Pseudomonas putida WP07, preparation method and purpose

A technology of Pseudomonas putida, cgmccno.17760, applied in the field of microorganisms, can solve the problems of complex swill oil composition, complex process and high cost, achieve resource utilization and harmless treatment, simple screening process, low cost effect

Active Publication Date: 2019-11-12
INST OF URBAN ENVIRONMENT CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Invention patent 201510203237.4 discloses a process for synthesizing PHA from kitchen waste oil using high-efficiency bacteria. Cupriavidus necator (DSM428) is used to cultivate waste kitchen oil as a carbon source to synthesize PHA from kitchen waste oil. Obtain PHA, the hookworm copper greedy bacteria that this method adopts, its technological process is relatively complicated, and cost is higher
And because the composition of swill oil is complex, the method of converting and synthesizing PHA with it as a single carbon source is still insufficient

Method used

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  • Pseudomonas putida WP07, preparation method and purpose
  • Pseudomonas putida WP07, preparation method and purpose
  • Pseudomonas putida WP07, preparation method and purpose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: Domestication and screening of Pseudomonas putida WP07 bacterial strain

[0037] The soil from the smoke exhaust outlet of the canteen was selected, and the Pseudomonas strains that transformed and synthesized PHA using swill oil as a single carbon source were obtained through domestication with high concentration of swill oil and selective selection with Nile blue. Strain acclimatization and screening steps include:

[0038] (1) Acclimatization of strains: first, add swill oil to the inorganic salt medium at a ratio of 20g / L, then obtain a soil sample from the soil at the smoke exhaust outlet of the canteen and add it to the above-mentioned inorganic medium at a ratio of 5g / L. Cultivation and acclimatization; the acclimatization conditions of the strain are temperature 20°C-30°C, rotation speed 200-300rpm; every 5 days, continue to add swill oil to the inorganic salt medium at a ratio of 5g / L until the swill oil in the inorganic salt medium is added Afte...

Embodiment 2

[0044] Example 2 Effect of Fermentation Temperature on Pseudomonas putida WP07 Transformation and Synthesis of PHA Efficiency

[0045] Insert the Pseudomonas putida WP07 obtained in Example 1 into the seed culture medium, the inoculum size is 1% (V / V), at 30°C, 200rpm number of revolutions conditions are cultivated for 24h activation to obtain the activated Pseudomonas seed culture.

[0046] Add swill oil as a single carbon source to the inorganic salt medium at a ratio of 20g / L, and add 1:10 (w / w) tween 80 as an emulsifier at the same time; then the activated Pseudomonas seed culture solution Insert the inoculation amount of 3% (v / v) into the inorganic salt medium added with swill oil, control the culture temperature at 20°C, 25°C, 30°C, 35°C and 40°C, and cultivate under the condition of 300rpm 72h (result comparison see figure 1 ). Among them, the seed medium preparation method is to add 10g / L tryptone, 5g / L yeast extract and 10g / L sodium chloride to deionized water, adj...

Embodiment 3

[0051] Example 3 Influence of the pH value of the inorganic salt medium on the conversion and synthesis of PHA by Pseudomonas putida WP07

[0052] The Pseudomonas putida WP07 obtained in Example 1 was inoculated into the seed culture medium, and cultured for 24 hours at 30° C. and 200 rpm for activation to obtain an activated Pseudomonas seed culture solution.

[0053] Add swill oil as a single carbon source to the inorganic salt medium at a ratio of 20g / L, and add 1:10 (w / w) tween 80 as an emulsifier at the same time; then add the seed culture solution to 3% (v / v ) was inserted into the inorganic salt medium added with swill oil, and the culture temperature was controlled to be 30° C. and the rotation speed was 300 rpm to cultivate for 72 hours. Among them, the seed medium preparation method is to add 10g / L tryptone, 5g / L yeast extract and 10g / L sodium chloride to deionized water, adjust the pH value with 0.1N HCl and 0.1N NaOH solution after mixing is 7.0.

[0054] Inorgan...

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Abstract

The invention discloses pseudomonas putida WP07, a preparation method and purpose. The pseudomonas putida WP07 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.17760. By a microbiological method, pigwash oil is converted into biodegradable plastic PHA which has environment affinity, so that resource utilization and innocent treatment of the pigwash oil can be realized, the preparation cost of the PHA can also be reduced, and the pseudomonas putida WP07 has favorable application prospects.

Description

technical field [0001] The invention relates to the field of microbes, in particular to Pseudomonas putida WP07, a preparation method and use. Background technique [0002] Polyhydroxyalkanoate (PHA) is a high molecular polymer that is stored as an intracellular carbon source and energy in the unbalanced growth state of microorganisms with excess carbon source and insufficient nitrogen source. It exists widely in microorganisms in the form of hydrophobic particles. in cells. Because it has similar material properties to traditional petroleum-based plastics such as polyethylene and polypropylene, and can be synthesized from renewable energy sources such as carbohydrates and fatty acids, its structure is diverse and can be completely degraded into The ecological cycle of nature has been paid more and more attention. In addition, PHA is synthesized in bacteria, which ensures its stereospecificity (all chiral carbon atoms in the main chain are R-configuration), which is very i...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/36C12N1/04C12P7/62C12R1/40
CPCC12N1/04C12N1/20C12N1/36C12P7/625C12N1/205C12R2001/40
Inventor 唐晓达汪印余广炜
Owner INST OF URBAN ENVIRONMENT CHINESE ACAD OF SCI
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