Rhodococcus pyridinivorans and application thereof in production of PHBV

A technology of Rhodococcus pyridineophilus and bacterial strains, applied in the field of Rhodococcus pyridinephilus and its application in the production of PHBV, can solve the problems of inadequate recycling, environmental pollution, waste of resources, etc., and achieve good application prospects, physical properties and The effect of excellent thermodynamic performance

Active Publication Date: 2020-12-11
THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The application of plastics has brought great convenience to people's lives. Due to excessive use and inadequate recycling, it has also caused serious waste of energy, resources and environmental pollution.

Method used

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  • Rhodococcus pyridinivorans and application thereof in production of PHBV
  • Rhodococcus pyridinivorans and application thereof in production of PHBV
  • Rhodococcus pyridinivorans and application thereof in production of PHBV

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Experimental program
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Effect test

Embodiment 1

[0030] Isolation and identification of strains:

[0031] The obtained Pacific deep-sea sediments were diluted with sterile water and spread on 2216E solid agar plates, and a single colony was picked for streaking and purification to obtain a strain that could produce white particles in the bacteria under a transmission electron microscope and named it P23 . The transmission electron microscope photos of the bacteria strain P23 are as follows: figure 1 As shown, it can be seen that the white particles in the bacteria are PHBV. The genome of the strain was extracted, the whole genome was sequenced, and the 16S rDNA sequence was obtained as shown in SEQ ID NO:1. Comparing the 16S rDNA at https: / / www.ezbiocloud.net / website, it was found that its similarity with the standard strain Rhodococcuspyridinivorans DSM 44555 was 99.93%, indicating that the strain P23 was Rhodococcus pyridinivorans (Rhodococcuspyridinivorans). At the same time, a phylogenetic tree based on the 16S rDNA ...

Embodiment 2

[0034] PHBV was synthesized by fermenting terephthalic acid as a substrate by using Rhodococcus pyridinivorans P23:

[0035] Rhodococcus pyridinivorans P23 was inoculated into 2216E liquid medium for test tube fermentation culture, and the cultured seeds were inoculated into the fermentation medium at 1%. The composition of the fermentation medium is as follows: every liter contains 26.0 g of sodium chloride, 5.0 g of magnesium chloride hexahydrate, 1.06 g of calcium chloride, 4.0 g of sodium sulfate, 0.3 g of ammonium chloride, 0.1 g of potassium dihydrogen phosphate, and 0.1 g of potassium chloride 0.5g, 1ml of trace elements, 10g of terephthalic acid, dilute to 1 liter with water. The pH value was adjusted to 7.0, the culture temperature was 30° C., the rotation speed of the shaker was 180 rpm, and the fermentation time was 48 hours. The fermentation was tested with a 500mL Erlenmeyer flask, and the filling volume was 150mL. After the fermentation, the bacteria were colle...

Embodiment 3

[0037] Characterization of physical properties of PHBV synthesized by fermentation of Rhodococcus pyridinivorans P23:

[0038] Take 10mg of PHBV sample for differential scanning calorimetry analysis, first pre-cool the sample to -100°C, heat it up to 200°C at a speed of 10°C / min and record, quench to -100°C and again at a speed of 10°C / min A temperature rise to 200°C was performed and recorded. The melting temperature of PHBV was found to be 101.0°C and the glass transition temperature was -14.4°C. The PHBV was made into a dumbbell-shaped diaphragm with a thickness of 0.6mm and a length and width of 35×6mm for tensile mechanics experiments. It was found that the tensile strength of PHBV was 1.0Mpa and the elongation at break was 15%. 10 mg of PHBV sample was dissolved in chloroform, and the molecular weight was determined and analyzed by gel permeation chromatography. The weight average molecular weight was 820 kDa.

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Abstract

The invention discloses a strain of rhodococcus pyridinivorans and application thereof in production of PHBV. The strain rhodococcus pyridinivorans (Rhodococcus pyridinivorans)P23 is preserved in theChina centre for type culture collection (CCTCC) on August 7th, 2019; the address is Wuhan University, Wuhan, China; and the preservation number is CCTCC NO:M2019609. The strain rhodococcus pyridinivorans (Rhodococcus pyridinivorans)P23 disclosed by the invention can be used for efficiently synthesizing a copolymer PHBV of 3-hydroxybutyric acid and 3-hydroxyvaleric acid in a mineral salt culture medium by taking terephthalic acid or terephthalate as a substrate. The monomer content of 3-hydroxyvaleric acid in the PHBV synthesized by the strain rhodococcus pyridinivorans (Rhodococcus pyridinivorans)P23 in the invention is more than 60%; furthermore, the weight-average molecular weight of the PHBV is higher than 600 kDa; and the PHBV has excellent physical properties and thermodynamic properties, and has a good application prospect.

Description

technical field [0001] The invention relates to the technical field of degradable plastics, in particular to a strain of rhodococcus pyridineophilus and its application in the production of PHBV. Background technique [0002] Plastics are widely produced and used because of their light weight, strength, ease of processing, low price, and good insulators. According to "Plastic Europe" statistics, the world produced 348 million tons of plastic in 2017, and China is the world's largest plastic producer, accounting for 29.4% of the world's plastic production (https: / / www.plasticseurope.org / en) . The application of plastics has brought great convenience to people's lives, but due to reasons such as excessive use and inadequate recycling, it has also caused serious waste of energy, resources and environmental pollution. As early as December 2007, China issued the "Plastic Restriction Order", which clearly stipulates: "From June 1, 2008, the production, sale and use of plastic sh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/62C12R1/01
CPCC12P7/625C12N1/20C12R2001/01C12N1/205
Inventor 郭文斌邵宗泽
Owner THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES
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