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Polypeptide associated with the synthesis of 1-deoxynojirimycin, and a use therefor

A technology of deoxynojirimycin and polynucleotides, which is applied to polypeptides related to the synthesis of 1-deoxynojirimycin and its application fields, and can solve problems such as genes that are not isolated from polypeptides.

Active Publication Date: 2013-06-19
주휴온스푸디언스
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the polypeptide involved in the biosynthesis of 1-deoxynojirimycin is not known, and the gene encoding the polypeptide has not been isolated so far

Method used

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  • Polypeptide associated with the synthesis of 1-deoxynojirimycin, and a use therefor
  • Polypeptide associated with the synthesis of 1-deoxynojirimycin, and a use therefor
  • Polypeptide associated with the synthesis of 1-deoxynojirimycin, and a use therefor

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specific Embodiment approach

[0057] The present invention will be described in more detail below. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0058] The following examples are provided to illustrate preferred embodiments of the present invention, but are not intended to limit the scope of the present invention.

Embodiment 1

[0059] Example 1 Construction of a library using the genome of Bacillus subtilis MORI3K-85 producing 1-deoxynojirimycin

[0060] Strain culture

[0061] The improved Bacillus subtilis MORI3K-85 strain with improved α-glucosidase inhibitory activity was obtained from the isolated and identified Bacillus subtilis MORI (preservation number KCCM-10450) from Korean traditional food and irradiated with gamma rays. Specifically, the MORI strain was cultivated in Difco TMYM [Becton Dickinson and Company (BD), American BD Company] liquid medium, 180rpm shaking culture at 37°C for one day, and then irradiated with γ-rays (Soya GreenTec, South Korea) at a dose of 3kGy or 5kGy. The γ-ray-irradiated culture solution was spread on a YM agar plate and incubated at 37°C for 18 hours. Then, the formed colonies were transferred to fresh YM agar plates for incubation. Again, the colonies grown on the plate were inoculated into YM liquid medium, and cultured with shaking at 180 rpm at 37° C...

Embodiment 2

[0074] Embodiment 2 is screened by measuring inhibition of α-glucosidase activity

[0075] In order to select clones capable of producing 1-deoxynojirimycin from the white colonies, a method of measuring the activity of inhibiting α-glucosidase was used. The culture prepared in of Example 1 was heated at 100° C. for 10 minutes, and centrifuged at 8,000 rpm for 10 minutes. To 5 μL of the supernatant were added 75 μL of 0.1 M potassium phosphate buffer (pH 6.8), 50 μL of 12 mM p-nitrophenyl-α-glucopyranoside as a substrate, and 20 μL of crude enzyme solution. In order to prepare the crude enzyme solution, 0.8 g of rat small intestine acetone powder (Sigma, USA) was weighed and extracted in 100 mL of 0.1 M potassium phosphate buffer for 1 hr. The resulting solution was centrifuged at 12,000 rpm for 10 min, and the resulting supernatant was used as a crude enzyme solution.

[0076] The mixture was incubated at 37°C for 35 min, and the enzyme reaction was terminated with 50 μL o...

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Abstract

The present invention relates to a polypeptide associated with the biosynthesis of 1-deoxynojirimycin which is a substance that inhibits a-glucosidase activity, to a polynucleotide coding therefor, to a vector comprising the polynucleotide, to a transformant comprising the vector, and to a method in which the transformant is used in order to produce either a polypeptide associated with the synthesis of 1-deoxynojirimycin or 1-deoxynojirimycin. The polynucleotide of the present invention can be used in the volume production of 1-deoxynojirimycin, or can be used in studies searching for novel 1-deoxynojirimycin derivatives using recombinant technology, and hence is useful in developing pathogenic virus inhibitors and hypoglycaemic agents for diabetic patients.

Description

technical field [0001] The present invention relates to polypeptides related to the synthesis of 1-deoxynojirimycin and applications thereof, more specifically, the present invention relates to 1-deoxynojirimycin (which is an α-glucosidase inhibitor)-related to biosynthesis A polypeptide, a polynucleotide encoding the polypeptide, a vector containing the polynucleotide, a transformant containing the vector, and using the transformant to produce a polypeptide related to 1-deoxynojirimycin synthesis or 1-deoxynojirimycin Methods. Background technique [0002] With the Westernization of the living environment and eating habits and the increase of the elderly population, the onset of diabetes, a representative adult disease related to dietary life, has become more and more common in Korea. Diabetes mellitus, or simply diabetes, is a metabolic disease in which patients develop high blood sugar either because the pancreas does not produce enough insulin or because the cells of ea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/32C12N15/31C12N15/63C12P21/04
CPCC07K14/32C12N9/0004C12N9/1096C12N9/16C12N9/2405C12N15/52C12P17/12C12Y206/01019C12Y302/01084
Inventor 成洙一黄教列李载渊姜景敦朴永植赵勇锡陆媛晶
Owner 주휴온스푸디언스
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