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Method for fermentation shaking flask inoculation

A technology of shake flasks and strains, applied in biochemical equipment and methods, microorganisms, microorganisms, etc., can solve the problems of reduced comparability of shake flask fermentation, inability to ensure accurate and consistent inoculum volume, and bacterial contamination

Inactive Publication Date: 2007-09-26
PETROCHINA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This inoculation method cannot guarantee the exact consistency of the inoculum amount and the consistency of the strain traits of each shake flask due to the differences in the inoculum amount and the character of the thalline, which greatly reduces the comparability of the shake flask fermentation comparative experiment, and each Each shaker needs to use a new inoculation shovel, so a large number of inoculation shovels need to be prepared, and the provenance needs to be repeatedly capped and unplugged during the inoculation process, which is easy to cause bacterial contamination, and the inoculation shovel is used to shovel bacteria from the solid medium When planting, especially when the bacterial cells are thinly or rarely distributed on the solid medium, it is easy to bring the solid medium down

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0008] Detect and compare the applicability of nitrile hydratase-producing bacteria to different yeast extracts. In this experiment, two kinds of yeast extracts were respectively used with 5 parallel 250ml contrast shake flasks. 3 1ml pipettes (1 for each type of yeast paste), add 1ml of bacterial suspension to each shaker flask, after calculation, you need to prepare 1.5ml of bacterial strains, 12ml of deionized water, and then add deionized water to the triangle Add a small amount of 50 clean glass beads to the bottle at the same time, then plug it with gauze, wrap it with kraft paper, put it in a sterilizer, sterilize it at 121°C and 0.11MPa for 30 minutes, and put it away for later use; and prepare 1ml 3 pipettes, insert a cotton column about 1 cm long at 1 cm away from the upper mouth of each pipette, the degree of tightness is suitable for sucking and blowing liquid with the mouth, wrap it in kraft paper, and put it into the sterilizer Sterilize at 121°C and 0.11MPa for ...

Embodiment 2

[0010] Determine the appropriate amount of auxin in the fermentation production of nitrile hydratase-producing bacteria. In this experiment, the auxin content uses four points of 10, 12, 14, and 16ppm, and each point uses 5 parallel 500ml contrast shake flasks, and the shake flasks are fermented The amount of liquid is 50ml, the inoculum amount is 0.5%, 6 pipettes of 2ml are used (one for each value of auxin), and 2ml of bacterial suspension is added to each shaker flask. After calculation, 6ml of strains need to be prepared. Deionized water 48ml, then add deionized water into the triangle flask and add a small amount of cleaned glass beads 50, then plug it with gauze, wrap it with kraft paper, put it in a sterilizing pot at 121°C, 0.11MPa Sterilize for 30 minutes under the same conditions and put it away for later use; and prepare 6 2ml pipettes, and insert a cotton column about 1 cm long at each pipette 1 cm away from the upper mouth, so tight that the liquid can be sucked an...

Embodiment 3

[0012] Determine the appropriate amount of urea added in the fermentation production of nitrile hydratase producing bacteria. In this experiment, the urea content adopts five points of 0.6%, 0.7%, 0.8%, 0.9%, and 1.0%. bottle, the amount of fermented liquid in the shake bottle is 50ml, the inoculum amount is 0.5%, 7 pipettes of 2ml are used (one for each value of urea), and 2ml of bacterial suspension is added to each shake flask, which needs to be prepared after calculation Strain 7.5ml, deionized water 60ml, then add deionized water into the triangular flask and add a small amount of cleaned glass beads 50, then plug it with gauze, wrap it with kraft paper, put it in the sterilizer Sterilize at 121°C and 0.11MPa for 30 minutes and put it away for later use; and prepare 7 2ml pipettes, and insert a cotton column about 1 cm long at each pipette 1 cm away from the upper mouth. It is appropriate to suck and blow the liquid with your mouth, wrap it in kraft paper, put it in a ste...

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PUM

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Abstract

The invention discloses a ferment shaking bottle seeding method to improve comparability of biochemistry shaking bottle ferment comparing experiment, which is characterized by the following: (1) pre-preparing right amount of sterilized water with beaded glass; setting the quantity of sterilizer water at volume of bacterial : density of bacterial suspension; adding into 50-100 particles beaded glass; putting into sterilizing pot; sanitizing at 30 min under 121 deg.c at 0.11MPa; reserving; (2) pre-preparing right amount bacterial; setting the quantity of bacterial at 1.2 X volume of ferment shaking bottle X numbers of ferment shaking bottle X volume density of seeding quantity of ferment shaking bottle; (3) preparing eggplant bottle or test tube to store seed origin; decanting the sterilized water with beaded glass to into eggplant bottle or test tube in the germ-free zone around of alcohol burner; shocking completely to make thallus form bacterial suspending liquid. This invention decreases the repeated time of test, which is fit for various flora of liquid phase ferment culture.

Description

technical field [0001] The invention relates to a fermentation shake flask inoculation method for improving the comparability of biochemical shake flask fermentation comparative experiments. Background technique [0002] In the past, in the comparison experiment of biochemical shake flask fermentation, the seed source was shoveled off the solid-phase medium in the aseptic area around the alcohol lamp in the ultra-clean workbench or in the aseptic room with a pre-sterilized inoculation shovel , an inoculation method for inoculating fermentation shake flasks. This inoculation method cannot guarantee the exact consistency of the inoculum amount and the consistency of the strain traits of each shake flask due to the differences in the inoculum amount and the character of the thalline, which greatly reduces the comparability of the shake flask fermentation comparative experiment, and each Each shaker needs to use a new inoculation shovel, so a large number of inoculation shovels...

Claims

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Application Information

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IPC IPC(8): C12N1/00
Inventor 周云霞吴金海李红梅高俊董文慧刘晶
Owner PETROCHINA CO LTD