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Application of tetra-carbonic acid or salt thereof to preparing 2-keto-L-gulonic acid in tricarboxylic acid cycle

A technology of tricarboxylic acid, gulonic acid, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc.

Inactive Publication Date: 2010-10-06
仪宏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] Before the disclosure of the present invention, there was no literature report on the relationship between 4 kinds of tetracarbonic acid compounds or their salts in the tricarboxylic acid cycle and the growth and acid production of small bacteria

Method used

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  • Application of tetra-carbonic acid or salt thereof to preparing 2-keto-L-gulonic acid in tricarboxylic acid cycle
  • Application of tetra-carbonic acid or salt thereof to preparing 2-keto-L-gulonic acid in tricarboxylic acid cycle
  • Application of tetra-carbonic acid or salt thereof to preparing 2-keto-L-gulonic acid in tricarboxylic acid cycle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Embodiment one: the preparation (preparation of culture medium) of the nutrient-enhanced corn steep liquor product containing succinic acid

[0084] Step 1: Select corn soaking water with stable quality, the dry matter content of which is not less than 9%, add inorganic flocculant, heat to 65°C-85°C and filter,

[0085] Step 2: using succinic acid to acidify the filtrate so that the succinic acid content reaches 5% (w / w) of the soaking water dry matter,

[0086]Step 3: Vacuum concentration until the dry matter reaches 40%, to obtain a concentrated slurry,

[0087] Step 4: Add succinic acid again to the concentrated pulp to obtain a strengthened pulp with a succinic acid content of 10% (mass volume percentage). When the slurry is fermented with 2-KGA, the usage amount is 1% (in terms of dry matter), and the succinic acid in the culture medium reaches 0.1%.

[0088] In this example, the purification of corn steep liquor is combined with the strengthening of tetracarboni...

Embodiment 2

[0090] Embodiment two: Sodium succinate is to the improvement of " two-step fermentation method " technique, bacterial classification adopts large bacterium and small bacterium

[0091] The results of Experiment 2 show that among the four compounds, succinic acid has the most obvious promotion effect on the growth and acid production of small bacteria, so it is used as a representative in the process of fermentation and production of 2-KGA. Since the first step of the "two-step fermentation" process is very mature from sorbitol to sorbose, only the second-step fermentation from sorbose to 2-KGA is used to illustrate the "two-step fermentation" process improvement

[0092] Step 1: The preparation of the culture medium adopts the conventional culture medium preparation method, using sorbose as the substrate:

[0093] 1. Solid medium (mass volume percentage): sorbose 2%, corn steep liquor 1%, yeast powder 1%, urea 1%, potassium dihydrogen phosphate 0.1%, ammonium sulfate 0.1%, s...

Embodiment 3

[0104] Embodiment 3: Using sodium succinate to establish a "pure fermenting process" from sorbose to 2-KGA, the bacterial species adopts small bacteria

[0105] Step 1, the preparation of culture medium, is the same as embodiment 2.

[0106] Step 2, conventional sterilization of the culture medium is the same as in Example 2.

[0107] Step 3: 10-liter fermenter, sterilized by steam for 30 minutes, liquid medium, 121 ° C, sterilized for 25 minutes, sorbose, urea and other medium components were sterilized separately.

[0108] Step 4: Make the small bacteria grown on the solid medium into a bacterial suspension, take 5ml and insert it into the seed medium, the liquid volume is 75ml / 500ml, 28°C-30°C, shaker speed 180-200rpm, cultivate for 18-20h , made into liquid seeds.

[0109] Step 5, liquid seeds, 5% inoculum amount is inserted into a 10L fermenter, 28°C-30°C, 250-300rpm, sodium hydroxide automatically adjusts the pH to 7.2, and cultivates for 48h.

[0110] Step 6: Separat...

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Abstract

The invention relates to an application of tetra-carbonic acid or salts thereof, in particular to application of tetra-carbonic acid or salts thereof in preparation of 2-keto-L-gulonic acid in fermentation engineering and biological technical fields. Meanwhile, the invention further provides a culture medium for preparing 2-keto-L-gulonic acid and a method for preparing 2-keto-L-gulonic acid fromthe tetra-carbonic acid or the salts thereof in the tricarboxylic acid cycle.

Description

technical field [0001] The invention relates to the use of tetracarbonic acid or its salt in the tricarboxylic acid cycle, in particular to the application of the tetracarbonic acid in the tricarboxylic acid cycle or its salt in the preparation of 2-keto-L-gulonic acid in the field of fermentation engineering and biotechnology . Background technique [0002] 2-keto-L-gulonic acid (2-KGA for short) is an important precursor for the preparation of vitamin C (L-ascorbic acid), mainly using the "two-step fermentation" process. [0003] The preparation process of this process needs to go through a two-step fermentation process: the first step of fermentation, with D-sorbitol as the substrate, through the fermentation of acetic acid bacteria Acetobacter sp. to obtain L-sorbose, and the second step of fermentation, with L-sorbitol Sugar is used as a substrate, and 2-KGA is obtained by mixed fermentation of two microorganisms, Ketogulonigenium vulgare (commonly known as small bacte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/60C12R1/01
Inventor 仪宏
Owner 仪宏
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