Preparation method of transglucosidase

A technology of transglucosidase and spore suspension, which is applied in the field of preparation of transglucosidase, can solve the problems of low isomalto-oligosaccharide transglycosidase and the like, and achieve the effects of shortening reaction time, improving reaction rate and reducing production time.

Inactive Publication Date: 2016-04-20
SHANGHAI KINRY FOOD INGREDIENTS +1
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Problems solved by technology

[0007] The purpose of the present invention is to provide a method for preparing transglucosidase with a high rate of glucosidase conversion. Using this method, a crude solution o

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  • Preparation method of transglucosidase
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Example Embodiment

[0023] Example 1

[0024] Such as figure 1 As shown, a preparation method of transglucosidase, according to the production steps described in the content of the invention, aspergillus niger is cultivated on a slant for 7 days to prepare mature spores, and sterile water is added to prepare a slant spore suspension. In the PDY medium shake flask, culture at 32℃, 150r / min for 48h, and put the spore suspension in the shake flask in a 500ML shake flask containing 100ML liquid medium at a ratio of 10%. The weight percentages of the components of the optimized liquid fermentation medium are as follows: sucrose 10%, peptone 2%, yeast extract 0.3%, K 2 HPO 4 0.5%, N-methyl-D-glucosamine 0.05%, NaCl0.15%, MgSO 4 0.02%, the rest is water, prepared by mixing uniformly, initial pH 6.05, cultured at 28℃, 29℃, 30℃, 31℃, 32℃, and 180r / min for 60h, and centrifuged 5000r / min for fermentation broth , 2 minutes, take the supernatant to test the enzyme activity, as shown in Table 1. Enzyme activity ...

Example Embodiment

[0028] Example 2

[0029] Such as figure 1 As shown, a preparation method of transglucosidase, according to the production steps described in the content of the invention, aspergillus niger is cultivated on a slope for 7 days to prepare mature spores, and sterile water is added to prepare a suspension of slope spores. In the PDY medium shake flask, culture at 32℃, 150r / min for 48h, and put the spore suspension in the shake flask in a 500ML shake flask containing 100ML liquid medium at a ratio of 10%. The weight percentages of the optimized liquid fermentation medium components are as follows: 12% sucrose, 2% peptone, 0.1% yeast extract, K 2 HPO 4 0.1%, N-methyl-D-glucosamine 0.05%, NaCl0.5%, MgSO 4 0.02%, the rest is water, prepared by mixing uniformly, initial pH 6.02, cultured at 32°C, 180r / min for 60h. Centrifuge the fermentation broth at 5000r / min for 2 minutes, take the supernatant to test the enzyme activity, and use different inducer N-methyl-D-glucosamine dosage (weight p...

Example Embodiment

[0033] Example 3

[0034] Such as figure 1 As shown, a preparation method of transglucosidase, according to the production steps described in the content of the invention, aspergillus niger is cultivated on a slant for 7 days to prepare mature spores, and sterile water is added to prepare a slant spore suspension. In the PDY medium shake flask, culture at 32°C and 150r / min for 48h, and put the spore suspension in the shake flask in a 500ML shake flask containing 100ML liquid medium at a ratio of 10%. The weight percentages of the optimized liquid fermentation medium components are as follows: sucrose 5%, peptone 8%, yeast extract 0.5%, K 2 HPO 4 1%, N-methyl-D-glucosamine 0.05%, NaCl1%, MgSO 4 0.05%, the rest is water, prepared by mixing uniformly, initial pH 6.01, incubate at 32℃, 180r / min for 60h, centrifuge the fermentation broth at 5000r / min for 2 minutes, take the supernatant to test the enzyme activity, prepare The activity of the obtained crude enzyme solution was 20850u / M...

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Abstract

The invention relates to a preparation method of transglucosidase. The technical problem that during isomaltose hypgather production, the glucoside conversion rate is low is mainly solved. In order to solve the problem, according to the technical scheme, the aspergillus niger strain (General Microbiology Center of China Committee for Culture Collection of Microorganisms, CGMCC No.3.6478) serves as the original strain, a liquid fermentation culture medium is adopted, inductors are specially added, the aspergillus niger strain is induced to secrete a large amount of efficient transglucosidase through the four steps of slope cultivation, shake flask expanded cultivation, liquid deep fermentation and centrifugal separation, and then high-density transglucosidase fermentation liquid is prepared. The product can be applied to the key step, namely, the conversion stage, in isomaltose hypgather production.

Description

Technical field [0001] The invention relates to a preparation method of a transglucosidase, in particular to a preparation method of a transglucosidase with high transglycosidase ability, and belongs to the field of enzyme preparations and food additives. Background technique [0002] Glucosyltransferase (EC2.4.1.24), also known as α-transfer glucosidase, α-glucosidase, and in some cases also called α-D-glucosidase (α-D-glucosidase, EC3.2.1.20), α-glucosidase, maltase. Transglucosidase can cut the α-1,4 glycosidic bonds in the structure of sugar molecules such as maltose and oligomaltose, and transfer the free α-D-glucose residues to another α-D-glucose molecule or maltose molecule The α-1,6 position is the key enzyme preparation for the production of isomalt oligosaccharide. [0003] Isomaltooligosaccharide (IMO) is a kind of functional oligosaccharide with water-soluble dietary fiber function, low calorie, prevention of dental caries and other characteristics. It has become an ...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12R1/685
CPCC12N9/1051C12Y204/01024
Inventor 王方明谢永强徐保弟张宇
Owner SHANGHAI KINRY FOOD INGREDIENTS
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