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Preservation method for vitamin C mixed bacteria

A technology of vitamin and mixed bacteria, which is applied in the field of preservation of vitamin C mixed bacteria, can solve the problems that the stability cannot be effectively controlled and guaranteed, the proportion of bacteria and bacteria is out of balance, and the hidden dangers of product quality can be prevented, so as to prevent bacterial contamination or Seed solution quality degradation, fermentation process stability, and product quality assurance effects

Inactive Publication Date: 2014-01-08
NINGXIA QIYUAN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to frequent subculture and activation, the bacteria are prone to imbalance in the proportion of large and small bacteria, and the ability to produce acid is reduced. Even because of repeated transplantation and subculture, it is easy to cause large-scale bacterial contamination, so that the stability of production cannot be effectively controlled and guaranteed. , because of the fluctuation of fermentation production level, it brings technical difficulties to the post-process production, and also buryes hidden dangers for product quality problems

Method used

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  • Preservation method for vitamin C mixed bacteria
  • Preservation method for vitamin C mixed bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Preparation and preservation of original bacterial strain cryopreservation tubes

[0023] 1 Take out 1ml of the seed liquid of the original bacterial strain that has been screened by shaking flasks and has excellent production performance and put it into 9ml sterile water with a concentration of 10 -1 , then from 10 -1 Inhale 1ml, put in another 9ml sterile water with a concentration of 10 -2 , and so on, diluted to 10 -8 , then from 10 -8 Aspirate 0.1ml into the separation plate, then spread it evenly with a triangular grill, and culture it in a constant temperature room at 29.5±1°C for 60-72 hours.

[0024] 2. Preparation of inclined plane: From the well-grown separation plate, pick well-grown, translucent, evenly distributed, and free of miscellaneous bacteria, and put them into sterile water to make the water of the small bacteria milky white. Use an inoculation loop or an inoculation stick to inoculate to On the inclined surface of the eggplant-shape...

Embodiment 2

[0027] Example 2: Preparation and preservation of cryopreservation tubes for bacterial strains used in production

[0028] 1 Take 0.5ml of the original bacterial strain cryopreservation tube, inoculate it into the seed medium (20ml), and cultivate it on a shaker at 29.5±1°C (rotating speed: 200 rpm) for 24 hours.

[0029] 2 Separate the cultured seed solution on the plate, spread it evenly, and cultivate it at 29.5±1°C for 2-3 days.

[0030] 3. From the cultured separation plate, select well-grown, translucent, evenly distributed, and free of miscellaneous bacteria, and put them into sterile water to make the water milky white, and inoculate them into a 250ml eggplant-shaped bottle with an inoculation loop or an inoculation stick. On the slope, wrap the mouth of the bottle with two layers of gauze, put it in a constant temperature room at 29.5±1°C for 22 to 24 hours, and observe that there are no bacteria growing, then transfer it to the inoculation room again. For moderately...

Embodiment 3

[0032] Embodiment 3: for the preparation of production seed solution

[0033] 1. Activation of the first generation: Take a cryopreservation tube for the strains used for production, and use a 1ml straw to transfer all the frozen liquid into a triangular flask containing 20ml of seed medium at 29.5±1°C, cultivate for 20-24 hours, and become the first generation seed liquid .

[0034] 2. Activate the second generation: absorb 5ml of the first generation seed solution and inoculate it into another seed medium (20ml) Erlenmeyer flask, at 29.5±1°C, shake and cultivate for 20±4 hours, which is the second generation seed solution.

[0035] 3. Activation of the third generation: inoculate 5ml of the second-generation seed liquid into another Erlenmeyer flask of seed medium (20ml), culture with shaking at 29.5±1°C for 20±4 hours. At this time, when it is detected that the acid production is above 5 mg / ml and there is no miscellaneous bacteria, it can be used for production.

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Abstract

The invention relates to a preservation method for vitamin C mixed bacteria. The method comprises the following steps: inoculating the vitamin C mixed bacteria cultured by matching big bacteria with small bacteria on an inclined plane, preparing a suspension solution by using glycerol in the volume concentration of 50%, further packaging in a sterile freezing tube, and placing at the temperature of -70+ / -2 DEG C for preservation. As for a VC two-step bacterial strain preserved by adopting the method, the death rate of the strain is below 1%, the mutation rate is zero, the properties and the stability of the strain can be kept for above five years, the time consumption is short, the operation method is simple and easy, the two-step bacterial fermentation level after activation is basically kept at the level of 115-125mg / ml, the two-step conversion rate is above 94%, the production is stabilized, and the risk caused by frequent passages is avoided.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, in particular to a method for preserving vitamin C mixed bacteria. Background technique [0002] Vitamin C (Vitanun C, Vc) is an essential vitamin for human nutrition. It is not only used as an important medical product to treat various diseases, but also widely used in food, feed and cosmetics. With the increase of the application range of vitamin C, the market demand is increasing day by day, so that people continue to study and improve the production technology of VC. The fermentative production of vitamin C in the current technology usually adopts a two-step method, wherein the synthesis of gulonic acid: adopt Bacillus megaterium (commonly known as large bacteria, large bacteria are Gram-positive bacillus, which can dye phages and are easier to differentiate, ) and Gluconobacter oxidans (commonly known as small bacteria, small bacteria are Gram-negative non-bacillus, the bact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04
Inventor 李学兵于新燕胡继红陈惠琴
Owner NINGXIA QIYUAN PHARMA
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