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Polysaccharose film of brevibacterium and its use in bacteria preservisation

A pullulan polysaccharide and film technology, which is applied in the preservation of microorganisms and other directions, can solve the problems of long preservation period and high cost, and achieve the effects of low preservation cost, convenient use and small space occupation.

Inactive Publication Date: 2005-07-27
李世杰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bag Including actinomycetes and fungi, the metabolism is at a standstill Stunning state, easy to operate, long storage period The cost of equipment is relatively high, and the general grass-roots hard to reach Ultra-low temperature (in liquid nitrogen -196 °C) Metabolism is at a standstill, storage time Up to 20+ years The initial cost is high, and a stable economic conditions to ensure the supply of liquid nitrogen, If stored in liquid phase, exposure to It will explode when exposed to air. Needs good practice to use

Method used

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  • Polysaccharose film of brevibacterium and its use in bacteria preservisation
  • Polysaccharose film of brevibacterium and its use in bacteria preservisation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Rice starch is hydrolyzed to obtain a mixture of glucose and oligosaccharides, and the mixture is prepared into a culture medium, which is fermented by Aureobasidium into pullulan. The polysaccharide is separated by bacteria, decolorized, filtered, refined, concentrated to a concentration of 20g / 100ml, injected into the spray film head, the nozzle opening is 0.1mm, sprayed with a constant pressure of 0.09MPa, and the circulating stainless steel belt is evenly connected to the film. Dry with hot air at 80°C to obtain a roll of 0.03mm thick transparent and dry polysaccharide film. The measured performance indicators are as follows:

[0033] Tensile strength 32Mpa right angle tear strength 38KN / m

[0034] Light transmittance 96% Oxygen transmission rate 1.6cm 3 / m 2 .bar.d

Embodiment 2

[0036] Cut the polysaccharide film prepared in Example 1 into 10mm×10mm unit pieces, place them in a sterile room, and irradiate the film with a 15W ultraviolet lamp at a distance of 20cm for 5 minutes, 10 minutes, 15 minutes, 20 minutes, and 30 minutes respectively. Sterilization, after sterilization, dissolve the diaphragm with a small amount of physiological saline, and then spread it on the plates of complete medium and broth medium respectively and culture it at 30°C for 6 days. The observation results are as follows:

[0037] Irradiation time (minutes) 5 10 15 20 30

[0038] Number of colonies on complete medium 17 3 0 0 0

[0039] Number of colonies on broth medium 13 1 0 0 0

[0040] The polysaccharide membrane can be completely sterilized by ultraviolet irradiation for 15 minutes. For safety reasons, the irradiation time can be extended to 20 minutes

Embodiment 3

[0042] Several polysaccharide membranes prepared in Example 1 were irradiated with ultraviolet rays for 20 minutes for use, and the commercially available nylon composite membranes were cut into a size of 15mm×15mm, and sterilized at 121°C for 30 minutes for use.

[0043] Take the slant strains of Escherichia coli (AS1.751), Corynebacterium Beijing (AS1.299), Bacillus subtilis (AS1.339) and Brevibacterium flavum (AB91058) respectively, smear them on the plate broth medium, and culture them at 32°C , grow into a single colony;

[0044]Smear the yeast (AY91014) on the malt juice agar medium of the plate, culture at 30°C to form a single colony; take out Aureobasidium pullulans AS3. In the sterile room, pick the above-mentioned 6 strains of single colonies from the petri dish (the size of each colony of each strain is basically equal), and spread them on the above-mentioned sterile polysaccharide film irradiated by ultraviolet rays, and then apply a thin layer on the other Glyce...

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Abstract

This invetion relates to Pullulan membrane and its use for culture preservation. The said membrane is prepared by flow spreading process to form thickness 0.02-0.04mm, pull strength 27-32MPa, right tearing strength 31-38KN / m, light transmittance rate 95-98%, and oxygen penetration (2.7-1.6cm3 / m2.bar.d). The method is carried out by: providing glycerin, trehalose as protecting agent and microbe between both of membranes, pressing tightly and sealing with plastic layers to meet conditions for culture preservation, e.g., dryness, oxygen depletion, low temperature, nutrient depletion and protecting agent addition. Those sheets can be formed into albums for preservation, research and easy reproduction by cutting a piece and putting into a culture dish. No expensive apparatus and large space for preservation are needed and it is easily posted.

Description

technical field [0001] The present invention relates to a new microorganism preservation method. Background technique [0002] method advantage shortcoming Slant and Liquid Culture regular transplant preservation Simple, easy to implement, low cost, easy to observe Detection, suitable for all kinds of bacteria The short storage time is 3-6 months, and the bacteria Easy to degrade mineral oil overlay low temperature (refrigerator) Under the given conditions make the strain close to Inactive state, suitable for all kinds of bacteria, The storage time is extended by 1-2 years Occupies a large space and is easily mutated , There are more chances of bacterial infection distilled water Simple and practical, good effect The suitable area is not wide, the storage period is short, the water The quality may affect the preservation of strains Effect Drying method: dry san...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04
Inventor 李世杰
Owner 李世杰
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