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A kind of fermentation method that improves the acid production of gulong acid

A fermentation method and technology of cologne acid, applied in the field of microbial fermentation, can solve the problems of complexity, control, shortening fermentation cycle, etc., and achieve the effects of increasing content, improving fermentation production efficiency, and increasing acid production concentration

Active Publication Date: 2017-03-01
ANHUI BBCA FERMENTATION TECH ENG RES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The two-step fermentation method uses a mixed-bacteria fermentation system composed of Gluconobacter oxidans and Bacillus megaterium, which makes the control of the fermentation process more complicated. The ratio of the two bacteria has a significant impact on the production of 2-KLG, and it is difficult to adjust the ratio of the two bacteria in actual production. control
[0004] After searching, it is found that the publication number is CN103627775A, and the title of the invention is a method for improving the fermentation production efficiency of 2-keto-L-gulonic acid. It discloses a method for improving the fermentation production efficiency. The fermentation cycle was completed, but there was no significant increase in acid production

Method used

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Examples

Experimental program
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Effect test

preparation example Construction

[0027] Bacterial liquid preparation adopts conventional preparation methods, which can be but not limited to the following preparation methods:

[0028] Cryopreservation tube--shake flask activation for 24 hours--separate large and small bacteria (cultivate small bacteria at 28°C for 2 days, large bacteria for 2-3 days)--choose full colonies, neat edges, and large single colonies of small bacteria Ring-coated test tubes (incubated at 28°C for one day)--take large bacteria, select large colonies with full colonies, neat edges, and a single colony of moderate-sized large bacteria on the test tubes that have been covered with small bacteria (incubate at 28°C for 2 days)--use Wash the bacterial lawn on the test tube with sterile water and inoculate the bottle (28°C, 200r / min, culture for 22h).

[0029] Solid medium is used for strain separation and test tube preparation, and liquid seed medium is used for cryopreservation tube activation and seed expansion cultivation. Solid mediu...

Embodiment 1

[0034] The initial medium for fermentation includes sorbose 2mg / 100ml, corn steep liquor 1.2mg / 100ml, KH 2 PO 4 0.05mg / 100ml, MgSO 4 .7H 2 O0.02mg / 100ml, urea 0.2mg / 100ml, the initial pH is 6.7, wherein feed concentration is the sorbose of 200mg / ml, and the initial sorbose is sterilized separately from other mediums.

[0035] The fermentation method that present embodiment improves the acid production of gulong acid comprises the following steps:

[0036] (1) Insert 3L of the cultured mixed bacteria solution of Gluconobacter oxydans and Bacillus megaterium into 10L containing 2mg / 100ml sorbose, 1.2mg / 100ml corn steep liquor, 0.2mg / 100ml urea, 0.02mg / 100ml MgSO 4 ·7H 2 O, 0.05mg / 100ml of KH 2 PO 4 In the fermentation medium, the fermentation container adopts 30L automatic fermenter.

[0037](2) The first fermentation stage: the pH value of the fermented liquid is controlled to be 6.7, the stirring speed is 350rpm, the aeration ratio is 1:0.6vvm, and 300mg / ml glucose sol...

Embodiment 2

[0045] The preparation method of this example is the same as that of Example 1, except that the initial fermentation medium is the only difference. The fermentation initial culture medium of the present invention comprises 2.5mg / 100ml sorbose, 0.9 / 100ml corn steep liquor, 0.1 / 100ml urea, 0.04mg / 100ml MgSO by mass volume ratio 4 ·7H 2 O, 0.08mg / 100ml of KH 2 PO 4 , the pH value of the initial fermentation medium is 7.0, and the concentration of 250 mg / ml sorbose is added therein, and the initial sorbose is sterilized separately from other mediums.

[0046] In this implementation, the 2-keto-L-gulonic acid content at the end point of the fermentation broth was 93.2 mg / ml, the fermentation period was 47 hours, and the acid production rate was 1.98 g / l / h.

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Abstract

The invention relates to the technical field of microbial fermentation, and especially relates to a fermentation method for increasing gulonic acid to prepare acid. The fermentation method is characterized in that a mixing mycelium of bacillus megaterium and gluconobacter oxydans is taken as bacterial strain, sorbose is taken as a raw material for aerobic fermentation, glucose is added at an early stage, peptone beef is added at an intermediate and later stage; addition of glucose at the early stage enables rapid growth of bacillus megaterium, and addition of peptone beef at the intermediate and later stage can solve the problems of slow growth of small bacterium, short acid production phase and slow acid production. The method is capable of increasing the content of fermentation terminal point gulonic acid, increasing acid production concentration, increasing the fermentation production efficiency, and successfully using on a 500L of fermentation cylinder.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a fermentation method for improving the acid production of gulonic acid. Background technique [0002] At present, domestic production of gulonic acid mostly adopts a two-step fermentation method. The two-step fermentation method usually uses a mixed strain of bacteria for fermentation. Bacillus (commonly known as large bacteria), among which the small bacteria are acid-producing bacteria, which secrete L-sorbosone oxidase in the cell, which can oxidize sorbose into 2-keto-L-gulonic acid (2-KLG), but grow alone Slow, low acid production capacity; large bacteria cannot produce acid, but can be used as companion bacteria for small bacteria, play an auxiliary role in fermentation, and promote the growth and acid production of small bacteria. Studies have confirmed that both the intracellular fluid and extracellular fluid of large bacteria can promote the growth of sm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/60C12R1/11C12R1/01
Inventor 李荣杰尚海涛杨为华邓远德徐斌穆晓玲王芳
Owner ANHUI BBCA FERMENTATION TECH ENG RES
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