Method for producing L-tryptophan

A production method, tryptophan technology, applied in the field of microbial fermentation, can solve the problems of breeding technology, backward production technology, less research on L-tryptophan fermentation, low conversion rate and extraction comprehensive yield, and achieve production cost Low content, small content fluctuation, and stable product quality

Active Publication Date: 2010-12-15
SHANDONG YANGCHENG BIOLOGY TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the 1930s, Japan carried out large-scale research on the production of L-tryptophan by fermentation, mainly focusing on traditional mutation breeding techniques to change the characteristics of strains and increase the production of L-tryptophan, up to 7.2g / L. Few researches on L-tryptophan fermentation in recent years
The main reason is that the breeding technology and production technology are backward, the acid production level of the strain is not high; the fermentation cycle is long, the conversion rate and the comprehensive yield of extraction are low

Method used

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  • Method for producing L-tryptophan

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] 1. Shake flask liquid culture:

[0047] ①Put the primary strain culture medium into eggplant bottles, then inoculate the Corynebacterium glutamicum strain at an inoculum amount of 0.1%, and obtain the primary strain after static cultivation at 32°C for 24 hours;

[0048] ②Put the shake flask culture medium into the shake flask, insert the primary strain culture medium solution obtained in the previous step, and cultivate it on a reciprocating shaker with an inoculum size of 0.1%, a rotation speed of 120 rpm, a temperature of 32°C, and a time period of 18 hours. , to obtain shake flask liquid strains, the obtained strains were uniform, strong and non-polluting under the microscope, measured with a 752 spectrophotometer (range: 10mm, wavelength: 600nm) and the OD was 0.68.

[0049] 2. Production of glucose solution:

[0050] ① Soak the corn in the soaking tank for 4 hours and maintain the temperature at 50°C;

[0051] ② Grind the corn into corn milk slurry with a sand m...

Embodiment 2

[0066] 1. Shake flask liquid culture:

[0067] ①Put the primary strain culture medium into the eggplant bottle, then inoculate the Corynebacterium glutamicum strain according to the inoculum amount of 0.2%, and obtain the primary strain after static cultivation at 30-33°C for 20-24 hours;

[0068] ②Put the shake flask culture medium into the shake flask, insert the primary strain culture medium obtained in the previous step, and cultivate it on a reciprocating shaker with an inoculum size of 0.15%, a rotation speed of 140 rpm, a temperature of 30-33°C, and a time of After a period of 16 hours, the liquid strains in shake flasks were obtained. Microscopically, the obtained strains were uniform, strong, and pollution-free. Measured with a 752 spectrophotometer (range: 10mm, wavelength: 600nm), the OD was 0.6.

[0069] 2. Production of glucose solution:

[0070] ① Soak the corn in the soaking tank for 3-5 hours, and the soaking temperature is 55°C;

[0071] ②Use a sand mill to ...

Embodiment 3

[0086] 1. Shake flask liquid culture:

[0087] ①Put the primary strain culture medium into eggplant bottles, and then inoculate the Corynebacterium glutamicum strain according to the inoculum amount of 0.15%, and obtain the primary strain after static cultivation at 30-33°C for 20-24 hours;

[0088] ②Put the shake flask culture medium into the shake flask, insert the primary strain culture medium solution obtained in the previous step, and cultivate it on a reciprocating shaker with an inoculum size of 0.2%, a rotation speed of 130 rpm, a temperature of 30-33°C, and a time of After a period of 16 hours, the liquid strains in shake flasks were obtained. Microscopically, the obtained strains were uniform, strong, and pollution-free. Measured with a 752 spectrophotometer (range: 10mm, wavelength: 600nm), the OD was 0.8.

[0089] 2. Production of glucose solution:

[0090] ① Soak the corn in the soaking tank for 3-5 hours, and the soaking temperature is 53°C;

[0091] ②Use a san...

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Abstract

The invention discloses a method for producing L-tryptophan, which comprises the following steps of: performing primary culture and shaking culture on corynebacterium glutamicum strains to obtain shaking liquid strains; preparing glucose liquid from corn serving as a raw material by steps of soaking, pulping, liquefying, saccharifying and the like, wherein the glucose liquid is used as a carbon source of a culture medium and adds sugar for fermentation flow; performing secondary culture on the shaking liquid strains to obtain secondary strains, and performing fermentation culture on the secondary strains to obtain L-tryptophan fermentation liquid; and extracting and refining the L-tryptophan fermentation liquid to obtain the L-tryptophan. The acid yield of the strains can reach 40g / L at most by controlling the composition of the culture medium and the fermentation conditions; the glucose liquid is produced by using the corn, so the production cost is low; the fluctuation of the glucose content of the culture medium is low and the saccharic acid conversion rate of a fermentation acid production level box is improved by continuously adding the glucose liquid; and the extracting and refining process flows are reasonable, the purity of the L-tryptophan product can reach over 99 percent, and the yield reaches over 75 percent.

Description

technical field [0001] The invention relates to an amino acid production method, in particular to a fermentation production method of L-tryptophan, which belongs to the technical field of microbial fermentation. Background technique [0002] L-Tryptophan is a neutral aromatic amino acid containing indolyl, scientific name: β-indolylalanine; English name: Tryptophan; molecular formula: C 11 h 12 N 2 o 2 , the relative molecular mass is 204.21, and the melting point is 289°C. Its structural formula is as follows: [0003] [0004] L-Tryptophan is white or light yellow crystal or crystalline powder; no odor and slightly bitter. Slightly soluble in water, very slightly soluble in ethanol, insoluble in chloroform, easily soluble in formic acid, soluble in sodium hydroxide solution or dilute hydrochloric acid. It widely exists in natural proteins. In addition to playing an important role in nutritional metabolism, L-tryptophan is also widely used in medicine, food, feed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/22C07D209/20C12R1/15
Inventor 王东阳蔡传康闫汝东冯志彬张华
Owner SHANDONG YANGCHENG BIOLOGY TECH CO LTD
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