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A β-fructofuranosidase-producing Arthrobacter and its application

An Arthrobacter, microbial strain technology, applied in the direction of glycosylase, enzyme, bacteria, etc., can solve the problems of low yield of lactulose oligosaccharide, etc., and achieve the effect of reducing production cost and improving ability

Active Publication Date: 2019-03-19
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 a β-fructofuranosidase-producing Arthrobacter and its application. A strain of Arthrobacter sp. BLCY‑004 was deposited on August 16, 2016 at 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. The present invention isolates Arthrobacterium from soil, and after undergoing mutagenesis treatment techniques such as ultraviolet mutagenesis and nitrosoguanidine mutagenesis treatment, the high-yield strain with high production of β-fructofuranosidase is finally obtained and named BLCY-004, and its enzyme activity It reaches 1100U / ml, which is more than 50% higher than the activity of traditional β-fructofuranosidase. It can greatly improve the ability of sucrose to be converted into lactulose-oligosaccharides when used in the production of lactulose-oligosaccharides, and significantly reduce production costs.

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 Patents(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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