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Production and extraction technology of L-isoleucine

A technology of isoleucine and extraction process, applied in the direction of biochemical equipment and methods, microorganisms, chemical instruments and methods, etc., can solve the problems of reagent consumption, production waste, increase process complexity and workload, etc., and achieve product saving Cost, increase fermentation yield, small footprint

Active Publication Date: 2019-04-16
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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  • Production and extraction technology of L-isoleucine
  • Production and extraction technology of L-isoleucine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A production and extraction process of L-isoleucine, which comprises the following steps:

[0036] Step 1): Cultivate Corynebacterium glutamicum (CGMCC NO.12153, donated by Tianjin University of Science and Technology) to a concentration of 5×10 8 The CFU / mL seed solution was then inoculated into the fermentation medium according to 10% inoculation amount, and fermented for 72 hours. The fermentation culture conditions were: temperature 30°C, dissolved oxygen 30%, pH 6.5-6.8, to obtain L- Isoleucine fermentation broth; the components of the fermentation medium are: glucose 120 g / L, (NH 4 ) 2 SO 4 20g / L, FeSO 4 ·7H 2 O 0.015 g / L, MgSO 4 ·7H 2 O 0.4 g / L, MnSO 4 ·H 2 O 0.015 g / 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;

[0037]Step 2): Pass the L-isoleucine fermentation broth through a ceramic membrane (20nm) to remove bacterial protein; membrane filtration operating temperature: 45°C, workin...

Embodiment 2

[0045] A production and extraction process of L-isoleucine, which comprises the following steps:

[0046] Step 1): Cultivate Corynebacterium glutamicum (CGMCC NO.12153, donated by Tianjin University of Science and Technology) to a concentration of 5×10 8 CFU / mL seed liquid, then inoculated into the fermentation medium according to 10% inoculation amount, and fermented for 75 hours. The fermentation culture conditions were: temperature 30°C, dissolved oxygen 30%, pH 6.5-6.8, to obtain L- Isoleucine fermentation broth; the components of the fermentation medium are: glucose 160 g / L, (NH 4 ) 2 SO 4 30 g / L, FeSO 4 ·7H 2 O 0.015 g / L, MgSO 4 ·7H 2 O 0.4 g / L, MnSO 4 ·H 2 O 0.015 g / 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;

[0047] Step 2): Pass the L-isoleucine fermentation broth through a ceramic membrane (50nm) to remove bacterial protein; membrane filtration operating temperature: 50°C, working press...

Embodiment 3

[0055] Taking 200L fermentation medium as an example, a total of 6.53kg of L-isoleucine product was obtained. The appearance of the product is white and translucent. The purity is 99.4%.

[0056] The present invention also detects the influence of culture medium and culture time on the concentration of L-isoleucine in the fermentation broth.

[0057] Taking Example 1 as the experimental group, set the Glu concentration in the medium to 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.

[0058] 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 increase of L...

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Abstract

The invention discloses a production and extraction technology of L-isoleucine. The technology comprises the steps of 1), fermentation; 2), filtering and bacteria removing; 3), chromatographic fractionation; 4), concentration; 5), crystallization; 6), drying; 7), waste treatment. According to the production and extraction technology, L-isoleucine essence is obtained through the steps of ceramic membrane-decoloring membrane, resin chromatography absorbing, double conical rotary vacuum drying and the like, the automation level is high, the operation is simple, and labor cost is saved.

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; 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 and informati...

Claims

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Application Information

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Patent Type & Authority Applications(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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