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Method for producing chitosan oligosaccharide through solid-state fermentation of aspergillus oryzae

A solid-state fermentation, Aspergillus oryzae technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problem of low yield of chitosan

Pending Publication Date: 2019-12-27
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield of chitosan produced by chitosan enzymatic method is low, and the fermentation conditions need to be optimized

Method used

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  • Method for producing chitosan oligosaccharide through solid-state fermentation of aspergillus oryzae
  • Method for producing chitosan oligosaccharide through solid-state fermentation of aspergillus oryzae
  • Method for producing chitosan oligosaccharide through solid-state fermentation of aspergillus oryzae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Optimization of moisture content in the culture medium

[0050] On the basis of the fermented solid medium, add distilled water to the medium according to the initial water content of 30%, 40%, 50%, 60%, and 70% respectively (the water content of bran is calculated as 12%), and the incubator at 30°C Cultured in medium for 120h, samples were taken every 24h. The content of glucosamine, the relative enzyme activity of chitosanase and the number of spores were determined respectively. The result is as Figure 1-2 Shown, followed by the content of chitosan oligosaccharides, chitosanase relative enzyme activity changes in the fermentation process.

[0051] Depend on Figure 1-2 It can be seen that the output of chitosan oligosaccharide fluctuates with the increase of time. Because when generating chitosan oligosaccharide, the Aspergillus oryzae thallus in the culture medium also consumes a part of chitosan oligosaccharide, so the output of chitosan oligosaccha...

Embodiment 2

[0054] Embodiment 2: inoculum size optimization

[0055] After confirming that the above-mentioned initial water content is 50%, on the basis of fermentation solid medium, inoculate according to the inoculation amount of 5%, 10%, and 15%, respectively, cultivate in a 30°C incubator for 120h, and take a sample every 24h. The content of glucosamine and the activity of chitosanase were determined respectively. The result is as Figure 3-4 Shown, followed by the content of chitosan oligosaccharides, chitosanase relative enzyme activity changes in the fermentation process.

[0056] Depend on Figure 3-4 It can be seen that the oligochitosan production in the 5% culture medium increases gradually with time, and the maximum value is 2.55 mg / ml for 120 h of fermentation; the 10% culture medium increases first Afterwards, the decline is increasing, and its maximum value is 2.36 mg / ml when it is fermented for 96 hours; the production of chitosan oligosaccharides in the 15% culture me...

Embodiment 3

[0059] Embodiment 3: Substrate addition amount optimization

[0060] After determining that the above-mentioned initial moisture content is 50%, and the inoculum size is 15%, on the basis of the fermentation solid medium, add chitosan according to the substrate addition amount of 2%, 5%, 8%, and 10%, respectively, at 30°C Cultured in the incubator for 120h, and samples were taken every 24h. The content of glucosamine and the activity of chitosanase were determined respectively. The result is as Figure 5-6 Shown, followed by the content of chitosan oligosaccharides, chitosanase relative enzyme activity changes in the fermentation process.

[0061] Depend on Figure 5-6 It can be seen that the production of chitosan oligosaccharides fluctuates with time in the medium with different additions of four kinds of substrates, and they all have a trend of first increasing, then decreasing, then increasing and then decreasing. The four different gradient media all reached the highe...

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Abstract

The invention discloses a method for producing chitosan oligosaccharide through solid-state fermentation of aspergillus oryzae. According to the method, firstly, aspergillus oryzae strains stored intoa test tube are activated and are transferred into a conical flask, and are then inoculated into a solid-state culture medium; the aspergillus oryzae is subjected to solid-state fermentation cultureunder the culture condition of the fermentation temperature being 30 DEG C; after the fermentation for 4d, the fermentation is terminated; and a fermentation product containing the chitosan oligosaccharide is obtained. The six conditions including the initial water content of the solid-state culture medium, the inoculation quantity, the substrate addition quantity, the fermentation temperature, the fermentation time and the substrate treatment are optimized; the chitosan oligosaccharide yield and the chitosanase activity are used as indexes to judging the optimum conditions; and finally, the result that the yield of the chitosan oligosaccharide obtained under the conditions of the initial water content of the culture medium being 50 percent, the inoculation quantity being 15 percent, the substrate addition quantity being 2 percent, the fermentation temperature being 30 DEG C and the fermentation time being 4d is highest is obtained. The method provided by the invention is suitable forindustrialized production of the high-content chitosan oligosaccharide.

Description

technical field [0001] The invention belongs to the technical field of aspergillus oryzae fermentation, and in particular relates to a method for producing chitosan oligosaccharide by solid-state fermentation of aspergillus oryzae. Background technique [0002] Aspergillus oryzae (Aspergillus oryzae) belongs to the phylum Ascomycota, the order Aspergillus, the family Aspergillus, the genus Aspergillus, and the Aspergillus flavus group. It is a type of mold widely used in winemaking and fermentation industries. It has multiple enzyme systems such as protease enzyme system with strong protein decomposition ability and saccharification enzyme system with strong starch saccharification ability. Aspergillus oryzae is easy to cultivate and does not produce aflatoxin, so it is a kind of mold with high utilization value. Aspergillus oryzae is one of the safe strains recognized by the US Food and Drug Administration. Aspergillus oryzae has been widely used in food industry, agricul...

Claims

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Application Information

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IPC IPC(8): C12P19/26C12P19/14C12R1/69
CPCC12P19/26C12P19/14
Inventor 孙建安毛相朝赵甜宇
Owner OCEAN UNIV OF CHINA
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