A kind of production and extraction process of l-isoleucine

A technology of isoleucine and extraction process, applied in the directions of biochemical equipment and methods, microorganisms, chemical instruments and methods, etc., can solve the problems of production waste, consumption of reagents, increase process complexity and workload, etc., to improve fermentation Yield, product cost saving, small footprint effect

Active Publication Date: 2021-09-28
XINJIANG FUFENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First: The traditional extraction process mainly uses separation, which consumes a lot of reagents, and requires additional procedures to remove these reagents; this not only increases the complexity and workload of the process, but also causes unnecessary production waste

Method used

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  • A kind of production and extraction process of l-isoleucine
  • A kind of production and extraction process of l-isoleucine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A production and extraction process of L-isoleucine, which comprises the following steps:

[0037] Step 1): Cultivate Corynebacterium glutamicum (CGMCC NO.12153, donated by Tianjin University of Science and Technology) to a concentration of 5×10 8 The seed solution of CFU / mL was then inoculated into the fermentation medium according to 10% inoculation amount, and the fermentation culture was carried out for 72 hours. Isoleucine fermentation broth; the components of the fermentation medium are: glucose 120g / L, (NH 4 ) 2 SO 4 20g / L, FeSO 4 ·7H 2 O 0.015g / L, MgSO 4 ·7H 2 O0.4g / L, MnSO 4 ·H 2 O 0.015g / L, KH 2 PO 4 2.2 g / L, K 2 HPO 4 1.0 g / L, VH 80 μg / L, VB 1 2.5mg / L, Met30mg / L, Glu 20mg / L, Inositol 20mg / L;

[0038]Step 2): pass the L-isoleucine fermentation broth through a ceramic membrane (20nm) to remove bacterial protein; membrane filtration operating temperature: 45°C, working pressure difference of 1.2bar, to obtain the retentate and filtered supernatant;...

Embodiment 2

[0046] A production and extraction process of L-isoleucine, which comprises the following steps:

[0047] Step 1): Cultivate Corynebacterium glutamicum (CGMCC NO.12153, donated by Tianjin University of Science and Technology) to a concentration of 5×10 8 The seed liquid of CFU / mL was then inoculated into the fermentation medium according to 10% inoculation amount, and the fermentation culture was carried out for 75 hours. Isoleucine fermentation broth; the components of the fermentation medium are: glucose 160g / L, (NH 4 ) 2 SO 4 30g / L, FeSO 4 ·7H 2 O 0.015g / L, MgSO 4 ·7H 2 O0.4g / L, MnSO 4 ·H 2 O 0.015g / L, KH 2 PO 4 2.2 g / L, K 2 HPO 4 1.0 g / L, VH 80 μg / L, VB 1 2.5mg / L, Met30mg / L, Glu 20mg / L, Inositol 20mg / L;

[0048] Step 2): pass the L-isoleucine fermentation broth through a ceramic membrane (50nm) to remove bacterial protein; membrane filtration operating temperature: 50°C, working pressure difference of 1.5bar, to obtain the retentate and filtered supernatant; ...

Embodiment 3

[0056] Taking 200L fermentation medium as an example, a total of 6.53kg of L-isoleucine products were obtained. The appearance of the product was white and translucent. The purity is 99.4%.

[0057] The present invention also detects the influence of culture medium and culture time on the concentration of L-isoleucine in the fermentation broth.

[0058] With embodiment 1 as experimental group, the Glu concentration in the culture medium is set to be 0, 10, 20, 30, 40, 50 (mg / L), such as figure 1 As shown, with the increase of Glu concentration, the output of L-isoleucine increased significantly, when the Glu concentration increased to 20mg / L, the concentration of L-isoleucine did not change significantly, considering the increase in production and economic factors , choose the most appropriate content of 20mg / L.

[0059] Set the time of fermentation culture as 24, 36, 48, 60, 72, 84 (h), pass figure 2 Visible, along with the increase of fermentation time, the concentration...

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Abstract

The invention discloses a production and extraction process of L-isoleucine, which comprises the following steps: step 1) fermentation, step 2) filter sterilization, step 3) chromatographic separation, step 4) concentration, and step 5) crystallization , step 6) drying, step 7) waste disposal. The present invention adopts steps such as ceramic membrane-decolorizing membrane, adsorption resin chromatography, double-cone rotary vacuum drying, etc. to obtain L-isoleucine high-quality goods, with high degree of automation, simple operation and labor saving.

Description

technical field [0001] The invention belongs to the field of biotechnology, relates to the technical field of L-isoleucine production, in particular to a production and extraction process of L-isoleucine. Background technique [0002] L-Isoleucine Molecular Formula C 6 h 13 NO 2 , the relative molecular mass is 131.17, and the melting point is 285°C-286°C. The specific rotation of L-isoleucine is [α]20D=+41.1°(6NHCl, C=4), and its dissociation constants pK1(α-COOH) and pK2(α-NH 3 + ) were 2.36 and 9.68, respectively; the isoelectric point (pI) was 6.02. L-isoleucine is soluble in water, hardly soluble in ethanol and ether, almost insoluble in other organic solvents, and the solubility in water increases with the increase of temperature. [0003] L-isoleucine is an important part of living organisms and one of the essential amino acids for the human body. It has a variety of physiological functions and plays an important role in many aspects such as metabolic regulation...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/06C07C227/40C07C229/08C12R1/15
CPCC07C227/40C12P13/06C07C229/08
Inventor 张宗华刘闪文包鑫李江雷韦树高崔国梁
Owner XINJIANG FUFENG BIOTECH
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