Method for screening PHA producing bacteria

A technology for producing bacteria and bromothymol blue, which is applied in biochemical equipment and methods, microbial determination/inspection, measuring devices, etc., and can solve the problems of time-consuming, labor-intensive, material waste, and high false positive rate

Pending Publication Date: 2021-07-06
陈宁远
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  • Claims
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Problems solved by technology

Most of them screen PHA-producing bacteria through single indicator screening or step-by-step screening of mu

Method used

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  • Method for screening PHA producing bacteria
  • Method for screening PHA producing bacteria
  • Method for screening PHA producing bacteria

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Embodiment Construction

[0014] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments, but it is not intended to limit the protection scope of the present invention.

[0015] A method for screening PHA producing bacteria, the steps are as follows:

[0016] 1) Preparation of medium: tryptone 1.5 g, yeast extract 1.5 g, glucose 10.0 g, sucrose 2.0 g, maltose 2.0 g, mannitol 1.0 g, sodium chloride 5.0 g, agar powder 20.0 g, bromothymol Blue 1.0 g, all reagents were dissolved in 800ml of pure water, stirred to dissolve, the volume was adjusted to 1000ml, the pH value was adjusted to 6.5-7.0, 121 ℃ for 20 minutes under high pressure, after cooling to about 60 ℃, add 1.0 g / L nilo Add the red stock solution to the sterile mixture above at a final concentration of 0.5 ug / ml, mix well and then pour it into a plate;

[0017] 2) Enrichment of PHA-producing bacteria: Take 5.0 g of river mud sample in a 250 mL Erlenmeyer flask, add 45 mL of s...

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Abstract

The invention discloses a method for screening PHA producing bacteria. The method comprises the following steps: 1) manufacturing a culture medium; 2) enriching PHA producing bacteria; and 3) screening the PHA producing bacteria: spraying a bromothymol blue indicator into a culture medium plate, marking bluing bacterial colonies around the bacterial colonies, placing the bacterial colonies under an ultraviolet lamp for observation, and determining that the bacterial colonies are PHA producing bacteria if orange fluorescence appears; and picking the single colonies by using a sterilization gun head, performing smearing, placing the single colonies under blue light of a fluorescence microscope for observation and verification, then, picking the single colonies, performing oscillation and enrichment in a shaking table for 2 hours, dipping a small amount of bacterial liquid by using an inoculating loop for line drawing separation on a new plate culture medium, after inverse culture at 37 DEG C for 24 hours, picking the single colonies and performing numbering, and performing repeated purification and separation in such a way. According to the method, two indicators are selected from numerous indicators for screening PHA producing bacteria, and a one-step screening method is adopted, so that time and materials are saved; meanwhile, the false positive rate can be reduced; and the period required by purification and separation can be shortened.

Description

technical field [0001] The invention belongs to the field of biology, in particular to a method for screening PHA-producing bacteria. Background technique [0002] PHA (Polyhydroxyalkanoate) is an intracellular polyester widely present in various microorganisms and other organisms. PHA has similar physical and mechanical properties to chemically synthesized plastics, and also has many excellent properties such as biodegradability, biocompatibility, optical activity, piezoelectricity, and gas barrier properties that synthetic plastics do not have. PHA is degradable in nature, causes little environmental pollution, is a renewable biological resource, is conducive to sustainable development, and has great application potential in many fields. For example, the physical and chemical properties similar to plastics can be used to make packaging materials or plastic products for daily consumption, and using them to replace the existing large-scale use of non-degradable plastics fro...

Claims

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Application Information

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IPC IPC(8): C12Q1/04G01N21/64
CPCC12Q1/04G01N21/6486
Inventor 陈宁远
Owner 陈宁远
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