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Pseudomonas and application thereof to degrading macromolecular synthetic plastics

A technology of Pseudomonas bacteria, colonies, used in the field of microbial biotechnology and environmental biology

Inactive Publication Date: 2011-09-14
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Isolation and screening of highly efficient degrading strains is a necessary resource guarantee for both the research on the mechanism of compounds and the construction of biological circulation systems, and it is even more important to screen strains that degrade a variety of polymers for research and production. Has important theoretical and practical significance, and there are few reports

Method used

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  • Pseudomonas and application thereof to degrading macromolecular synthetic plastics
  • Pseudomonas and application thereof to degrading macromolecular synthetic plastics
  • Pseudomonas and application thereof to degrading macromolecular synthetic plastics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Screening of degrading strains

[0027] According to literature reports, the number of microorganisms that degrade synthetic polyesters in nature has the following relationship: poly-β-hydroxybutyric acid-degrading bacteria > polycaprolactone-degrading bacteria > polylactic acid-degrading bacteria. Therefore, in order to obtain microorganisms that can degrade all three compounds, polylactic acid was first used as the sole carbon source for screening.

[0028] Place environmental samples in polylactic acid liquid medium for enrichment and spread them on emulsified plates, culture them upside down in a 30°C incubator, observe the growth of strains and the formation of transparent circles in 15-30 days, and screen for growth on plates And form the strains with obvious transparent hydrolysis circle. Screening media components and production methods are as follows:

[0029] Basic medium: 250mg yeast extract, 1g (NH 4 ) 2 SO 4 , 100mg NaCl, 200mg MgSO 4 ·7H 2...

Embodiment 2

[0034] Example 2 Degradation of polymer film by Pseudomonas sp.DS1001

[0035] Cut the film of polylactic acid, poly-β-hydroxybutyric acid or polycaprolactone to a size of 2cm*2cm, weigh it with an analytical balance, wipe the surface with 75% ethanol, sterilize it with ultraviolet light, and then put it into a sterile basic medium as the only carbon source for bacterial culture. Pseudomonas sp.DS1001 was inoculated into the culture medium with 10% inoculum amount, and cultured in shake flasks at 37°C and 150 rpm. At the same time, a bottle of culture medium without bacteria was placed under the same conditions as a control for membrane degradation. Samples were taken regularly, and the film was dried to a constant weight and then weighed to calculate the weight loss rate.

[0036] In the experiment of degrading the polylactic acid film, after 15 days of cultivation, the weight loss rate of the polylactic acid film reached 55%, while the control film did not lose weight, indica...

Embodiment 3

[0038] Example 3 Preparation of crude enzyme solution of Pseudomonas sp.DS1001 and analysis of its enzymatic hydrolysis product

[0039] Pseudomonas sp.DS1001 was cultured by shaking flask fermentation, and polylactic acid, poly-β-hydroxybutyric acid and polycaprolactone were used as inducers to induce their respective degrading enzymes. After 3 days, the supernatant was collected by centrifugation at 12000rpm, which was fermented crude enzyme liquid. Mix the induced fermented crude enzyme liquid with their respective substrates (polylactic acid, poly-β-hydroxybutyric acid, and polycaprolactone) emulsion, and incubate at 40°C for 15 minutes (polylactic acid reacts slowly, and the incubation time can be extended to 1-6 hours), centrifuge at 12000rpm to take the supernatant reaction solution, and use LDI-1700 laser desorption ionization time-of-flight mass spectrometer to measure the degradation products therein. The working parameters of the instrument are: mass range: 0-800m / ...

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Abstract

The invention belongs to the fields of microbial biotechnology and environmental biotechnology, particularly relates to pseudomonas and application thereof to degrading some macromolecular synthetic plastics. The pseudomonas is Pseudomonas sp. DS1001; a colony of the pseudomonas is circular and yellow white and has a smooth surface and a tidy edge; the thallus is in the shape of a short rod and is gram negative, has flagellum and does not have spore or capsule; a macromolecular compound such as polylactic acid (PLA), poly-beta-hydroxybutyric acid (PHB), polycaprolactone (PCL) and the like can be used as a unique carbon source for growth; and the pseudomonas has a high-efficiency degradation effect on the compounds. The pseudomonas can be applied to treatment and biological recycling of plastic wastes taking the macromolecular material as a main component.

Description

technical field [0001] The invention belongs to the field of microbial biotechnology and environmental biotechnology, and specifically relates to a strain of Pseudomonas sp. applications in plastics. Background technique [0002] Since the 19th century, chemically synthesized plastics based on petroleum have been widely used due to their light weight, low price and excellent performance, and have become indispensable polymer materials in modern society. Four pillars. However, with the rapid development of the plastics industry, more and more problems have emerged. The inherent non-degradability of plastics has caused serious "white pollution". The total amount of plastic waste dumped around the world is as high as tens of millions of tons every year. Although the traditional plastic waste treatment method based on terminal treatment can reduce the impact of plastic waste to a certain extent, it is expensive and easy to cause secondary pollution. In this case, the developm...

Claims

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Application Information

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IPC IPC(8): C12N1/20A62D3/02C12R1/38A62D101/28
Inventor 李凡陈珊王战勇王岩
Owner NORTHEAST NORMAL UNIVERSITY
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