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Bacillus safensis PLA1006 strain, screening method and application thereof

A technology of Bacillus suffolus and screening methods, applied in the field of microbial technology and environmental biology, can solve the problems of weak PLA substrate binding, low PLA degradation efficiency, lack of evolutionary process of microorganisms, etc.

Active Publication Date: 2022-06-24
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reason is that PLA is a non-natural synthetic compound that has appeared in recent decades, and microorganisms lack hundreds of millions of years of evolution, so the degradation efficiency of microorganisms in nature is relatively low for PLA
At present, most of the PLA degrading enzymes that have been isolated and identified originally existed in the natural environment without any evolution process, so the binding force to the PLA substrate is generally not strong.

Method used

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  • Bacillus safensis PLA1006 strain, screening method and application thereof
  • Bacillus safensis PLA1006 strain, screening method and application thereof
  • Bacillus safensis PLA1006 strain, screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Screening of degrading strains

[0035]The soil samples collected from the Fushun domestic waste landfill were placed in a liquid medium containing polylactic acid powder for enrichment, and then coated on an emulsified plate medium. In the case of formation, the strains that can grow on the plate and form a clear transparent hydrolysis circle were screened. The components of the screening medium and the preparation method are as follows:

[0036] The basal medium composition is: 4.54g KH 2 PO 4 , 11.94g Na 2 HPO 4 ·12H 2 O, 0.5 g MgSO 4 ·7H 2 O, 1 g NH 4 Cl, 0.005g CaCl 2 ·2H 2 O, 0.002g FeSO4 7H 2 O, 0.001gMnSO 4 ·H 2 O. 0.002g ZnSO 4 ·7H 2 O and 0.2g yeast powder. Dilute to 1L with deionized water, pH 6.8-7.0.

[0037] Polylactic acid liquid medium: Add 2 g of polylactic acid powder to the above basal medium.

[0038] Preparation of emulsification plate: 0.2g of polylactic acid was dissolved in 10ml of dichloromethane, the resulting soluti...

Embodiment 2

[0040] Example 2 Degradation of polylactic acid film by Bacillus safensis PLA1006 strain

[0041] The polylactic acid film was cut into a size of 3m×3cm, accurately weighed, wiped with 70% ethanol, irradiated with an ultraviolet lamp for 30 minutes, and placed in a sterile basic medium. The Bacillus safensis PLA1006 strain was inserted into the medium, 30° C., 150 rpm shaking culture, regular sampling, drying the film to a constant weight and then weighing, and calculating the weight loss rate.

[0042] In the experiment of degrading polylactic acid film, after 10 days of culture, the weight loss rate of polylactic acid film reached 15% ( Figure 4 ), and although the control film lost weight, the weight loss rate was lower than that of the experimental group, indicating that the degradation of the PLA film was mainly caused by the action of microorganisms or their products, rather than simple hydrolysis.

Embodiment 3

[0043] Example 3 Preparation of crude enzyme solution of Bacillus safensis PLA1006 strain and analysis of polylactic acid degradation products

[0044] Bacillus safensis PLA1006 was cultured in a polylactic acid liquid medium, the medium was the same as that of Example 1, and after 7 days of culture, the supernatant was collected by centrifugation at 12,000 rpm and concentrated by ultrafiltration, which was the crude enzyme liquid. The crude enzyme solution was mixed with the PLA emulsion, incubated at 40°C for 12 hours, centrifuged at 12,000 rpm to take the supernatant reaction solution, and the sub-membrane solution after ultrafiltration (molecular weight cut-off of 3,000 Daltons) was used to measure the degradation products using a laser desorption ionization time-of-flight mass spectrometer. . The measurement results( Figure 4 ) showed that polylactic acid enzymatic hydrolysis products were found to include lactic acid monomer (mass spectrometry peak 89) and lactic acid ...

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Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a bacillus safensis PLA1006 strain, a screening method and application of the bacillus safensis PLA1006 strain. The preservation number of the strain is CGMCC No.24188, the preservation date is December 24, 2021, the strain can grow by taking polylactic acid as a carbon source, and the specific degradation process comprises the following steps: (1) taking a dried polylactic acid film, wiping the surface with 70% ethanol, carrying out irradiation sterilization with an ultraviolet lamp, and putting into a sterile basal culture medium; and (2) inoculating the bacillus safensis PLA1006 strain into the basic culture medium, sampling regularly, and determining the weight loss condition of the film. The strain has protease production capacity and polylactic acid utilization capacity, polylactic acid powder, films and emulsion can be degraded by the bacillus safensis provided by the invention, and the strain can be applied to treatment of polylactic acid product wastes.

Description

technical field [0001] The invention belongs to the field of microbial technology and environmental biotechnology, in particular to a protease-producing Bacillus safensis PLA1006 strain and its application in the degradation of polylactic acid. Background technique [0002] Plastics are widely used for their low cost, light weight, corrosion resistance, stable performance and easy processing and molding. However, with the development of the plastics industry and the widespread use of disposable plastic products, the problem of environmental pollution caused by plastic waste is becoming more and more serious. Therefore, the replacement of traditional plastics by biodegradable plastics has become the development trend of the plastics industry. [0003] The ideal biodegradable plastic is a polymer material with excellent performance, which can be completely decomposed by environmental microorganisms after being discarded, and finally be inorganicized to become an integral part...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02B09B3/70C12R1/07B09B101/75
CPCC12N1/20C12N1/02B09B3/00Y02W30/62
Inventor 王战勇王玉俊吕淑霞林景卫
Owner SHENYANG AGRI UNIV
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