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Bacterium and enzyme mixed preparation containing Stenotrophomonas pavanii and application of bacterium and enzyme mixed preparation

A technology of Stenotrophomonas and maltophilia, applied in the field of microbiology

Active Publication Date: 2020-10-27
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many studies on the degradation of polyethylene terephthalate (PET) by multiple enzymes, but there are few studies on the synergistic degradation of polyethylene terephthalate (PET) by bacterial enzymes (mixed preparations)

Method used

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  • Bacterium and enzyme mixed preparation containing Stenotrophomonas pavanii and application of bacterium and enzyme mixed preparation
  • Bacterium and enzyme mixed preparation containing Stenotrophomonas pavanii and application of bacterium and enzyme mixed preparation
  • Bacterium and enzyme mixed preparation containing Stenotrophomonas pavanii and application of bacterium and enzyme mixed preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1: Screening and identification of Stenotrophomonas pavanii

[0061] 1. Screening of Stenotrophomonas pavanii

[0062] Specific steps are as follows:

[0063] 1. Ordinary screening

[0064] Taking the soil from the Taohuashan landfill in Wuxi City as a sample, take 1g of soil and add 9mL of normal saline, shake and enrich for 30min at 35°C and 180rpm; then take 1mL of the supernatant and dilute to 10 -4 、10 -5 、10 -6 , respectively take 200μL and dilute to 10 -4 、10 -5 、10 -6 The diluted solution was evenly spread on the inorganic salt solid medium containing 2g / L polyethylene terephthalate (PET), and placed in a 35°C incubator for constant temperature cultivation until colonies grew; the experimental results showed that after many Daily culture, no bacterial colonies appeared on the separation plate, this method can not effectively isolate the bacterial strain with the ability to degrade polyethylene terephthalate (PET).

[0065] 2. "PET induction cultu...

Embodiment 2

[0079] Embodiment 2: the preparation of bacterial enzyme mixed preparation

[0080] Specific steps are as follows:

[0081] (1) Activated strains: Spread Stenotrophomonas maltophilia JWG-G1 on an LB solid medium plate and culture at 37°C and 150rpm for 1 day to obtain a single colony;

[0082] (2) Preparation of seed solution: scrape the single colony obtained in step (1) and inoculate it into a glass test tube of 5 mL liquid LB medium, and cultivate it at 30° C. and 150 rpm for 12 hours to obtain the seed solution;

[0083] (3) Expansion cultivation: the seed liquid obtained in step (2) (viable bacteria number 1 × 10 8 CFU / mL) was transferred to LB liquid medium at a volume ratio of 1%, and cultivated to OD at 30°C and 150rpm 600 =1.0, obtain fermented liquid;

[0084] (4) Preparation of bacterial agent: the fermented liquid obtained in step (3) is centrifuged at 3600rpm for 10min, the supernatant is discarded, and then the inorganic salt liquid medium is added until the n...

Embodiment 3

[0086] Embodiment 3: the degradation ability of Stenotrophomonas maltophilia JWG-G1 to the intermediate phthalate (BHET) of polyethylene terephthalate in different addition amounts

[0087] With the bacterial agent Stenotrophomonas maltophilia JWG-G1 (viable bacteria number 2 * 10) that step (4) in the embodiment 2 obtains 9 CFU / mL) were added to the 2mg The reaction system was obtained in the inorganic salt liquid of BHET; the reaction system was placed at 37° C. and 150 rpm to react for 15 hours, and the amount of MHET and TPA generated and the remaining amount of BHET were measured by HPLC.

[0088] The analysis results are shown in Table 1. With the increase of the addition amount of Stenotrophomonas maltophilia JWG-G1, the degradation rate of BHET gradually increased; when the addition amount of Stenotrophomonas maltophilia JWG-G1 was 20%, The degradation rate of BHET reached the maximum value (37%); after that, the degradation rate of BHET remained unchanged with the in...

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Abstract

The invention relates to a bacterium and enzyme mixed preparation containing Stenotrophomonas pavanii and application of the bacterium and enzyme mixed preparation, and belongs to the technical fieldof microorganisms. The invention provides a novel method for degrading polyethylene terephthalate (PET) and a PET intermediate. By combining a specific strain and a specific biological enzyme, the bacterium and enzyme mixed preparation with specific degradation application is prepared, and the mixed preparation is a compound of Stenotrophomonas pavanii and cutinase; the bacterium and enzyme mixedpreparation provided by the invention is used to degrade PET and BHET intermediate, the degradation ability of the bacterium and enzyme mixed preparation is far better than that of using bacteria or enzymes alone. The bacterium and enzyme mixed preparation has a significant synergistic effect on high-crystallinity PET and the intermediate BHET in the degradation process, and has good promotion andapplication value in the field of the plastic degradation technology.

Description

technical field [0001] The invention relates to a bacterial enzyme mixed preparation containing Stenotrophomonas maltophilia and its application, belonging to the technical field of microorganisms. Background technique [0002] With the rapid development of the economy, the accumulation of plastic waste exceeds 320 million tons per year, and 60% of the plastic waste is polyethylene terephthalate (PET) plastic products. Since plastics are difficult to degrade, the global annual recycling rate of plastic products is only 14%, which makes plastic waste continue to accumulate in the environment and poses a serious threat to the ecology. Therefore, the degradation of polyethylene terephthalate (PET) is very critical to the control of plastic waste. [0003] Compared with traditional physical and chemical degradation technology, biodegradation technology is a technology that directly degrades plastics through degradable plastic strains. Due to its advantages of green, pollution-f...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/18A62D3/02A62D101/28
CPCC12N1/20C12N9/18A62D3/02C12Y301/01074A62D2101/28
Inventor 吴敬颜正飞陈晓倩黄青松顾冷涛
Owner JIANGNAN UNIV
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