Method for improving gluconobacter oxydans to produce 2-keto-L-gulonic acid

A technology of Gluconobacter oxydans and Gulonic acid, which is applied in the field of microbial fermentation, can solve the problems of increased cost, slow growth, and increased cost, and achieve the effects of reducing dosage, increasing growth speed, and reducing costs

Active Publication Date: 2009-12-16
TIANJIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The mixed strains used in the mixed fermentation are Bacillus megaterium and Gluconobacter oxidans mixed bacteria fermentation, wherein Gluconobacter oxidans is an acid-producing bacterium, and Bacillus megaterium is an associated bacterium. Glycobacterium, the growth is extremely slow, and the acid production efficiency is low. When Bacillus megaterium is added, the growth rate of Gluconobacter oxydans can be increased, and...

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  • Method for improving gluconobacter oxydans to produce 2-keto-L-gulonic acid
  • Method for improving gluconobacter oxydans to produce 2-keto-L-gulonic acid
  • Method for improving gluconobacter oxydans to produce 2-keto-L-gulonic acid

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Embodiment 1

[0031] The method for improving the production of 2-keto-L-gulonic acid by Gluconobacter oxydans comprises the following steps:

[0032] (1) Seed cultivation:

[0033] Weigh: L-sorbose 20g, corn steep liquor 3g, beef extract 3g, yeast extract powder 3g, urea 1g, peptone 10g, KH 2 PO 4 1g, MgSO 4 0.2g, CaCO 3 Add 1 g of water to 1 L, adjust the pH to 6.8, and sterilize at 121°C for 20 minutes to make a seed medium;

[0034] Inoculate Gluconobacter oxydans (Gluconobacter oxydans) CGMCC NO.1.110 on the slant into the seed medium, shake at 30°C and 220r / min on a shaker, and cultivate for 36 hours to prepare a seed culture solution;

[0035] (2) Fermentation:

[0036] Weighing: L-sorbose 80g, corn steep liquor 20g, urea 12g, KH 2 PO 4 1g, MgSO 4 0.5g, CaCO 3 Add 5 g of water to 1 L, adjust the pH to 7.0, and sterilize at 121°C for 20 minutes to make a fermentation medium;

[0037] The seed culture solution was inserted into the fermentation medium at a ratio of 10% by vol...

Embodiment 2

[0040] The method for improving the production of 2-keto-L-gulonic acid by Gluconobacter oxydans comprises the following steps:

[0041] (1) Seed cultivation:

[0042] Weigh: L-sorbose 10g, corn steep liquor 2g, beef extract 2g, yeast extract powder 2g, urea 0.5g, peptone 2g, KH 2 PO 4 0.5g, MgSO 4 0.1g, CaCO 3 Add 0.5 g of water to 1 L, adjust the pH to 7.0, and sterilize at 121°C for 20 minutes to make a seed medium;

[0043] Inoculate Gluconobacter oxydans (Gluconobacter oxydans) CGMCC NO.1.110 on the slant into the seed medium, shake at 28°C and 160r / min on a shaker, and cultivate for 48 hours to prepare a seed culture solution;

[0044] (2) Fermentation:

[0045] Weighing: L-sorbose 40g, corn steep liquor 10g, urea 10g, KH 2 PO 4 0.5g, MgSO 4 0.2g, CaCO 3 Add 1 g of water to 1 L, adjust the pH to 7.5, and sterilize at 121°C for 20 minutes to make a fermentation medium;

[0046] The seed culture solution was inserted into the fermentation medium at a ratio of 5% ...

Embodiment 3

[0049] The method for improving the production of 2-keto-L-gulonic acid by Gluconobacter oxydans comprises the following steps:

[0050] (1) Seed cultivation:

[0051] Weighing: 50g of L-sorbose, 10g of corn steep liquor, 10g of beef extract, 10g of yeast extract powder, 5g of urea, 10g of peptone, KH 2 PO 4 5g, MgSO 4 0.7g, CaCO 3 Add 5 g of water to 1 L, adjust the pH to 6.5, and sterilize at 121°C for 20 minutes to make a seed medium;

[0052] Inoculate Gluconobacter oxydans (Gluconobacter oxydans) CGMCC NO.1.110 on the slant into the seed medium, shake at 35°C and 250r / min on a shaker, and cultivate for 24 hours to prepare a seed culture solution;

[0053] (2) Fermentation:

[0054] Weighing: L-sorbose 120g, corn steep liquor 50g, urea 25g, KH 2 PO 4 3g, MgSO 4 1.2g, CaCO 3 Add 10g of water to 1L, adjust the pH to 6.5, and sterilize at 121°C for 20 minutes to make a fermentation medium;

[0055] The seed culture solution was inserted into the fermentation medium ...

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Abstract

The invention discloses a method for improving gluconobacter oxydans to produce 2-keto-L-gulonic acid, which comprises the following steps: (1) seed culture, namely inoculating bevel gluconobacter oxydans into a seed culture medium to form seed culture solution; and (2) fermentation, namely inoculating the seed culture solution into a fermentation culture medium, oscillating the mixture in a shaking bed, carrying out fermentation culture for 48 to 96 hours, and adding a sulfhydryl compound into the mixture at any moment of the fermentation culture in a range of 0 to 36 hours to make the final concentration of sulfhydryl between 0.1 and 30mM. The method can improve the growth speed of the gluconobacter oxydans and the efficiency for producing the 2-keto-L-gulonic acid, and achieve the aim of partially replacing the companion effect of companion fungus so as to reduce the using amount of bacillus megaterium of the companion fungus, reduce the cost of the culture medium and reduce the pollution.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, in particular to a method for improving the production of 2-keto-L-gulonic acid by Gluconobacter oxydans. Background technique [0002] Vitamin C is a trace water-soluble vitamin necessary for the nutrition and growth of the body, and plays an important role in anti-oxidation and maintaining metabolic balance. Vitamin C can be used as medicine, health products, food additives and cosmetic nutrients, its application scope is expanding and the market is stable. [0003] At present, the method of producing vitamin C in my country is a "two-step fermentation method". The first step of fermentation uses Acetobacter black to convert sorbitol into L-sorbose. Precursor 2-keto-L-gulonic acid. The mixed strain used in the mixed fermentation is Bacillus megaterium and Gluconobacter oxidans mixed bacteria fermentation, wherein, Gluconobacter oxidans is an acid-producing bacterium, Bacillus megaterium i...

Claims

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Application Information

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IPC IPC(8): C12P7/60C12R1/01
Inventor 元英进仪宏王丽丽周剑马倩薛佳
Owner TIANJIN UNIV
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