: the production route using glucose as the substrate and the production route using glycerol as the substrate. Long cycle, cumbersome product purification process, low conversion rate, low yield, low product purity, and a considerable amount of 3-hydroxypropionate ether bond dimer, etc.

Method used

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  • Method for transforming 1,3-propylene glycol into 3-hydroxypropionic acid by applying resting cells
  • Method for transforming 1,3-propylene glycol into 3-hydroxypropionic acid by applying resting cells
  • Method for transforming 1,3-propylene glycol into 3-hydroxypropionic acid by applying resting cells

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Effect test

Embodiment 1

[0022] Example 1 High conversion of 1,3-propanediol performance, screening of microbial strains with high ability to produce 3-hydroxypropionic acid

[0023] Enrichment medium (g / L): glucose 20, yeast extract 5, peptone 2.

[0024] Plate screening medium (g / L): Glucose 20, yeast extract 5, agar 20, CaCO310 (added after sterilization alone), absolute ethanol 30 (added after the medium is sterilized).

[0025] Seed medium, fermentation medium (g / L): glucose 20, yeast extract 5, peptone 2, pH 6.0.

[0026] Analysis method: detection of 3-hydroxypropionic acid by HPLC, chromatographic column: BIORAD HPX-87H chromatographic column; mobile phase 0.005mol / L concentrated sulfuric acid; flow rate 0.6mL / min; detection temperature 60°C; detection wavelength 210nm.

[0027] Add the collected soil from different regions into sterile saline solution containing glass beads and shake for 1 hour, then let it stand still, take the supernatant and add it to the enrichment medium for static cult...

Embodiment 2

[0031] The impact of embodiment 2 different substrate concentrations on conversion

[0032]Centrifuge the cells cultivated in the fermentation medium for 32 hours, wash the cells twice with a phosphate buffer solution of pH 6.0, suspend the cells with a phosphate buffer solution of pH 6.0, and make the cell concentration 6g / L Add different amounts of substrate 1,3-propanediol to the bacterial suspension, so that the final concentrations are 10g / L, 20g / L, 30g / L, 40g / L, 50g / L, and catalyze at 30°C and 220rpm Sampling within 72 hours, the transformation solution was centrifuged to remove bacteria, filtered through a 0.22um aqueous phase membrane, and tested by HPLC to detect its yield, the yield can reach 9.6g / L, 18.9g / L, 27.7g / L, 35.9g / L L, 45.5g / L, the conversion rate is above 90% (attached Figure 5 ).

Embodiment 3

[0033] The influence of embodiment 3 different initial pHs on conversion

[0034] After centrifuging the bacteria cultured in the fermentation medium for 32 hours, use pH4.0, pH4.5, pH5.0, pH5.5, pH6.0, pH6.5, pH7.0, pH7.5, pH8.0 respectively After washing the cells twice with the buffer solution, suspend the cells with the buffer solution of the corresponding pH to make a cell suspension with a cell concentration of 6 g / L, add 20 g / L of 1,3-propanediol, and measure at 220 rpm at 30 °C. The amount of 3-hydroxypropionic acid produced after 6h of catalysis was taken as its initial catalytic rate. The catalytic rate is the highest at around pH 6.0, which can reach 1.1g / L*h. The acidic or alkaline pH will cause the catalytic rate to decrease, and the catalytic rate will decrease by about 40% at around pH 4.0 (attached Image 6 ).

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Abstract

The invention discloses a method for transforming 1,3-propylene glycol into 3-hydroxypropionic acid by applying resting cells, and belongs to the field of biological engineering. Through the method, firstly a rapid screening method is utilized to screen a microbial strain which has high transformation performance and high production capacity and is identified as acetobacter sp. JDB-71, and the resting cells of the strain are utilized for high-efficiency catalysis of 1,3-propylene glycol into 3-hydroxypropionic acid in an aqueous phase system. The obtained acetobacter sp. culture process is simple, the transformation efficiency is high, and the product yield is large; transformation solution components are simple, so as to be benefitial for separation and purification of the product; and the thalli concentration can be artificially adjusted to shorten the production time during the transformation process, and thus a lot of energy and production costs are saved.

Description

technical field [0001] The present invention relates to a method for converting 1,3-propanediol into 3-hydroxypropionic acid using resting cells, especially using a strain of Acetobacter and its resting cells to convert 1,3-propanediol into 3-hydroxypropionic acid The method belongs to the field of bioengineering. Background technique [0002] 3-hydroxypropionic acid (3-HP for short), also known as β-hydroxypropionic acid, is isomers with lactic acid, because it has a hydroxyl group and a carboxyl group at both ends of its carbon chain. A chemical intermediate with high value, such as alcohol or aldehyde can be obtained by reduction of carboxyl group, reacted with alcohol to obtain ester or converted into amide and its derivatives, aldehyde or diacid can be obtained by hydroxyl oxidation, dehydration can be converted into unsaturated compound, Polymerization to obtain biodegradable polymer materials, etc. In addition, the research data of Schwarz et al. showed that 3-hydrox...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/42C12R1/02
Inventor 诸葛斌吴金鑫宗红陆信曜宋健方慧英诸葛健
Owner JIANGNAN UNIV
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