Method of screening strains for producing microorganism plastics

A technology for microorganisms and plastics, applied in the field of screening microbial plastic strains based on single-cell Raman spectroscopy, can solve the problems of poor specificity, time-consuming, cumbersome operation methods, etc., and achieve good signal-to-noise ratio and easy operation

Inactive Publication Date: 2015-08-12
GUANGXI ACAD OF SCI
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Problems solved by technology

These methods all need to confirm that the polyester substance contained in the cells is PHA by staining, and the operation process is complicated and time-consuming.
More specifically, the specificity of Sudan black slide staining for PHA is poor and the operation method is cumbersome, so it is not an ideal screening method; the specificity of Nile blue staining is strong, and the operation is relatively simple and fast compared with Sudan black slide staining, but The Nile blue slide staining method requires cell fixation and staining; the Nile blue colony staining method is the easiest method to operate in the staining method, but it is necessary to select an appropriate concentration of the staining solution for different microorganisms, which is difficult for a large number of unknown microorganisms. It is difficult to determine for screening, and Nile blue dye and other reagents added to the medium bring unknown damage to the strains to be selected, and cannot be used for direct screening

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Embodiment Construction

[0028] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments. This embodiment is only to illustrate the present invention more clearly, rather than to limit the present invention.

[0029] 1. Correlation Analysis of Colony Cell Raman Spectrum and Shake Flask Fermentation PHB Yield

[0030] 1. Experimental materials

[0031] Strains: Cupriavidus necator H16 strain.

[0032] Incline medium: yeast powder 10g / L, peptone 10g / L, beef extract 5g / L, NaCl 5g / L, agar 20g / L, pH 7.0.

[0033] Seed medium: yeast powder 10 g / L, peptone 10 g / L, beef extract 5 g / L, NaCl 5 g / L, pH 7.0.

[0034] Shake flask fermentation medium: Na 2 HPO 4 12H 2 O 9.0 g / L, KH 2 PO 4 1.5 g / L, MgSO 4 ·7H 2 O 0.2 g / L, CaCl 2 2H 2 O 0.02 g / L, NaHCO 30.5 g / L, ferric ammonium citrate 0.05 g / L, (NH4) 2 SO 4 2.0 g / L, trace element solution 1 mL. The pH value is 7.0, and it is sterilized at 115 ℃ for 20 minutes.

[0035] Trace ele...

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Abstract

The invention discloses a method of screening strains for producing microorganism plastics and belongs to the technical field of screening microbial strains. The method mainly comprises the following steps: by adopting the combination of laser tweezers and a single-cell Raman technique, judging the ability of microorganism cells in synthesizing the microorganism plastics, and further separating and purifying the strains for producing the microorganism plastics with high yield. The method is simple and convenient to operate and free from dyeing, and detection can be directly performed in aseptic water; the method is also capable of rapidly and intuitively screening the strains for producing the microorganism plastics without damaging the detected cells serving as samples, thereby being a simple, convenient, efficient and reliable method which is worthy of popularization and capable of screening the strains for producing the microorganism plastics with high yield.

Description

technical field [0001] The invention belongs to the technical field of microbial strain screening, and in particular relates to a method for screening microbial plastic-producing strains based on single-cell Raman spectroscopy. Background technique [0002] Plastic products play an important role in human's daily life and industrial and agricultural production. Ordinary plastics mainly composed of synthetic resins have caused serious environmental problems because they are difficult to degrade. It is imminent to research and develop new biodegradable bioplastics. Among the many biodegradable materials, the microbial plastic produced by microbial fermentation - poly β-hydroxyalkanoate (PHA) not only has the mechanical properties and processing properties of ordinary polymer materials, but also has biodegradability, Biocompatibility, piezoelectricity, optical activity and other incomparable characteristics of ordinary polymer materials. PHA has broad application prospects in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04G01N21/65
Inventor 王桂文彭立新王忠文冯旻陶站华
Owner GUANGXI ACAD OF SCI
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