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Strain and method for producing glucosamine through microorganism fermentation

A glucosamine and microbial fermentation technology applied in the biological field to achieve low cost, short production cycle, and stable production and supply

Active Publication Date: 2015-11-11
安徽福方高科生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to provide a method for producing N-acetyl-D-glucosamine and D-glucosamine by microbial fermentation, so as to overcome the above-mentioned deficiencies in the prior art

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  • Strain and method for producing glucosamine through microorganism fermentation
  • Strain and method for producing glucosamine through microorganism fermentation

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Embodiment 1

[0043] According to another aspect of the present invention, the D-glucosamine method of producing N-acetyl-D-glucosamine and D-glucosamine by fermentation of the above bacterial strains is used as the starting strain to produce N-acetyl-D-glucosamine through seed culture and optimized medium fermentation. Acetyl-D-glucosamine and D-glucosamine, including the following steps:

[0044] (1) Strain screening, identification and cultivation

[0045] Collect 50 soil samples, dilute them 10-3 times and spread them on the primary screening plate. Use the primary screening medium: colloidal chitin 2.5g / L, dipotassium hydrogen phosphate 0.7g / L, potassium dihydrogen phosphate 0.3g / L L, Magnesium Sulfate 0.5g / L, Ferrous Sulfate 0.01g / L, Agar 20g / L, pH 7.0, culture temperature 37°C, culture time 72h, cultured, obtained a single colony, separated the colony, and obtained 11 transparent colonies The strains with large and bright circles and good growth conditions were identified according ...

Embodiment 2

[0078] (1) UV radiation-induced mutation

[0079] Take 10ml1×10 7 individual / ml Bacillus subtilis ( Bacillus subtilis ) NJ090259 suspension was placed in a 9cm-diameter petri dish, and the ultraviolet lamp was preheated for 20 minutes. The petri dish was placed on a magnetic stirrer and the vertical distance of the petri dish was about 30cm from a 10W ultraviolet lamp; turn on the magnetic stirrer and irradiate 150, 200, 250, 300s; after the mutagenesis of the bacteria solution, let it stand in the refrigerator for 1-2 hours in the dark, take the mutagenized strains and the starting strains and inoculate them on the chitin medium plate respectively, and select the chitin medium plates with fast growth speed. The ratio of hydrolysis circle to colony diameter is greater than 10% of the starting bacteria and the largest mutant is tested for enzyme activity;

[0080] Chitin medium: colloidal chitin 30g / L, ammonium sulfate 2.0g / L, magnesium sulfate 0.5g / L, potassium dihydrogen p...

Embodiment 3

[0089] (1) Bacillus subtilis ( Bacillus subtilis ) NJ090259 UV-induced mutant fed-feed fermentation test

[0090] UV-induced Bacillus subtilis ( Bacillus subtilis ) After the largest mutant of NJ090259 was placed on the plate medium for activation, it was inserted into the seed medium and cultured in a constant temperature shaking table at 30°C for 18 hours, which was used as the seed liquid; when inoculated, it was inserted into 50ml of fermentation culture at a ratio of 1:10 In the base 250mL baffled Erlenmeyer flask, add 2.5ml of feed medium at 24h, 36h, 48h, and 60h respectively;

[0091] Fermentation medium: colloidal chitin 10g / L, glucose 10g / L, yeast extract 3.0g / L, MgSO 4 ·7H 2 O0.6g / L, FeSO 4 ·7H 2 O0.01g / L, KH 2 PO 4 0.4g / L, K 2 HPO 4 0.6g / L, ZnSO 4 0.001g / L;

[0092] Feed medium: colloidal chitin 100g / L, glucose 100g / L, pH 6.0.

[0093] Culture conditions: pH 6.5, culture temperature 35°C, constant temperature shaker culture, culture time 72h.

[0094...

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Abstract

The invention relates to a non-genetic recombinant strain for producing N-acetyl-D-glucosamine and D-glucosamine through microorganism fermentation, wherein the strain has been preserved in China General Microbiological Culture Collection Center. A bacillus subtilis strain NJ090259 is assigned the accession number of CGMCC10257. A bacillus licheniformis strain NJ091195 is assigned the accession number of CGMCC10258. The two strains are both preserved at 29th, December, 2014. The invention provides the novel strain for producing N-acetyl-D-glucosamine and D-glucosamine and the production method thereof. By means of the method, stable production and supplement of the N-acetyl-D-glucosamine can be achieved. The invention also achieves non-animal-sourced and safe production of the N-acetyl-D-glucosamine and the D-glucosamine. The method is short in production period, is low in cost and is more environment-friendly.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a bacterial strain for producing N-acetyl-D-glucosamine and D-glucosamine by microbial fermentation and a method thereof. Background technique [0002] N-acetyl-D-glucosamine is a monosaccharide, which is the chitin in the cell wall of fungi (basidiomycete, mold or yeast), or the composition of crustaceans such as crabs and shrimp shells, and can also be used as a food with a very high content. less nutrients. N-acetyl-D-glucosamine has a similar effect to glucosamine. Taking a certain amount of N-acetyl-D-glucosamine can induce the production of new cartilage and inhibit the onset of osteoarthritis. In some cases, N - Acetyl-D-glucosamine is also used to treat osteoarthritis. Since glucosamine has a bitter taste, while N-acetyl-D-glucosamine has 50% of the sweetness of sucrose and is easy to ingest, N-acetyl-D-glucosamine has received extensive attention as a substitute for gluc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/26C12R1/125C12R1/10
CPCC12P19/28C12R2001/10C12N1/205C12R2001/125C12P19/26
Inventor 邹季虹
Owner 安徽福方高科生物科技有限公司
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