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Fermentation process for producing stable isotope carbon 13 completely labeled L-shaped leucine

A fermentation process, isotope technology, applied in the direction of fermentation, microorganism-based methods, microorganisms, etc., to achieve the effect of improving utilization, reducing costs, and saving raw materials

Inactive Publication Date: 2012-02-15
SHANGHAI RES INST OF CHEM IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Purpose of the present invention is exactly to provide a kind of can use in order to overcome the defective that above-mentioned prior art exists 13 The C raw material is effectively utilized, and the abundance can be reduced very little, so that high-yield production can be achieved without affecting the abundance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The strain used is Brevibacterium flavum ATCC39106 as the starting bacterium, and the culture medium used includes slant preservation medium, slant activation medium and shake flask fermentation medium. The slant preservation medium and the slant activation medium are the same as those used in conventional fermentation. The formulations of the slant preservation medium, slant activation medium and shake flask fermentation medium used are as follows:

[0023] Incline preservation medium: peptone: 10g / l, beef extract: 3g / l, sodium chloride: 5g / l, agar: 20g / l;

[0024] Incline activation medium: glucose: 3g / l, peptone: 10g / l, beef extract: 10g / l, sodium chloride: 5g / l, agar: 20g / l;

[0025] Fermentation medium: 13 C Glucose: 100g / l, (NH 4 ) 2 SO 4 : 20g / l, MgSO 4 .7H 2 O: 6mg / L, MnSO 4 ·H 2 O: 4mg / L, FeSO 4 ·7H 2 O: 4mg / L, KH 2 PO 4 : 1g / l, VH: 10ug / L, VB1: 400ug / L, pantothenic acid: 100ug / l, nicotinic acid: 100ug / l, corn oatmeal: 0.5g / l, 13 C high-abundance ...

Embodiment 2

[0030] The strain used is Corynebacterium glutamicum ATCC13032 as the starting bacterium, and the culture medium used includes slant preservation medium, slant activation medium and shake flask fermentation medium. The slant preservation medium and the slant activation medium are the same as those used in conventional fermentation. The formulations of the slant preservation medium, slant activation medium and shake flask fermentation medium used are as follows:

[0031] Incline preservation medium: peptone: 10g / l, beef extract: 3g / l, sodium chloride: 5g / l, agar: 20g / l;

[0032] Incline activation medium: glucose: 3g / l, peptone: 10g / l, beef extract: 10g / l, sodium chloride: 5g / l, agar: 20g / l;

[0033] Fermentation medium: 13 C Glucose: 100g / l, (NH 4 ) 2 SO 4 : 35g / l, MgSO 4 .7H 2 O: 6mg / L, MnSO 4 .H 2 O: 2mg / L, FeSO 4 .7H 2 O: 2mg / L, KH 2 PO 4 : 1g / l, VH: 25ug / L, VB6: 100ug / L, pantothenic acid: 300ug / l, niacin: 400ug / l, peptone: 1g / l, 13 C high-abundance cell hydro...

Embodiment 3

[0038] The strain used is Brevibacterium flavum ATCC39103 as the starting bacterium, and the medium used includes slant preservation medium, slant activation medium, seed medium, and fermentation medium. The slant preservation medium and the slant activation medium are the same as those used in conventional fermentation. The formulations of the slant preservation medium, slant activation medium, seed medium, and shake flask fermentation medium used are as follows:

[0039] Incline preservation medium: peptone: 10g / l, beef extract: 3g / l, sodium chloride: 5g / l, agar: 20g / l;

[0040] Incline activation medium: glucose: 3g / l, peptone: 10g / l, beef extract: 10g / l, sodium chloride: 5g / l, agar: 20g / l;

[0041] Seed medium: glucose: 30g / l, urea: 1g / l, ammonium sulfate: 5g / l, KH 2 PO 4 : 0.5g / l, Corn Stew: 20g / l, MgSO 4 .7H 2 O: 5mg / L

[0042] Fermentation medium: 13 C Glucose: 110g / l, (NH 4 ) 2 SO: 40g / l, MgSO 4 .7H 2 O: 6mg / L, MnSO 4 .H 2 O: 2mg / L, FeSO 4 .7H 2 O: 2mg / L...

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Abstract

The invention relates to a fermentation process for producing stable isotope carbon 13 completely labeled L-shaped leucine, which is characterized in that after the bottle shaking fermentation or fermentation tank fermentation is adopted for carrying out fermentation culture on thalli, the isotope C13 completely labeled L-shaped leucine is obtained through separation and extraction, wherein the fermentation tank fermentation is adopted when the product demands are great, and conversely, the bottle shaking fermentation is adopted. Compared with the prior art, the process has the advantages that the acid yield of the obtained <13>C stable isotope labeled L-shaped leucine is correspondingly improved by more than 50 percent than the acid yield adopting a fully synthesized culture medium for fermentation, in addition, the abundance decrease of <13>C can be reduced to a value lower than 0.5 percent, the production cost is reduced, and the fermentation and extraction process is simplified. Raw materials are sufficiently utilized, and the low-abundance and high-abundance <13>C dextrose can be used for meeting the requirements of products with different abundances.

Description

technical field [0001] The invention belongs to the field of stable isotope-labeled compound production and relates to a microbial fermentation production process, in particular to a fermentation process for producing fully-labeled L-type leucine with a stable isotope carbon 13. Background technique [0002] L-leucine-U- 13 C 6 The production of organic synthesis method, precursor fermentation method, enzymatic method and extraction method, direct fermentation method, etc. can be used. The synthesis process is cumbersome, and the result of synthesis is racemized DL-type leucine-U- 13 C 6 , requires optical resolution to obtain the target L-leucine-U- 13 C 6 ,Make 13 The utilization rate of C raw materials is greatly reduced, and the cost rises. Hydrolysis is often used to prepare complex amino acids, and it is difficult to separate all amino acids. Enzymatic Production of L-Leucine-U- 13 C 6 Requires α-oxohexanoic acid, ammonium formate- 13 C is used as a raw mate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/06C12R1/13C12R1/15C12R1/19C12R1/01
Inventor 张亮杜晓宁李良君宋明鸣
Owner SHANGHAI RES INST OF CHEM IND
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