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Production of stabilized isotope 15N labelled L-valine

A production process, the technology of valine, applied in the field of producing stable isotope 15N labeled L-valine, can solve the problems such as the decrease of N abundance, achieve the effects of shortening the production cycle, saving recycling costs, and increasing the acid production rate

Inactive Publication Date: 2007-06-20
SHANGHAI RES INST OF CHEM IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Purpose of the present invention is exactly to provide a kind of stable isotope in order to overcome the deficiency that above-mentioned prior art exists 15 The production process of N-labeled L-valine, which uses microbial direct fermentation to produce L-valine- 15 N, which improves growth and metabolism by adding a microvitamin complex designed to address 15 The problem of decreased N abundance, and try to improve 15 N utilization to meet product production requirements

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The slant medium used: glucose 1g / L, beef extract 10g / L, peptone 10g / L, NaCl5g / L, agar 20g / L, fermentation medium: glucose 100g / L, ( 15 NH 4 ) 2 SO 4 15g / L, K 2 HPO 4 1.5g / L, MgSO 4 ·7H 2 O 0.5g / L, FeSO 4 ·7H 2 O 0.02g / L, MnSO 4 ·H 2 O 0.02g / L, biotin 50μg / L, thiamine 100μg / L, corn steep liquor 1g / L, CaCO 3 30g / L.

[0022] Corynebacterium Beijing CGMCC1.586 was inoculated from the preserved slant to the activated slant, cultured in a 30°C constant temperature incubator for 24 hours, washed with a little sterile water, and transferred to a 500mL Erlenmeyer flask containing 20ml of fermentation medium. A total of 10 bottles were fermented on a revolving shaker at 200r / min at 30°C for 60 hours, and the fermentation produced an average of L-valine- 15 N can reach 18g / L.

[0023] The fermentation broth is separated by a 732 cation exchange resin single column, and the dry solid L-valine- 15 The N product was analyzed by mass spectrometry to obtain a product a...

Embodiment 2

[0026]The slant medium used: glucose 1g / L, beef extract 10g / L, peptone 10g / L, NaCl5g / L, agar 20g / L, fermentation medium: glucose 120g / L, ( 15 NH 4 ) 2 SO 4 30g / L, K 2 HPO 4 1g / L, MgSO 4 ·7H 2 O 0.8g / L, FeSO 4 ·7H 2 O 0.05g / L, MnSO 4 ·H 2 O 0.02g / L, biotin 50μg / L, thiamine 100μg / L, cell hydrolyzate 5g / L, CaCO 3 40g / L.

[0027] Inoculate Corynebacterium Beijing CGMCC1.1001 from the preserved slant to the activated slant, cultivate it in a 30°C constant temperature incubator for 18 hours, wash the two slant lawns with a little sterile water, and transfer them to the above-mentioned 500mL Erlenmeyer flask containing 20mL of fermentation medium At 30°C at 200rpm, the fermentation culture was completed on a revolving shaker for 72 hours, and the fermentation produced an average of L-valine- 15 N can reach 25g / L.

[0028] The fermentation broth is separated by a single column of 1500H cation exchange resin, and the dry solid L-valine- 15 The N product was analyzed by ...

Embodiment 3

[0030] Slope medium used: glucose 1g / L, beef extract 10g / L, peptone 10g / L, NaCl5g / L, agar 20g / L, fermentation medium: sucrose 140g / L, ( 15 NH 4 ) 2 SO 4 40g / L, K 2 HPO 4 3g / L, MgSO 4 ·7H 2 O 0.2g / L, FeSO 4 ·7H 2 O 0.02g / L, MnSO 4 ·H 2 O 0.08g / L, biotin 100μg / L, thiamine 50μg / L, corn steep liquor 2g / L, CaCO 3 30g / L.

[0031] Inoculate Brevibacterium flavum ATCC14067 from the preserved slant to the activated slant, and cultivate it in a constant temperature incubator at 30°C for 15 hours, wash the two slant lawns with a little sterile water and transfer them to a 500mL Erlenmeyer flask containing 20mL of fermentation broth, at 30°C 200r / min on the roving shaker for 96 hours of fermentation and cultivation, and the fermentation produces an average of L-valine- 15 N can reach 21g / L.

[0032] Fermentation liquid adopts FMC11Na cation exchange resin single column separation, adopts conventional method to obtain dry solid L-valine- 15 The N product was analyzed by mass...

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Abstract

Production of stable isotope 15N marked L-valine is carried out by selecting proper bacterium, slope preparing, culturing formula by fermentation, fermenting, separating and extracting. It can add micro-vitamin composite, improve growth and metabolism and decrease 15N utilization rate.

Description

technical field [0001] The invention belongs to the technical field of production of stable isotope labeled compounds, in particular to the production of stable isotopes by means of microbial fermentation and biological extraction 15 Process for N labeling L-valine. Background technique [0002] L-valine- 15 Like other labeled amino acids, N can be widely used as a tracer in related research fields such as pharmaceutical industry, food industry and life science. L-valine- 15 The unique tracer function of N can be used to experiment and study the unique physiological function of L-valine in the body. To the production of L-valine, some research groups have done a lot of research work to it at home and abroad, have US 6737255 B2; US4542098; US6214591B; 6(1): P24-27, 1999. However, research on biosynthesis 15 There are not many literatures and patents on the production of N stable isotope-labeled L-valine, such as Acta Microbiology, 2 (2): P75-78, 1994. Produced by direc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/08C12R1/15C12R1/19C12R1/20C12R1/02
Inventor 梅丛笑谭青乔李良君杜晓宁宋明鸣侯静华
Owner SHANGHAI RES INST OF CHEM IND
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