Poly-3-hydroxy propionic acid copolymer and production method thereof

A polyhydroxy fatty acid and gene technology, applied in the field of genetic engineering, can solve problems such as low melting point and opacity

Active Publication Date: 2014-09-17
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Poly 3-hydroxypropionic acid also has certain defects in properties, such as low melting point, opacity caused by crystallization, etc., which limit its application ...

Method used

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  • Poly-3-hydroxy propionic acid copolymer and production method thereof
  • Poly-3-hydroxy propionic acid copolymer and production method thereof
  • Poly-3-hydroxy propionic acid copolymer and production method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Construction of Genetically Engineered Bacteria

[0043] By integrating the glycerol dehydratase gene (dhaB123) (dhaB1Gene ID: 7947197; dhaB2Gene ID: 7947198; dhaB3Gene ID: 7947200), glycerol dehydratase reactivating enzyme gene (gdrAB) (gdrA Gene ID: 6936977; gdrB Gene ID: 6938011 ) Escherichia coli overexpressed propionaldehyde dehydrogenase gene (pduP, Gene ID: 1253572), polyhydroxy fatty acid synthase gene (phaC, Gene ID: 4250156), β-ketoacyl-CoA thiolase gene (bktB , Gene ID: 4248815), acetoacetyl-CoA reductase gene (phaB 1 , Gene ID: 4249784), the propionyl-CoA synthetase gene (pcs') realizes the synthesis of P(3HP-co-3HV) with glycerol as the carbon source, and the synthesis of poly 3-hydroxypropionate-co-3-hydroxypentane by glycerol Acid metabolic pathways such as figure 2 shown.

[0044] 1. Gene integration

[0045] After the genes dhaB and gdrAB are cloned, the dhaB and gdrAB are integrated into the Escherichia coli genome through gene integrati...

Embodiment 2

[0065] Example 2 Production of P(3HP-co-3HV) by fermentation

[0066] The production of P(3HP-co-3HV) was investigated by adjusting the amount of inducer and the use of organic nitrogen source.

[0067] 2.1) 0.5mM IPTG and 0.02% arabinose induced

[0068] Change the nitrogen source of the M9 medium to 5g / L yeast powder, peptone or beef extract, inoculate activated Escherichia coli at a ratio of 1:100, when the OD 600 When the range of 0.6-0.8 was reached, the inducer IPTG and arabinose were added to the final concentrations of 0.5mM and 0.02%, respectively, and P(3HP-co-3HV) was extracted after continuing to culture for 48h.

[0069] When the nitrogen source was beef extract, P(3HP-co-3HV) accounted for 12.94% (w / w) of dry cell weight, and the yield was 0.45g / L, which was the highest. The yield was the lowest when the nitrogen source was peptone, P(3HP-co-3HV) accounted for 6.53% of the dry cell weight, and the yield was 0.24g / L. When the nitrogen source was yeast powder, P...

Embodiment 3

[0074] The P(3HP-co-3HV) and poly-3-hydroxypropionic acid samples obtained in the present invention are washed with ethanol and dried in vacuum, and the melting point and crystallinity are measured by differential scanning calorimetry, and the temperature setting range is -80°C to 200°C. °C, the nitrogen flow rate is 50ml / min. The melting point of P(3HP-co-3HV) increased from 76°C of poly-3-hydroxypropionic acid to 147°C, and the crystallinity decreased from 74% of poly-3-hydroxypropionic acid to 35%.

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Abstract

The invention discloses a poly-3-hydroxy propionic acid copolymer and a production method thereof and belongs to the technical field of genetic engineering. According to the poly-3-hydroxy propionic acid copolymer and the production method thereof, a glycerol dehydratase gene and a glycerol dehydratase re-activating enzyme gene are integrated with a host strain genome by a gene integration technology, a polyhydroxy fatty acid synthase gene, a propionaldehyde dehydrogenase gene, a beta-ketoacyl coenzyme A thiolase gene, an acetoacetyl coenzyme A reductase gene and a propionyl coenzyme A synthetase gene are introduced, and a recombinant gene engineering strain has the ability of biologically synthesizing poly-3-hydracrylic acid-co-3-hydroxyvaleric acid. According to the poly-3-hydroxy propionic acid copolymer and the production method thereof, the poly-3-hydracrylic acid-co-3-hydroxyvaleric acid is obtained in a biosynthesis manner for the first time; compared with poly-3-hydracrylic acid, the obtained poly 3-hydracrylic acid-co-3-hydroxyvaleric acid has a higher melting point and lower crystallinity, has good degradability and can serve as a packaging material, a medical implant material, a drug sustained-release material and an electrochemical material.

Description

technical field [0001] The invention relates to a poly-3-hydroxypropionic acid copolymer and a production method thereof, belonging to the technical field of genetic engineering. Background technique [0002] The fossil energy crisis is becoming more and more serious, and the manufacture of biofuels has become a research focus. Biodiesel is the most important type of bioliquid fuel. Accompanied by the mass production of biodiesel, its by-product glycerol accumulates in large quantities. According to preliminary estimates, 1 ton of crude glycerin will be produced for every 10 tons of biodiesel produced. The further growth of biodiesel has caused the price of glycerin to continue to decrease. Making full use of low-priced glycerin to produce high value-added products can bring huge economic and environmental benefits, and has received more and more attention. [0003] Polyhydroxyalkanoate is a bioplastic obtained by microbial fermentation, which can be synthesized from che...

Claims

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Application Information

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IPC IPC(8): C12P7/62C12N1/21C12N15/63C12R1/19
Inventor 咸漠赵广冯新军
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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