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Arthrobacter for producing beta-fructofuranosidase and application of arthrobacter

A technology of fructofuranosidase and Arthrobacter, which is applied in the direction of glycosylase, enzyme, bacteria, etc., can solve the problem of low yield of lactulose oligosaccharides, achieve the effect of reducing production cost and improving production capacity

Active Publication Date: 2016-11-23
SHANDONG BAILONG CHUANGYUAN BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Enzymatic synthesis is the main way to industrially produce lactulose-oligosaccharides. The low yield of enzymatic synthesis of lactulose-oligosaccharides has always been an important problem restricting the development of enzymatic synthesis. Therefore, it is necessary to find a β-fructofuranoside with high enzymatic activity. Enzyme becomes the key to solve the bottleneck of lactulose oligosaccharide production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A strain of Arthrobacter sp. BLCY-004 was deposited on August 16, 2016 in the General Microbiology Center of China Committee for the Collection of Microbial Cultures, address: Microbiological Research, Chinese Academy of Sciences, No. 1, Yard 1, Beichen West Road, Chaoyang District, Beijing Institute, deposit number CGMCC No.12855.

[0033] The screening process of the above-mentioned Arthrobacter sp. BLCY-004 is as follows:

[0034] (1) Enrichment culture

[0035] Select the soil near the production workshop of Bailong Chuangyuan in Dezhou, Shandong, remove the topsoil with a small shovel, take about 10g of soil 5-15cm above the ground, dilute it 10 times with sterile water, add the medium for enrichment culture, and the composition of the medium : Glucose 2%, yeast extract 1.5%, potassium dihydrogen phosphate 0.2%, diammonium hydrogen phosphate 0.6%, magnesium sulfate heptahydrate 0.01%, pH 6.5; culture temperature 28-35 ℃, culture 36h.

[0036] (2) purebred separat...

Embodiment 2

[0045] The culture method of the Arthrobacter sp. BLCY-004 described in embodiment 1, the steps are as follows:

[0046] (1) Arthrobacter sp. BLCY-004 was inoculated in LB medium, and activated and cultured for 30 hours at 30°C to obtain an activated strain;

[0047] (2) Take the activated bacterial strain prepared in step (1), inoculate it in the seed culture medium, and proliferate and cultivate it for 36 hours at 32° C. to obtain the seed solution;

[0048] Described seed culture medium component is as follows, is weight percent:

[0049] Glucose 2%, yeast extract 1.5%, potassium dihydrogen phosphate 0.2%, diammonium hydrogen phosphate 0.6%, magnesium sulfate heptahydrate 0.01%, pH6.5.

[0050] (3) Take the seed solution prepared in step (2), inoculate it in the fermentation medium at a ratio of 1% by volume, and expand the cultivation for 35 hours at 30° C. to obtain the bacterial cell fermentation solution;

[0051] Described fermentation medium component is as follows,...

Embodiment 3

[0055] The culture method of the Arthrobacter sp. BLCY-004 described in embodiment 1, the steps are as follows:

[0056] (1) Take Arthrobacter sp. BLCY-004 and inoculate it in LB medium, and activate it for 36 hours at 31°C to obtain an activated strain;

[0057] (2) Take the activated bacterial strain prepared in step (1), inoculate it in the seed culture medium, and proliferate and cultivate it for 36 hours at 32° C. to obtain the seed solution;

[0058] Described seed culture medium component is as follows, is weight percent:

[0059] Glucose 2%, yeast extract 1.5%, potassium dihydrogen phosphate 0.2%, diammonium hydrogen phosphate 0.6%, magnesium sulfate heptahydrate 0.01%, pH 7.0.

[0060] (3) Take the seed liquid prepared in step (2), inoculate it in the fermentation medium at a ratio of 5% by volume, and expand the cultivation for 40 hours at 30° C. to obtain the bacterial cell fermentation liquid;

[0061] Described fermentation medium component is as follows, is wei...

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Abstract

The invention relates to arthrobacter for producing beta-fructofuranosidase and application of the arthrobacter. The Arthrobacter sp. BLCY-004 is preserved in the China General Microbiological Culture Collection Center on August 16th, 2016, the address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, No.1 Yard, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No. 12855. The arthrobacter is separated from soil, the high-producing strain of high-producing beta-fructofuranosidase is obtained by means of mutagenic treatment technologies such as ultraviolet mutagenesis, nitrosoguanidine mutagenesis and the like and is named as BLCY-004, the enzyme activity reaches 1100U / ml and is improved by more than 50% as compared with that of conventional beta-fructofuranosidase, the capacity that sucrose is converted into lactosucrose can be highly improved by applying the arthrobacter to production of lactosucrose, and production cost is lowered significantly.

Description

technical field [0001] The invention relates to the production of a β-fructofuranosidase-producing Arthrobacter and its application, and belongs to the technical field of biotechnology. Background technique [0002] Lactulooligosaccharide is a functional oligosaccharide, which has special physiological functions such as promoting the proliferation of intestinal bifidobacteria, regulating intestinal microecology, and improving intestinal immunity. The sweetness is 30% of sucrose, and the sweetness is similar to that of sucrose Compared with sucrose, the sweetness quality is the best among various oligosaccharides, so it can be used in the food industry without worrying about its impact on product flavor. Commercially produced lactulose-oligosaccharides are slightly sweeter because they contain other ingredients such as sucrose and lactose. Compared with other oligosaccharides, oligosaccharides have higher stability to acid and heat. Its stability is similar to that of sucros...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/26C12R1/06
CPCC12N1/20C12N9/2431C12Y302/01026C12N1/205C12R2001/06
Inventor 干昭波窦光朋杜倩王艳辉邵先豹李方华
Owner SHANDONG BAILONG CHUANGYUAN BIO TECH
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