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Cultivation method of ketogulonigenium sp

A technology of ketoglobulina and its cultivation method, which is applied in the field of cultivation of ketoglobulina, and can solve problems such as application limitations of enterprises

Inactive Publication Date: 2016-03-02
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing technology uses exogenous additives to try to increase the biomass and acid production of small bacteria, but because the concentration of exogenous additives is directly related to the cost of the enterprise, it has limitations in enterprise application

Method used

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  • Cultivation method of ketogulonigenium sp
  • Cultivation method of ketogulonigenium sp
  • Cultivation method of ketogulonigenium sp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Taking the growth of sorbose-added seed medium as a control, sorbitol, mannitol, fructose, and glucose, which have similar molecular weights to sorbose, were selected as carbon sources that were more beneficial to their growth. Compared with raw materials that are easier to obtain in large-scale industrial applications, sorbitol, as a precursor of sorbose, is cheaper. K.vulgare was cultured for 48 hours to observe the growth and substrate conversion over time, as shown in the figure. The growth of K.vulgare in sorbitol and mannitol was significantly better than that in sorbose, which was 40.72% and 62.97% higher than that in sorbose, which may be related to their transport into the bacteria to participate in multi-step reactions. The curves of the consumption rate of the five substrates over time were obtained by high performance liquid chromatography, such as figure 1 shown. See Table 1 to Table 10 for the original data:

[0041] figure 1 A's original data:

[004...

Embodiment 2

[0065] With the help of next-generation sequencing technology, the whole genome sequencing results of the strain were obtained, and a chromosome and two plasmids were obtained, and the sequences were submitted to the website of the National Center for Biotechnology Information (NCBI) in the United States, and the obtained numbers were CP012908, CP012909 and CP012910 . With the help of KEGGserver online analysis software to obtain the main metabolic pathway for substrate utilization, it was found that the key conversion pathway of carbohydrates in Bacillus ketogenes was seriously lacking. The utilization of different monosaccharides may be mainly realized by its large number of transmembrane dehydrogenase proteins.

Embodiment 3

[0067] In order to further observe the metabolic changes after the utilization of different substrates in cells, the changes of intracellular metabolites in the logarithmic growth phase (12 hours) were obtained by means of metabolomics analysis, and the relative differences of different acidic products were obtained. Among them, sorbitol and 2-keto-L-gulonic acid were detected when sorbitol was used as the substrate. It shows that the strain also has the mechanism of metabolizing sorbitol to produce acid. Mannose was detected when mannitol was used as the substrate. In addition, under the five substrates, the products 2-keto-D-gluconic acid, L-gluconic acid, glucaric acid, and D-gluconic acid have obvious differences, which are related to different metabolic methods. The raw data are shown in Tables 11-15 (relative quantity, no unit):

[0068] Table 11 The content of different products in the cells under the addition of sorbose in the logarithmic phase

[0069]

[0070] ...

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Abstract

The invention relates to the field of microorganisms, in particular to a cultivation method of ketogulonigenium sp. A new technological process of vitamin C precursor 2-ketone-L-gulonic acid is established, in the process, a brand-new ketogulonigenium sp seed cultivation mode is used, and sorbitol and / or mannitol serve / serves as a carbon source in replacement of sorbose. Under the same cultivation time and cultivation condition, the mycelia content is increased by 1.6 times or more through seed cultivation of sorbitol / mannitol than a sorbose cultivation mode, and high-quality fermentation strains are provided for follow-up fermentation. As for the aspect of the same mycelia content, the highest biomass obtained after 25 hours of cultivation in sorbose can be obtained after 5 hours of cultivation in sorbitol and mannitol. Accordingly, the seed cultivation time is greatly shortened by at least 10 hours.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a method for cultivating ketoglobulina. Background technique [0002] Vitamin C is a water-soluble vitamin necessary to maintain the growth and metabolism of the body. It is currently widely used in the fields of medicine, food, feed and cosmetics, and the market is stable. At present, the mainstream production method of vitamin C is the two-step fermentation method, which mainly refers to the process of starting from D-sorbitol and converting it into vitamin C precursor—2-keto-L-gulonic acid through two-step fermentation. In the first step, use Gluconobacter oxydans to convert D-sorbitol into L-sorbose, and in the second step, use a mixed system of ketobacterium and its associated bacteria (mainly Bacillus) to convert L-sorbose For 2-keto-L-gulonic acid. Because the strain grows slowly when cultured alone, it affects its industrial application in amplification culture and strain ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/60C12R1/01
CPCC12N1/20C12P7/60
Inventor 元英进贾楠丁明珠潘才惠
Owner TIANJIN UNIV
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