Method for catalytic production of fructo-oligosaccharides by aspergillus oryzae whole cells

A technology of cane fructose oligosaccharide and whole cell catalyst, applied in the field of oligosaccharide, can solve the problems of harsh production environment and conditions, complicated separation and purification process, and high cost of enzymatic reaction, and achieves easy operation, simple process and simplification. The effect of the production process

Active Publication Date: 2013-07-10
广东比克生物科技有限公司
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Problems solved by technology

The disadvantage of this method is that there are many steps, the enzyme is easily inactivated, the conversion rate is low, and the uniformity of mass transfer is limited because sucrose and converted products have to pass through the immobilized particles.
[0004] Due to the complexity of the separation and purification process of the enzyme and the partial loss of the enzyme activity during the process, the instability in the three-dimensional structure environment, and the disadvantages of easy inactivation, and the high price of the enzyme, resulting in high cost of the enzymatic reaction, these aspects are serious. restricts its industrial application
[0005] Immobilized cell method, that is, the method of embedding enzyme-producing bacteria with calcium alginate gel. The disadvantage of this method is that the production environment and conditions are harsh (such as air purification)

Method used

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  • Method for catalytic production of fructo-oligosaccharides by aspergillus oryzae whole cells

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Embodiment 1

[0029] The first step is to inoculate the spores of Aspergillus oryzae CICC 2134 (Aspergilusoryzae2134, preserved by the China Industrial Microorganism Culture Collection Management Center, address: No. 32 Xiaoyun Road, Chaoyang District, Beijing, Zip Code: 100027) into the liquid seed medium. 1×10 5 A / L, at 33 ℃, the thalline solution after 16h of cultivating on the shaker of rotating speed 150r / min is made seed liquid; The formula of described liquid seed culture medium is: 20g / L sucrose, 30g / L corn flour, 100g / L L yeast extract powder, 2g / L NaNO 3 ;Stir and dissolve evenly, then sterilize, and cool for later use;

[0030] Step 2: Inoculate the seed solution obtained in the first step into the liquid medium for producing enzyme inducer at a ratio of 5:100 by volume, and cultivate the bacteria for 24 hours at 33°C on a shaker with a rotation speed of 150rpm The solution is made into a whole-cell thalline solution; the liquid medium containing an enzyme inducer is: 70g / L suc...

Embodiment 2

[0036] The first step is to inoculate the spores of Aspergilus oryzae CICC 2134 (Aspergilus oryzae 2134, preserved by China Industrial Microorganism Culture Collection Management Center, address: No. 32 Xiaoyun Road, Chaoyang District, Beijing, Zip Code: 100027) into the liquid seed medium, The inoculum size is 1×10 10 A / L, at 28 ℃, the bacterial cell solution after 24 hours of cultivating on a shaker with a rotating speed of 100r / min is made into a seed liquid; the formula of the liquid seed medium is: 50g / L sucrose, 7g / L malt powder, 10g / L L peptone, 3g / L NaCl; stir and dissolve evenly, use for sterilization, cool and set aside;

[0037] The second step: Inoculate the seed solution obtained in the first step into the liquid medium for producing enzyme inducer at a volume ratio of 10:100, and cultivate the bacteria for 16 hours at 28°C with a rotation speed of 100rpm The solution is made into a whole-cell thalline solution; the liquid culture medium containing an enzyme indu...

Embodiment 3

[0042] The first step is to inoculate the spores of Aspergilus oryzae CICC 2134 (Aspergilus oryzae 2134, preserved by China Industrial Microorganism Culture Collection Management Center, address: No. 32 Xiaoyun Road, Chaoyang District, Beijing, Zip Code: 100027) into the liquid seed medium, The inoculum size is 1×10 6 A / L, at 25 ℃, the bacterial cell solution after 20h of cultivating on the shaker of 120r / min rotating speed is made seed liquid; The formula of described liquid seed culture medium is: 30g / L sucrose, 5g / L malt powder, 7g / L L peptone, 50g / L yeast extract, 1.5g / L NaCl, 1.5g / L NaNO 3 ;Stir and dissolve evenly, then sterilize, and cool for later use;

[0043] The second step is to inoculate the seed solution obtained in the first step into the liquid medium for producing enzyme inducer with a volume ratio of 8:100, and cultivate the bacterial cell solution for 20 hours at 25°C on a shaker with a rotation speed of 300rpm Make whole cell thalline solution; Described ...

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Abstract

The invention discloses a method for catalytic production of fructo-oligosaccharides by aspergillus oryzae whole cells. The method comprises the following steps of 1, whole-cell catalyst preparation comprising inoculating a seed medium with aspergillus oryzae spores, preparing a seed solution under a certain condition, inoculating a liquid fermentation enzyme-production medium for producing an enzyme inducer with the seed solution, preparing a whole-cell aspergillus oryzae solution under a certain condition, collecting aspergillus oryzae, filtering the aspergillus oryzae by a filter cloth of 200 to 400 meshes, and carrying out washing, re-filtration and vacuum freeze drying to obtain aspergillus oryzae whole cells, and 2, aspergillus oryzae whole cell-based fructo-oligosaccharide catalytic production comprising carrying out conversion under a certain conditions and carrying out filtration and vacuum concentration of the obtained fructo-oligosaccharide liquid to obtain fructo-oligosaccharide syrup. A high pressure liquid chromatography test on the fructo-oligosaccharide product proves that the solids comprise greater than or equal to 50% of total oligosaccharides. The method has simple processes, is convenient for operation, and has high enzyme activity, high conversion efficiency and an important industrial value.

Description

technical field [0001] The invention belongs to the technical field of oligosaccharides, and relates to a fermentation production method of fructooligosaccharides, in particular to a method for producing fructooligosaccharides from whole cells of Aspergillus oryzae. Background technique [0002] Oligosaccharides are carbohydrates with a low degree of polymerization formed by connecting 2-10 monosaccharide molecules through glycosidic bonds. Among them, functional fructooligosaccharides, also known as fructooligosaccharides (FOS for short), are From 1 to 3 fructosyl groups combined with sucrose through β-1,2 glycosides, a mixture of kestose, kestose and kestose pentaose is generated. FOS has excellent properties such as promoting the proliferation of bifidobacteria in the intestinal tract, detoxifying and cleaning the intestinal tract, improving human immunity, improving lipid metabolism, preventing tooth decay, and not being absorbed and utilized by the digestive tract. It i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/18C12R1/69
Inventor 刘冬梅周康范梦柯叶嘉伦韦健军周伟文韦胜军
Owner 广东比克生物科技有限公司
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