A kind of method of industrialized fermentation high-yield l-tryptophan

A tryptophan, high-yield technology, applied in the field of amino acid fermentation, can solve the problems of low production cost in the fermentation cycle, low L-tryptophan acid production rate, etc., to achieve easy operation and maintenance, obvious decolorization effect, and less floor space Effect

Active Publication Date: 2017-12-29
XINJIANG FUFENG BIOTECH
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention solves the problem of low acid production rate of L-tryptophan in the industrialized production of biological fermentation. The purpose of the present invention is to provide a method for high-yield L-tryptophan in industrial fermentation. High and low production cost, stable process and strong practicability

Method used

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  • A kind of method of industrialized fermentation high-yield l-tryptophan

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Experimental program
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Effect test

Embodiment 1

[0025] A method for industrialized fermentation of high-yield L-tryptophan, comprising the steps of:

[0026] Step 1) Fermentation: Escherichia coli (Escherichiacoli) ATCC 27325 (see Appl.Environ.Microbiol.October 1994vol.60no.10, 3724-3731) with tnaA, trpR and tyrR gene knockout or inactivation and glutamic acid rod Corynebacterium glutamicum ATCC 13032 (Journal of Biotechnology, V104, September 2003, Pages5-25) was cultured to a concentration of 1×10 7 Each / mL of the bacterial liquid, mixed according to the volume ratio of 1:1 to obtain the mixed bacterial liquid, then inoculated into the fermentation medium (glucose 20g / L, yeast powder 15g / L, magnesium sulfate according to 5% (v / v) inoculum 2.5g / L, Ammonium Sulfate 2g / L, Citric Acid 2.5g / L, Dipotassium Hydrogen Phosphate 9g / L, Ferrous Sulfate 76mg / L, Manganese Sulfate 5mg / L, Sodium Sulfate 20mg / L, Zinc Sulfate 7mg / L , cobalt chloride 6mg / L, copper sulfate 0.9mg / L), temperature 37°C, dissolved oxygen control 20%, tank press...

Embodiment 2

[0034] A method for industrialized fermentation of high-yield L-tryptophan, comprising the steps of:

[0035] Step 1) Fermentation: Escherichia coli (Escherichia coli) CCTCC M 2011316 (see CN201110400855X) and Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC13032 were cultivated to a concentration of 1×10 7 Each / mL of the bacterial liquid, mixed according to the volume ratio of 1:1 to obtain the mixed bacterial liquid, then inoculated into the fermentation medium (glucose 20g / L, yeast powder 15g / L, magnesium sulfate according to 5% (v / v) inoculum 2.5g / L, Ammonium Sulfate 2g / L, Citric Acid 2.5g / L, Dipotassium Hydrogen Phosphate 9g / L, Ferrous Sulfate 76mg / L, Manganese Sulfate 5mg / L, Sodium Sulfate 20mg / L, Zinc Sulfate 7mg / L , cobalt chloride 6mg / L, copper sulfate 0.9mg / L), temperature 37°C, dissolved oxygen control 20%, tank pressure 0.05MPa, pH 7.0, when the glucose in the fermentation medium is exhausted, it will enter the production of sugar supplementation In th...

Embodiment 3

[0043] The bacterial protein powder prepared in Example 1 was used to replace the yeast powder in the standard YPD medium, and the rest of the ingredients were unchanged. Saccharomyces cerevisiae and Pichia pastoris were cultivated under the same conditions, and the culture effect of the product was evaluated by comparing the growth of the cells. See Table 1.

[0044] Table 1

[0045]

[0046] Conclusion: OD600 was used to measure the growth of cells characterized by turbidimetry, indicating that this product can replace commercially available yeast powder products and achieve similar culture effects.

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Abstract

The invention belongs to the field of amino acids, and discloses a method for industrialized fermentation with high yield of L-tryptophan, comprising the following steps: step 1) fermentation, step 2) microfiltration, step 3) separation and ultrafiltration, step 4) concentration, Step 5) crystallization and step 6) preparation of cell protein powder. The method of the invention has the characteristics of short fermentation period, high acid production rate, low production cost, stable process and strong practicability.

Description

technical field [0001] The invention relates to a method for industrialized fermentation with high yield of L-tryptophan, which belongs to the field of amino acid fermentation. Background technique [0002] L-tryptophan, also known as L-tryptophan amino acid, L-β-(3-indolyl)-α-alanine, is one of the eight essential amino acids in human and animal life activities. Growth and metabolism play an important role, known as the second essential amino acid, widely used in medicine, food and feed industries. At present, the annual demand for tryptophan in the world market is more than 10,000 tons, and it is growing at a rate of 10% per year. The market prospect is very broad. [0003] The production methods of L-tryptophan include proteolysis, chemical synthesis, enzymatic conversion and microbial fermentation. Due to many problems in industrial production of proteolysis, chemical synthesis and enzymatic conversion, the application is limited. Recently, the production of L-tryptop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P39/00C12P13/22C12R1/19C12R1/15
Inventor 郭英熙李学朋刘家宁左善董爽
Owner XINJIANG FUFENG BIOTECH
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