Nutrition-enhanced culture medium for preparing 2-KGA through fermentation and method thereof for preparing 2-KGA

A 2-KGA, nutrient-enhanced technology, applied in the field of a culture medium in the fields of fermentation engineering and biotechnology

Inactive Publication Date: 2010-09-29
仪宏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Before the disclosure of the present invention, there was no close relationship between the above-mentioned compounds and the growth and acid production of small bacteria reported in any literature

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment one: Sodium succinate is to the improvement of " two-step fermentation method " technique

[0046] Bacteria used: small bacteria and large bacteria

[0047] Step 1 medium preparation and sterilization: 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%, amber Sodium phosphate 0.1%, agar 1.8%, pH7.0. Liquid seed medium (mass volume percentage): sorbose 5%, corn steep liquor 1.2%, urea 1.2%, sodium succinate 0.15%, potassium dihydrogen phosphate 0.1%, ammonium sulfate 0.1%, calcium carbonate 0.3%, trace elements and Each vitamin 1ml / L, pH7.0. Liquid fermentation medium (mass volume percentage): sorbose 8%, corn steep liquor 1%, urea 1%, sodium succinate 0.15%, potassium dihydrogen phosphate 0.1%, ammonium sulfate 0.1%, calcium carbonate 0.3%, trace elements and Each vitamin 1ml / L, pH7.0.

[0048] The medium is routinely sterilized at 121°C for 25 minut...

Embodiment 2

[0054] Embodiment 2: Adopt sodium succinate to establish "pure breed fermentation new technology" from sorbose to 2-KGA

[0055] Bacteria used: small bacteria

[0056] Step 1 Culture medium preparation and sterilization: same as Example 1.

[0057] Step 2 Strain preparation and fermentation: Inoculate small bacteria on solid medium at 28°C, culture for 36 hours, make bacterial suspension, insert into liquid seed medium; liquid volume 75ml / 500ml, 28°C-30°C, shaker Rotate at 180-200rpm, cultivate for 18-20h, put 5% inoculum into 10L fermenter, 28°C-30°C, 250-300rpm, sodium hydroxide automatically adjusts pH7.2.

[0058] Step 3 Extracting and refining 2-KGA: proceed according to conventional methods, including flocculation, filtration, ion exchange, decolorization, concentration, and crystallization.

[0059] Detection method: with embodiment one.

[0060] Results: In the absence of Bacillus, sodium succinate can also greatly increase the acid production rate of small bacteria...

Embodiment 3

[0061] Embodiment three: Adopt sodium succinate to establish the new process of "single-bacteria one-step fermentation" from sorbitol to 2-KGA

[0062] Bacteria used: small bacteria

[0063] Step 1 Medium preparation and sterilization: use sorbitol instead of sorbitol, and the others are the same as in Example 1.

[0064] Step 2 Strain preparation and fermentation: inoculate small bacteria on solid medium at 28°C, culture for 36 hours, and make bacterial suspension; insert liquid seed medium, liquid volume 75ml / 500ml, 28°C-30°C, shaker Rotate at 180-200rpm, cultivate for 18-20h; insert 5% (v / v) of the shaker flask seeds into a 500ml pH self-controlling shaker flask, 30°C, 200rpm, and sodium hydroxide automatically adjusts the pH to 7.2.

[0065] Step 3 Extracting and refining 2-KGA: proceed according to conventional methods, including flocculation, filtration, ion exchange, decolorization, concentration, and crystallization.

[0066] Detection method: with embodiment one.

...

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Abstract

The invention relates to a culture medium in the technical fields of fermentation engineering and biology, in particular to a nutrition-enhanced culture medium for preparing 2-KGA through fermentation, and the nutrition-enhanced culture medium comprises a conventional culture medium and a nutrition enhancer, wherein sorbitol or sorbose is used as a substrate of the conventional culture medium. Meanwhile, the invention also relates to a method for preparing the 2-KGA by applying the enhanced culture medium. The nutrition-enhanced culture medium can promote the growth of small bacteria, enablesthe small bacteria to be disengaged from the dependence on associated bacteria, establishes independent growth and provides a foundation for the improvement of a 2-KGA fermentation process.

Description

[0001] Field: [0002] The invention relates to a culture medium in the fields of fermentation engineering and biotechnology, in particular to a nutrition-intensified culture medium for fermenting and preparing 2-KGA, and also relates to a method for preparing 2-KGA by using the strengthened culture medium. Background technique [0003] 2-keto-L-gulonic acid (2-KGA for short) is an important precursor for the preparation of vitamin C (L-ascorbic acid). At present, the industry mainly adopts the "two-step fermentation" process. The process requires a two-step fermentation process: the first step of fermentation, with D-sorbitol as the substrate, and the fermentation of Acetobacter sp. to obtain L-sorbose, and the second step of fermentation, with L-sorbose as the substrate 2-KGA is obtained through the mixed fermentation of two microorganisms, Ketogulonigenium vulgare (commonly known as small bacteria) and Bacillussp (commonly known as large bacteria). The first step of ferme...

Claims

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Application Information

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IPC IPC(8): C12P39/00C12P7/60C12R1/01C12R1/02C12R1/07
Inventor 仪宏
Owner 仪宏
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