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A method for cell surface modification based on oxidative self-polymerization of plant polyphenol tannic acid

A polyphenol tannic acid, cell surface technology, applied in the field of microorganisms, can solve the problems of irreversible experimental results, long experimental period of physical and chemical methods, low experimental safety factor, etc., and achieve the effects of mild experimental conditions, simple operation and avoiding pollution.

Active Publication Date: 2021-10-15
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the means of genetic engineering are limited by the metabolic pathways of the cells themselves, and the experimental cycle of physical and chemical methods is too long, the experimental results are irreversible, and the experimental safety factor is low.

Method used

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  • A method for cell surface modification based on oxidative self-polymerization of plant polyphenol tannic acid
  • A method for cell surface modification based on oxidative self-polymerization of plant polyphenol tannic acid

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Embodiment 1

[0027] In this embodiment, the method of modifying the cell surface based on the oxidative self-polymerization of plant polyphenol tannic acid prepared is as follows:

[0028] Select yeast as an illustration example, collect the Yarrowia lipolytica cells cultivated to the logarithmic phase with sterile centrifugal acid centrifugation, divide into different concentrations of tannic acid solutions prepared with different pH PBS solutions, shake on the shaker The reaction causes tannic acid to oxidize and self-polymerize on the cell surface to form a tight protective shell.

[0029] Specific steps include:

[0030] 1. Prepare PBS solution with pH=6.0~8.0. Liquid A: 0.05mol / LNa 2 HPO 4 12H 2 O solution, weigh 17.907 g of disodium hydrogen phosphate dodecahydrate, and fully dissolve it with 1000 ml of distilled water. Liquid B: 0.05mol / LKH 2 PO 4 Solution, weigh potassium dihydrogen phosphate, 6.8045g, fully dissolve with 1000ml distilled water. A and B liquids are mixed in...

Embodiment 2

[0037] In this embodiment, the method of modifying the cell surface based on the oxidative self-polymerization of plant polyphenol tannic acid prepared is as follows:

[0038] Centrifuge the Yarrowia lipolytica cultured to the logarithmic phase and collect it with a sterile centrifuge tube, divide it into the tannic acid solution prepared with PBS buffer, and shake it on the shaker to make the tannic acid on the cell surface Oxidative self-polymerization occurs to form a tight protective shell.

[0039] Specifically include the following steps:

[0040] 1. Transfer Yarrowia lipolytica ATCC201249 from the storage tube to the YPD solid medium, activate by streaking, pick a single colony and insert it into the YPD test tube for activation for 6-24 hours, then transfer to the YPD shake flask, at 30° C. Cultivate at 225rpm to OD=0.8~1.0, and collect bacteria by centrifugation at 3000rpm.

[0041] 2. Configure 95% 0.05mol / L Na in PBS solution with pH=8.04 2 HPO 4 12H 2 O and 5...

Embodiment 3

[0047] In this embodiment, the method of modifying the cell surface based on the oxidative self-polymerization of plant polyphenol tannic acid prepared is as follows:

[0048] The Escherichia coli cultured to the logarithmic phase was collected by centrifugation in a sterile centrifuge tube, divided into tannic acid solution prepared with PBS solution, and reacted in a shaking table to make tannic acid oxidize and self-polymerize on the cell surface to form A tight protective shell.

[0049] Specifically include the following steps:

[0050] 1. Transfer Escherichia coli ATCC 8739 from the storage tube to the LB solid medium, activate by streaking, pick a single colony and insert it into the LB test tube for activation for 6-12 hours, then transfer to the LB shaker flask, at 37°C, Cultivate at 225rpm to OD=0.8~1.0, and collect the bacteria by centrifugation at 2500rpm.

[0051] 2. Configure 95% 0.05mol / L Na in PBS solution with pH=8.04 2 HPO 4 12H 2 O and 5% of 0.05mol / L ...

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Abstract

This paper discloses a method for modifying the cell surface based on the oxidative self-polymerization of plant polyphenol tannic acid. The enriched cells are placed in a tannic acid solution prepared by phosphate buffered saline (PBS), and shaken in a shaking table for 30 minutes. ~6h makes tannic acid oxidize and self-polymerize on the surface of the cell membrane to form a layer of tightly bound polymers, so that the whole cell can maintain activity for a long time in a complex fermentation environment and improve the tolerance to the product. The reaction conditions required by the invention are mild and the experimental operation process is simple. The whole reaction is completed in one step in the alkaline fermentation solution, which avoids pollution to the environment, and the tannic acid has a wide range of sources and exists in plant roots, stems, bark and various Chinese herbal medicines, and is cheap and suitable for large-scale industrialization.

Description

technical field [0001] The invention relates to a method for modifying cell surfaces based on oxidative self-polymerization of plant polyphenol tannic acid, which belongs to the technical field of microorganisms. Background technique [0002] With the rapid development of cell-based applications such as cell sensors, bioelectronic devices, and biocatalysis in recent years, this has put forward higher requirements for cells to maintain long-term viability and functionality in complex environments. Advanced molecular biology techniques are applied to modify cell metabolic pathways, while some physical and chemical methods are also used to improve cell stability and product separation, such as cell immobilization, medium optimization, ARTP mutagenesis, original For separation techniques, etc. However, the means of genetic engineering are limited by the metabolic pathways of the cells themselves, and the experimental period of physical and chemical methods is too long, the expe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12R1/645
CPCC12N1/16
Inventor 方艳王玉珍姜岷张展略马江锋信丰学章文明
Owner NANJING TECH UNIV
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