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Method for preparing glutathione by fed-batch fermentation of Candida utilis

A technology of candida utilis and fed-batch fermentation, which is applied in the field of microbial fermentation, can solve the problems of insufficient supply of nutrients, excess nutrition, insufficient supply of yeast nutrients, etc., and achieve the goal of improving production efficiency, reducing fermentation time, and increasing GSH production Effect

Inactive Publication Date: 2012-05-23
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the constant-rate feeding method can meet the nutritional needs of yeast cells in the initial stage of supplementation, but with the continuous increase of the number of yeast cells, constant-speed feeding will inevitably lead to insufficient supply of nutrients; The amount of supplementation in the initial stage of addition is small, but the supplementary amount increases exponentially with time, which will lead to insufficient nutrition supply of yeast in the early stage and excess nutrition in the later stage

Method used

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  • Method for preparing glutathione by fed-batch fermentation of Candida utilis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment one: adopt constant-speed fed-batch fermentation culture under the condition that the total glucose concentration is 150g / L

[0035] (1) strain

[0036] Candida utilis (Candida utilis) SZU 07-01 is preserved by Industrial Microbiology Laboratory of Soochow University.

[0037] (2) culture medium

[0038] Slope and seed medium (g / L): glucose 20, peptone 20, yeast extract 10, pH 6.0;

[0039] Batch fermentation medium (g / L): glucose 30, ammonium sulfate 8, potassium dihydrogen phosphate 3, magnesium sulfate 0.25, pH 5.5;

[0040] Feed medium (g / L): glucose 600, ammonium sulfate 80, potassium dihydrogen phosphate 30, magnesium sulfate 2.5.

[0041] (3) Seed cultivation

[0042] After the slant seeds are activated for 4 to 6 hours, insert them into a 500mL Erlenmeyer flask containing 50mL of seed medium and cultivate them for 18 to 24 hours to obtain the first-grade seeds, and then insert the first-grade seeds into the same medium according to the inoculum am...

Embodiment 2

[0058] Embodiment two: under the condition that total glucose concentration is 150g / L, adopt exponential fed-batch fermentation culture

[0059] With reference to Embodiment 1, the difference is:

[0060] (4) Fermentation tank culture

[0061] Batch fermentation: put the prepared secondary seeds into the fully automatic fermentation tank with 3L fermentation medium according to the inoculum amount of 10%, and cultivate them in batches for 24-30 hours, the fermentation temperature is 28-32°C, and the stirring speed is 300rpm , Ventilation 1vvm.

[0062] Exponential fed-batch fermentation: During 10 to 12 hours of batch fermentation, feeding medium is fed into the fermenter, and the flow acceleration rate F (mL / h) is determined by the exponential equation:

[0063] F = μ ( VX ) 0 Y ...

Embodiment 3

[0069] Embodiment three: under the condition that the total glucose concentration is 150g / L, adopt polynomial fed-batch fermentation culture

[0070] With reference to Embodiment 1, the difference is:

[0071] (4) Fermentation tank culture

[0072] Batch fermentation: put the prepared secondary seeds into the fully automatic fermenter with 3L fermentation medium according to the inoculum amount of 10%, cultivate for 24-30 hours, the fermentation temperature is 28-32°C, the stirring speed is 300rpm, and the ventilation The amount is 1vvm.

[0073] Polynomial fed-batch fermentation: During the 10th to 12th hour of batch fermentation, the feed medium is fed into the fermenter, and the flow acceleration rate F (m L / h) is determined by the polynomial equation:

[0074] F=34.566-4.994*t+1.235*t 2 -0.041*t 3

[0075] The culture conditions are the same as batch culture, and the dissolved oxygen concentration is controlled to not be lower than 35% by manually adjusting the speed ...

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Abstract

The invention belongs to the field of microbial fermentation, and more particularly relates to a method for preparing glutathione by fed-batch fermentation of Candida utilis. The method comprises: taking the Candida utilis SZU 07-01 as original strain, carrying out slant cultivation and seed cultivation on the original strain, and fermenting for at least 42 hour; and fermentation more particularly comprises the steps of: after expansion of cultivation, inoculating seeds into a fermentation medium according to 5-10% of inoculation quantity, and controlling the fermentation temperature within the range of 28-32 DEG C, the stirring rotation speed within the range of 250-300rpm and the ventilation capacity within the range of 0.5-1.5vvm; furthermore, when the fermentation is carried out for 10-12h, feeding the fermentation medium in batch; and manually adjusting the rotation speed and the ventilation capacity, and controlling the concentration of dissolved oxygen to be more than or equal to 35%. By adopting multinomial fed-batch fermentation way, on the basis of meeting the demand for nutrient substance of yeast cells, the method further improves the cell density and GSH yield, obviously reduces the fermentation time and increases the production efficiency.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to a method for preparing glutathione by fed-batch fermentation of Candida utilis. Background technique [0002] Since the earliest patent on the production of GSH by yeast was published in 1938, foreign scholars have carried out a lot of research on the production of GSH. Generally speaking, the production methods of glutathione mainly include solvent extraction, chemical synthesis, and enzymatic conversion. and fermentation. The extraction method is mainly to separate and extract GSH from animal and plant tissues containing GSH by means of extraction and precipitation. Since the raw materials are not easy to obtain and the content of GSH is extremely low, the practical application value of this method is not great. GSH is produced by chemical synthesis, that is, L-glutamic acid, L-cysteine ​​and glycine are condensed into GSH. This method was applied to the prod...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C12R1/72
Inventor 卫功元聂敏王大慧
Owner SUZHOU UNIV
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