Method for preparing L-omithine through immobilized ectocellular enzyme method

An immobilized cell and enzymatic preparation technology, applied in the field of L-ornithine preparation, can solve the problems of low utilization rate of enzyme, reduced product yield, unfavorable continuous production, etc., so as to improve the utilization rate of enzyme and simplify the process. Process, effect of simple product preparation

Inactive Publication Date: 2005-08-31
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The above-mentioned L-ornithinase production process uses free cells to transform the substrate L-arginine, so the transformation solution will contain a small amount of bacterial protein and other impurities, which is not conducive to the separation and purification of the product; and it must be removed after transformation. Bacteria, reducing product yield, resulting in increased production costs
The transformation of free cells is not conducive to industrial continuous production, so the utilization rate of enzymes is not high
[0011] So far, there is no report about the method of immobilized cell enzymatically preparing L-ornithine

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Seed culture and fermentation culture

[0042] A seed medium: 15g glucose, 7.5g beef extract, 15g peptone, 3.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 100mL tomato juice, 900mL water, pH7.5, sterilized at 121°C for 20 minutes.

[0043] A fermentation medium is as follows: glucose 2.0g, yeast extract 0.5g, peptone 0.7g, corn steep liquor 0.8g, potassium dihydrogen phosphate 0.1g, magnesium sulfate 0.06g, L-arginine 0.3g, water 100mL, pH7. 2.

[0044] Melt the freeze-dried Aeromonas intermedius strains with an appropriate amount of sterilized seed medium, and then put it into a 250mL Erlenmeyer flask containing 50mL of liquid seed medium, and cultivate it at a constant temperature of 37°C with a shaker speed of 200r / min. After 24hrs, 10 mL of the above seeds were taken and inserted into fresh seed medium to continue activation.

[0045] Take 4 mL of the above-mentioned activated strains and insert them into 100 mL of fermentation medium, and...

Embodiment 2

[0046] Example 2 Method for preparing L-ornithine by immobilized cell enzymatic method

[0047] As described in Example 1, 1000 mL of Aeromonas intermedia was fermented and cultured, and the thalline was collected by centrifugation at 6000 rpm for 20 min to obtain 12.5 g of wet thalline. Wash into 1200mL fixative (3% sodium alginate solution), and mix well. Then use a syringe to quickly drop the sodium alginate solution containing the bacterium into 3000mL forming agent (2% CaCl 2 solution), the forming agent was removed after curing for 1 hr, and washed 3 times with 5000 mL of physiological saline. The obtained colloidal particles were loaded into a glass column with a diameter of 5 cm and a height of 50 cm. 1000mL of 5% L-arginine solution flowed through the gel column at 20mL / h. After the conversion was complete, the effluent was collected and concentrated to 200mL under reduced pressure, and decolorized with 2g of activated carbon. The decolorized solution was further c...

Embodiment 3

[0048] Example 3 Method for preparing L-ornithine by immobilized cell enzymatic method

[0049] 1000 mL of Enterococcus faecalis was fermented and cultured according to the culture method shown in Example 1, and the bacterial cells were collected by centrifugation at a speed of 6000 rpm for 20 min. Weigh 10.2 g of the bacterial cell wet weight, suspend in 50 mL of physiological saline, and keep warm in a 37° C. water bath. Take 10g of polyvinyl alcohol (PVA), add 50mL of distilled water to soak for 1d, add 0.5g of carrageenan, mix well and put it in a pressure cooker at 110°C for 20min to fully dissolve the PVA, take it out and put it in a water bath at 45°C to keep warm. Then mix the cell suspension with an equal amount of PVA-carrageenan mixture, drop it into the pH6.4 KCl-saturated boric acid solution with a needle, and let it stand at 4°C for 30 hours to prepare the immobilized cell micelles of Enterococcus faecalis . The obtained colloidal particles were loaded into a g...

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Abstract

A process for preparing L-ornithin by imobilized cell enzyme method includes coating the fermented microbe cells by different immobilizing carrier, filling in column, making the L-arginine solution to flow through the column, collecting the flowing-out liquid, concentrating and crystallizing or purifying by cationic exchange column.

Description

1. Technical field [0001] The present invention relates to the method for preparing L-ornithine by using immobilized microbial cells, more specifically the present invention relates to the immobilization of cells containing arginine deiminase and ornithine carbamoyltransferase, through the above-mentioned immobilization The action of an intracellular enzyme converts the substrate L-arginine into L-ornithine. It belongs to the technical field of enzyme engineering. 2. Technical background [0002] The main metabolite of protein in the human body is urea. The liver contains arginase, which can hydrolyze L-arginine to produce L-ornithine and urea. L-Ornithine is also a precursor for the biosynthesis of L-Arginine, which can participate in the composition of proteins. L-ornithine plays an extraordinary role in the human body, and the formation of urea must have the participation of L-ornithine. L-ornithine deficiency can be life-threatening. L-ornithine can be used in the t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/10
Inventor 焦庆才李加友曹瑜刘毅钱绍松陈群
Owner NANJING UNIV
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