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Preparation and regeneration method of zymononas mobilis protoplast

A technology of Zymomonas and protoplasts, which is applied in the field of preparation and regeneration of Zymomonas mobilis protoplasts, can solve the problems of narrow carbon source utilization range and harsh nutritional conditions, and achieve good protoplast activity and high preparation rate , the effect of simple operation

Inactive Publication Date: 2015-03-04
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the defect of Z.mobilis fermentation to produce ethanol is that its carbon source utilization range is very narrow, and only glucose, fructose and sucrose can be used; the nutritional conditions are harsh, and a variety of nutrients need to be added

Method used

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  • Preparation and regeneration method of zymononas mobilis protoplast
  • Preparation and regeneration method of zymononas mobilis protoplast

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Step 1, prepare Z.mobilis slant medium:

[0041] Weigh 10.00g of glucose, 0.50g of yeast extract, 0.10g of potassium dihydrogen phosphate, 0.05g of magnesium sulfate, 0.10g of ammonium sulfate, 2.00g of agar powder, add distilled water to 100mL, and autoclave at 121°C for 30min. The volume of the culture medium in the test tube is 15.00mL, and placed on an inclined plane to form a culture medium for later use;

[0042] Step 2, activate the bacteria:

[0043]Take the ampoule tube preserved with the dry powder of Z.mobilis bacteria. The quality of Z.mobilis dry powder in the ampoule tube is 1.00g. Wipe the ampoule tube with absorbent cotton soaked in 75% (v / v) alcohol, and heat the top of the ampoule tube to red with flame , drop 1.00mL of sterile water to the heated top to make it rupture, knock off the top of the broken ampoule with a file or tweezers, draw 0.50mL of sterile water with a sterile straw, drop it into the ampoule, shake gently to make the bacteria The ce...

Embodiment 2

[0060] Step 1, prepare Z.mobilis slant medium:

[0061] Weigh 10.00g of glucose, 0.50g of yeast extract, 0.10g of potassium dihydrogen phosphate, 0.05g of magnesium sulfate, 0.10g of ammonium sulfate, 2.00g of agar powder, add distilled water to 100.00mL, and autoclave at 121°C for 30min. , the volume of the culture medium in the test tube is 15.00mL, and placed on an inclined plane to form a culture medium for later use;

[0062] Step 2, activate the bacteria:

[0063] Take the ampoule tube preserved with the dry powder of Z.mobilis bacteria. The quality of Z.mobilis dry powder in the ampoule tube is 1.00g. Wipe the ampoule tube with absorbent cotton soaked in 75% (v / v) alcohol, and heat the top of the ampoule tube to red with flame , drop 1mL of sterile water to the heated top to make it rupture, knock off the top of the broken ampoule tube with a file or tweezers, draw 0.50mL of sterile water with a sterile straw, drop it into the ampoule tube, and shake gently to make the...

Embodiment 3

[0080] Step 1, prepare Z.mobilis slant medium:

[0081] Weigh 10.00g of glucose, 0.50g of yeast extract, 0.10g of potassium dihydrogen phosphate, 0.05g of magnesium sulfate, 0.10g of ammonium sulfate, 2.00g of agar powder, add distilled water to 100.00mL, and autoclave at 121°C for 30min. , the volume of the culture medium in the test tube is 15.00mL, and the culture medium is placed on an inclined plane for later use;

[0082] Step 2, activate the bacteria:

[0083] Take the ampoule tube preserved with the dry powder of Z.mobilis bacteria. The quality of Z.mobilis dry powder in the ampoule tube is 1.00g. Wipe the ampoule tube with absorbent cotton soaked in 75% (v / v) alcohol, and heat the top of the ampoule tube to red with flame , drop 1.00mL of sterile water to the heated top to make it rupture, knock off the top of the broken ampoule with a file or tweezers, draw 0.50mL of sterile water with a sterile straw, drop it into the ampoule, shake gently to make the bacteria The...

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Abstract

The invention discloses a preparation and regeneration method of zymononas mobilis protoplast. The preparation and regeneration method comprises the following steps: preparing a bacterium inclined cultivation medium, activating a thallus, preparing a seed culture liquid, performing enzymolysis on cell walls, preparing protoplast suspension, and regenerating protoplast. By adopting the preparation and regeneration method of the zymononas mobilis protoplast, the preparation rate of the protoplast of the zymononas mobilis can be up to 90.0% at most, the regeneration rate can be up to 22.4%, the operation process is simple, high in repeatability and high in preparation rate, the activity of the protoplast is relatively well maintained, and the later preparation and the screening of fusant are facilitated.

Description

technical field [0001] The invention relates to the technical field of bacterial protoplast preparation and regeneration, in particular to a preparation and regeneration method of Zymomonas mobilis protoplast. Background technique [0002] Zymomonas mobilis (Zymomonas mobilis) is a species of Zymomonas genus, has a strong motility, is a Gram-negative, facultative anaerobic bacteria. Zymomonas mobilis has been used in the fermentation production of fruit wine, and its fermentation ability is outstanding. Compared with yeast, the bacterium has many advantages, such as high sugar absorption efficiency, high ethanol yield, less by-products, strong ethanol tolerance, no need to control oxygen addition during fermentation, and easy genetic manipulation. However, the defect of Z.mobilis fermentation to produce ethanol is that its carbon source utilization range is very narrow, only glucose, fructose and sucrose can be utilized; the nutritional conditions are harsh, and a variety o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
CPCC12N1/20
Inventor 龚国利史政豪
Owner SHAANXI UNIV OF SCI & TECH
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