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Screening of high-yield glycine strains and application thereof in nitrile compound conversion

A glycine and compound technology, applied in fungi, fermentation and other directions, can solve the problem of no mold and other problems, and achieve the effects of environmental friendliness, mild reaction conditions, convenient and fast product separation and application

Active Publication Date: 2011-08-24
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many strains used to produce glycine have been reported abroad, there are no molds, especially Fusarium oxysporum, that produce glycine.

Method used

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  • Screening of high-yield glycine strains and application thereof in nitrile compound conversion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Isolation and Identification of Fusarium oxysporum H3 and Preservation of the Strain

[0036] (1) Enrichment screening process of Fusarium oxysporum H3.

[0037] Collect soil samples around the nitrile compound production plant. When sampling, first remove the top soil with a small shovel, and collect soil samples at 5-15cm. Take 1g of soil sample, put it into an Erlenmeyer flask (20mL / 250mL) containing physiological saline, shake it vigorously for 10min, and let it stand still. Take 1mL of soil suspension, inject it on a sterile plate, and pour 25mL of sterilized and cooled to 40-50°C solid screening medium into the plate. After the culture medium was completely cooled and solidified, the plate was placed in a constant temperature incubator and cultured upside down at 30°C for 3 days. The composition of solid screening medium is: glucose 5.0g / L, potassium dihydrogen phosphate 1.0g / L, magnesium sulfate 0.1g / L, ferrous sulfate 0.02g / L, calcium chloride 0.02g / ...

Embodiment 2

[0055] Embodiment 2 Fusarium oxysporum (Fusarium oxysporum H3) fermentation method for producing glycine

[0056] This example illustrates the method of culturing Fusarium oxysporum H3 into a nutrient-rich medium to obtain bacteria with high activity and high biomass, and the method of biotransforming glycylnitrile to produce glycine.

[0057] The composition of the medium: glycerol 10.0g / L, yeast extract 1.0g / L, peptone 1.0g / L, magnesium sulfate 0.1g / L, potassium dihydrogen phosphate 1.0g / L, sodium chloride 0.5g / L, sulfite Iron 0.01g / L, caprolactam 0.5g / L, pH 7.5;

[0058] Insert Fusarium oxysporum H3 into the culture medium with 1% inoculum amount, and cultivate it for 3 days at 30°C and 120rpm. After the cultivation is over, collect the cultured cells by centrifuging at 12000rpm for 10min, and wash them with 0.5M phosphoric acid. The bacteria were washed twice with the buffer, and the bacteria were resuspended in the buffer to prepare the bacteria suspension.

[0059] Pre...

Embodiment 3

[0060] Example 3 Change of fermentation conditions, Fusarium oxysporum (Fusarium oxysporum H3) fermentation to produce glycine

[0061] This embodiment adopts the same method as in embodiment 2, changing the reaction time.

[0062] The bacteria were collected from the culture medium by centrifugation, washed twice with 0.5M phosphate buffer, resuspended to make a bacterial suspension, mixed with 7.8g / L glycyl nitrile solution, at 30°C, 120rpm After 12 hours of reaction, the concentration of glycine in the reaction liquid was 8.5 g / L, and the yield reached 80%.

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Abstract

The invention belongs to the biological technical field, and in particular relates to screening of high-yield glycine moulds and an application thereof in nitrile compound conversion. Glycine is widely applied to the fields such as food, chemical industry, medicine and the like; and the glycine is produced by utilizing a bioconversion method instead of a synthesis method, which becomes a development trend at home and abroad. The invention provides a screening method of high-yield glycine strains. By utilizing the method, a high-yield mould strain H3 for the glycine is screened for the first time, and the mould strain is identified as Fusarium oxysporum by combining morphological, physiological and biochemical characters and sequencing of genes, namely 18S, ITS and 28S rDNA; and the strain is conserved at China General Microbiological Culture Collection Center (conservation number: CGMCC 3829). The Fusarium oxysporum H3 screened by the method has the characteristics of fast growth speed, high conversion efficiency, short production cycle, mild reaction conditions, high yield, low production cost, environmental friendliness and the like, thus being applicable to industrial production of the glycine; and the after-treatment process of the product is convenient and fast without a large amount of acid or alkali.

Description

technical field [0001] A screening method for a high-yield glycine strain and its application. The invention relates to a method for producing glycine by using microorganisms to transform glycyl nitrile, which belongs to the field of biotechnology. Background technique [0002] Glycine, also known as aminoacetic acid, is a white odorless crystal, non-toxic, soluble in water, insoluble in organic solvents. It is the simplest structure among the 20 common amino acids, and it is a neutral aliphatic amino acid, which is a non-essential amino acid for the human body. Glycine is also used as the main raw material for the synthesis of glyphosate, the most widely used pesticide in the world, and its demand is very large; in addition, it can also be used as an intermediate in the preparation of food flavoring agents, flavor enhancers, high blood pressure, and liver damage. It is widely used in the fields of food, medicine and chemical industry. [0003] At present, the production o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P13/04C12P7/40
Inventor 许正宏张晓梅窦文芳许泓瑜龚劲松陆震鸣史劲松陈敬华
Owner JIANGNAN UNIV
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